2003 

ARS Immunology Research Workshop 

Proceedings 

A workshop sponsored by the U.S. Department of Agriculture, Agricultural 

Research Service at Bethesda, Maryland, on December 1-4, 2003. 

Also includes a bibliography of representative ARS immunology research published 

since 2000. 

Compiled and edited by: 

Tim Allen Rosemary Callahan 

United States Department of Agriculture United States Department of Agriculture 

Agricultural Research Service Agricultural Research Service 

National Agricultural Library National Program Staff 

10301 Baltimore Avenue 5601 Sunnyside Avenue 

Beltsville, Maryland 20705 Beltsville, Maryland 20705 

301/504-6212 

301/504-5175 

awic@nal.usda.gov

http://www.nal.usda.gov/awic

The use of trade, firm, or corporation names in this publication is for the 

information and convenience of the reader. Such use does not constitute an official 

endorsement or approval by the United States Department of Agriculture or the 

Agricultural Research Service of any product or service to the exclusion of others 

that may be suitable. 

2003 ARS Immunology Research Worskshop i 
Table of Contents 

EXECUTIVE SUMMARY.......................................................................................................................... 4 

WELCOME LETTER ................................................................................................................................. 7 

AGENDA....................................................................................................................................................... 8 

KEYNOTE ADDRESSES.......................................................................................................................... 14 

JAY A. BERZOFSKY ................................................................................................................. 14 

NATIONAL CANCER INSTITUTE, NIH, BETHESDA, MD ...................................... 14 

VACCINE STRATEGIES TO IMPROVE CTL AVIDITY, MEMORY, AND

EFFICACY..................................................................................................................... 14 

ARTHUR M. KRIEG .................................................................................................................. 16 

COLEY PHARMACEUTICAL GROUP, WELLESLEY, MA, USA. ........................... 16 

ENHANCING VACCINES WITH ADJUVANTS THAT ACTIVATE DENDRITIC

CELLS AND B CELLS IN VIVO ................................................................................ 16 

ALAN SHER ................................................................................................................................ 17 1

LABORATORY OF PARASITIC DISEASES, NIAID, NIH, BETHESDA, MD ........ 17 

THE P47 GTPASES: A FAMILY OF IFN INDUCED PROTEINS THAT 

REGULATE HOST RESISTANCE TO INTRACELLULAR PATHOGENS ........ 17 

PLENARY SESSIONS............................................................................................................................... 18 

INNATE IMMUNITY ................................................................................................................. 18 

IMMUNE REGULATION.......................................................................................................... 22 

IMMUNOGENETICS ................................................................................................................. 27 

IMMUNITY AND NUTRITION ................................................................................................ 30 

IMMUNITY AND DISEASE RESISTANCE............................................................................ 35 

IMMUNOINTERVENTION....................................................................................................... 39 

POSTER SESSIONS.................................................................................................................................. 43 

INNATE IMMUNITY ................................................................................................................. 43 

IMMUNE REGULATION.......................................................................................................... 47 

IMMUNOGENETICS ................................................................................................................. 53 

IMMUNITY AND THE PATHOGENESIS OF DISEASE...................................................... 55 

IMMUNITY AND NUTRITION ................................................................................................ 68 

IMMUNITY AND DISEASE RESISTANCE............................................................................ 75 

IMMUNOINTERVENTION STRATEGIES ............................................................................ 79 

WORKING GROUP REPORTS .............................................................................................................. 86 

IMMUNITY AND DISEASE RESISTANCE............................................................................ 87 

IMMUNOINTERVENTION....................................................................................................... 93 

IMMUNITY AND THE PATHOGENESIS OF DISEASES.................................................. 100 

NUTRITION AND IMMUNE FUNCTION INTERACTIONS............................................. 103 

BIBLIOGRAPHY OF ARS IMMUNOLOGY RESEARCH 2000-2003 ............................................. 108 

2003 ARS Immunology Research Worskshop 1 
AUBURN, ALABAMA ............................................................................................................................ 109

AQUATIC ANIMAL HEALTH RESEARCH ........................................................................ 109 

FAYETTEVILLE, ARKANSAS............................................................................................................. 115 

POULTRY PRODUCTION AND PRODUCTS SAFETY RESEARCH.............................. 115 

LITTLE ROCK, ARKANSAS ................................................................................................................ 119 

ARKANSAS CHILDREN'S NUTRITION CENTER............................................................. 119 

DAVIS, CALIFORNIA............................................................................................................................ 120

WESTERN HUMAN NUTRITION RESEARCH CENTER................................................. 120 

ATHENS, GEORGIA .............................................................................................................................. 124

SOUTHEAST POULTRY RESEARCH LABORATORY .................................................... 124 

WEST LAFAYETTE, INDIANA............................................................................................................ 131 

LIVESTOCK BEHAVIOR RESEARCH UNIT ..................................................................... 131 

WEST LAFAYETTE, INDIANA............................................................................................................ 132 

CROP PRODUCTION AND PEST CONTROL RESEARCH.............................................. 132 

AMES, IOWA........................................................................................................................................... 132 

NATIONAL ANIMAL DISEASE CENTER ........................................................................... 132 

NEW ORLEANS, LOUISIANA.............................................................................................................. 157 

SOUTHERN REGIONAL RESEARCH CENTER................................................................ 157 

BELTSVILLE, MARYLAND................................................................................................................. 159 

HENRY A. WALLACE BELTSVILLE AGRICULTURAL RESEARCH CENTER ........ 159 

BOSTON, MASSACHUSETTS .............................................................................................................. 180 

THE JEAN MAYER USDA HUMAN NUTRITION RESEARCH CENTER ON AGING 180 

EAST LANSING, MICHIGAN............................................................................................................... 182 

THE AVIAN DISEASE AND ONCOLOGY LABORATORY.............................................. 182 

MISSISSIPPI STATE, MISSISSIPPI..................................................................................................... 189 

STONEVILLE, MISSISSIPPI.................................................................................................. 189 

COLUMBIA, MISSOURI ....................................................................................................................... 190 

ANIMAL PHYSIOLOGY RESEARCH UNIT....................................................................... 190 

COLUMBIA, MISSOURI ....................................................................................................................... 192 

DONALD DANFORTH PLANT SCIENCE CENTER, PLANT GENETICS RESEARCH

UNIT............................................................................................................................................ 192 

CLAY CENTER, NEBRASKA............................................................................................................... 193 

ROMAN L. HRUSKA U.S. MEAT ANIMAL RESEARCH CENTER ................................ 193 

ORIENT POINT, NEW YORK .............................................................................................................. 193 

PLUM ISLAND ANIMAL DISEASE CENTER..................................................................... 193 

COLLEGE STATION, TEXAS.............................................................................................................. 197 

SOUTHERN PLAINS AGRICULTURAL RESEARCH CENTER...................................... 197 

2003 ARS Immunology Research Worskshop 2 
HOUSTON, TEXAS................................................................................................................................. 201 

CHILDREN'S NUTRITION RESEARCH CENTER ............................................................ 201 

LOGAN, UTAH........................................................................................................................................ 202 

POISONOUS PLANT RESEARCH......................................................................................... 202 

PULLMAN, WASHINGTON.................................................................................................................. 204 

ANIMAL DISEASE RESEARCH UNIT................................................................................. 204 

KEARNEYSVILLE, WEST VIRGINIA................................................................................................ 208 

NATIONAL CENTER FOR COOL AND COLD WATER AQUACULTURE .................. 208 

LARAMIE, WYOMING ......................................................................................................................... 209 

ARTHROPOD-BORNE ANIMAL DISEASES RESEARCH LABORATORY.................. 209 

APPENDIX ............................................................................................................................................... 211 

LIST OF PARTICIPATNTS................................................................................................................... 212 

AUTHOR INDEX..................................................................................................................................... 226 

2003 ARS Immunology Research Worskshop 3 
2003 ARS Immunology 

Research Workshop 

Executive Summary 

The 2003 ARS Immunology Research Workshop was sponsored by the United States 

Department of Agriculture (USDA), Agricultural Research Service (ARS), in Bethesda, 

Maryland, December 1-4, 2003. The purpose of the workshop was to exchange recent 

scientific breakthroughs in animal immunology and determine where ARS and its 

partners should concentrate future efforts to significantly advance national research 

programs in animal health, animal production, aquaculture, food safety, and human 

nutrition. 

The workshop participants included ARS immunologists working in livestock, wildlife, 

avian and marine species, as well as immunologists working in other Federal laboratories 

and universities across the United States. The first day of the workshop provided a series 

of presentations covering high profile topics in basic immunology research with emphasis 

on cutting-edge discoveries in innate immunology, immune regulation, and 

immunogenetics. The second day focused on the applications of immunology research in 

nutrition, disease resistance, and immunointervention. This exchange of information set 

the stage for the third day, when all participants were divided into the following four 

working groups: 1) Immunity and disease resistance; 2) Immunointervention strategies; 

3) Immunity and the pathogenesis of diseases; 4) Immunity and nutrition. The challenge 

set forth for the working groups was to identify research objectives within specific areas 

of animal immunology that could significantly advance research programs in animal 

health, animal production, aquaculture, food safety, and human nutrition.

The goal for the pathogenesis working group was to provide research priorities to 

advance our understanding of immune responses that lead to pathogenesis, either 

inherently programmed within the immune system, or driven by metabolic and infectious 

disease pathways. The human nutrition working group was asked to identify research 

objectives in animal immunology that were most likely to make significant contributions 

to our understanding of health and nutrition. The disease resistance working group was 

asked to identify research objectives that will lead to the discovery and characterization 

of immune mechanisms associated with heritable disease resistant traits and that may in 

parallel be used as targets to design novel biotherapeutics and highly efficacious 

vaccines. The immunointervention working group was tasked with providing research 

2003 ARS Immunology Research Worskshop 4 
objectives that will significantly increase our understanding of protective immunity and 

lead to the rational design of highly efficacious vaccines and diagnostics.

A significant outcome of the workshop was that all four working groups identified the 

lack of veterinary immunological reagents as a key stumbling block to advancing the 

field of animal immunology. The lack of immunological reagents was also considered 

paramount to make significant progress in many of our national research programs. For 

instance, the lack of efficient tools for evaluating cell-mediated immunity in cattle and 

swine makes the discovery and development of highly efficacious vaccines more difficult 

to attain and demonstrate. Many of the workshop participants felt that cell-mediated 

immunity should be at the core of strategies to discover new tools against the diseases 

that resist traditional vaccine approaches. The overwhelming consensus was that the void 

in our immunological tool chest is the rate-limiting step in providing the next generation 

of vaccines.

In today's economy it is highly unlikely that the private sector will invest in the basic 

research necessary to develop the tools for testing cell-mediated immunity. Accordingly, 

26 participants agreed to look at organizing a community wide effort, post-workshop, to 

discover, develop, and distribute quality-controlled immunological reagents for 

veterinary science research. 

As a result of the final working sessions, ARS immunologists and their partners identified 

a total of 66 research objectives to incorporate into future research action plans. The 

research objectives with the highest priority are noted here: 

• Establish national and international networks to prioritize and develop immune 

toolkits for animal species of economic importance. Coordinate tool development so 

that priority reagents become available. Work with commercial sources to market 

products. 

• Identify critical components of innate immunity; Use innate immune pathways for 

interventions/traditional and therapeutic vaccines and for development of new 

adjuvants and improvement of vaccine efficacy. 

• Identify and characterize host genes that control important immunologic 

responses; determine whether alleles of certain genes would enhance animal 

health 

• Use information from microbial genomics to define substances from pathogens 

that stimulate/regulate immune responses at mucosal surfaces. 

• Develop broad spectrum, targeted delivery systems (target dendritic cells or

others) for immunomodulator and/or peptide and/or DNA delivery to effector 

cells. 

• Optimize use of existing vaccines and immunomodulators for intervention at 

embryogenesis, birth, weaning, or prior to stress (shipping, comingling, 

slaughter). 

• Elucidate factors affecting efficacy of current adjuvant systems 

• Define the affects of stress/diet/age on immune responses at mucosal surfaces. 

• Identify specific nutrients with beneficial immune modulating properties that 

optimize immune function. Define specific nutrient requirements for effective and 

2003 ARS Immunology Research Worskshop 5 
appropriate vaccination and immunological memory, and for amelioration of 

periods of immune deficiency, such as during pregnancy, parturition, pediatric 

development, ageing, and stress. 

• Define probiotic microbial populations that affect immune development and 

functional resistance at mucosal surfaces and on the efficacy of systemic 

immunity, and the diets (prebiotics) that support appropriate resident or ingested 

microbial populations. 

• Develop new methods for detecting and measuring cell-mediated immunity; 

identify microbial antigens and regulatory events critical for the development and 

maintenance of cell-mediated immunity. 

• Utilize genomic information to reveal new approaches for disease control or 

biotherapeutic production and identify novel nutrient and immune interactions. 

• Define gene expression patterns and regulatory phenomena that affect the level of 

host resistance at mucosal surfaces. 

In addition, the workshop participants selected the following research objectives for their 

potential to substantially impact ARS national research programs in animal health, animal 

production, aquaculture, food safety, and human nutrition. 

• Novel approaches for disease diagnostics; Biomarkers of immunointervention 

success will enhance our capacity to screen for the most efficacious method to 

elicit a protective immune response. 

• Improved methods to limit immune-associated disease and persistence of 

pathogens thereby increasing production and decrease spread of disease, 

respectively.

• Improved methods to limit factors (e.g., environmental, husbandry, nutritional) 

that negatively impact neonatal immune development. 

• Definition of pathways for interventions, for development of both traditional and 

therapeutic vaccines, and for preparation of novel mucosally targeted 

biotherapeutics 

• Increased understanding of how innate immunity impacts vaccine responses; 

improved knowledge base for adjuvant and immune modulator design/selection 

for vaccines and biotherapeutics. 

• Improved vaccine efficacy; verification of efficacy of mucosal delivery vectors 

and adjuvants; targeting the neonate and maternal immunity. 

• Broad-based vaccine delivery systems applicable to both production animals and 

wildlife that account for the practical realities of immunization in the field. 

• Provide identification of diets that enhance immune development and reduce 

inflammation. 

• Better understanding of the interaction between dietary choices and environment 

on immune function and optimal health. 

• The emergence of genetic profiles will include dietary recommendations for 

human life style and animal management to optimize health. 

• By leveraging knowledge in databases from human disease research, we will 

build a comparative immunology database that can be a guide to antigen selection 

for use in vaccines. 

2003 ARS Immunology Research Worskshop 6 
Welcome Letter 

United States Department of Agriculture 

Agricultural Research Service 

National Program Staff 

December 1, 2003 

Dear Workshop Attendee: 

On behalf of the National Program Staff, welcome to the “2003 ARS Immunology Research 

Workshop.” We are delighted that you could join us for this exiting event. The scientists 

participating in the workshop include Immunologists working in very different fields and we 

hope the diversity of disease models, animal species, and approaches to solving problems 

discussed over the next three days will be beneficial to your research. We also hope you will have 

many opportunities to establish strategic collaborations over the next three days that will build on 

our understanding of Immunology and its many applications to animal and human health. 

We have four goals for the workshop: 

1. To exchange new information and ideas in immunology to maximize the impact of our 

research programs and solve problems of high National priority. 

2. To enhance interactions and form strategic collaborations between scientists engaged in 

immunology research. 

3. To provide a forum for stakeholders, customers, and partners to give input and direction on 

current and future ARS immunology research programs. 

4. To initiate the preparation of action plans for consideration at future ARS National Program

workshops. 

The workshop has been structured to facilitate interactions and the exchange of new ideas 

between scientists conducting research in Federal laboratories, academia, and the private sector. 

Working group sessions will be held on the last day to assess future research direction and 

prepare research plans. We are very fortunate to have some of the most prominent experts in 

immunology working in animal and human health participate in this workshop. We have tried to 

format this workshop to promote constructive exchange of the most recent data and new ideas. 

Once again, we are delighted that you could join us for this exciting event and we thank you for 

your participation in this workshop. 

Sincerely,

Cyril Gerard Gay, D.V.M., Ph.D 

National Program Leader, Animal Health and Safety

Animal Production, Product Value, and Safety

5601 Sunnyside Avenue $ Beltsville, MD 20705-5138 

2003 ARS Immunology Research Worskshop 7 
2003 ARS Immunology Research Workshop 

Agenda 

December 1-4, 2003 

Bethesda, Maryland 

December 1, 2003 

Kick Off 

18:00 – Welcome Address, Cyril Gerard Gay, National Program Leader, Animal Health 

and Safety, ARS, Beltsville, MD 

Goals, partnerships, and opportunities 

18:30 - Keynote Presentation, Jay Berzofsky, NCI/NIH/HHS, Bethesda, MD 

Vaccine strategies to maximize CTL avidity, memory, and efficacy 

19:30 - Reception 

December 2, 2003 

Plenary Sessions 

Innate Immunity

Moderator: James A. Roth, Director, Center for Food Security and Public Health, 

College of Veterinary Medicine, Iowa State University, Ames, IA 

08:00 - Matthew Fenton, University of Maryland School of Medicine, Baltimore, MD 

Role of Toll-like Receptors in the Host Response Against Mycobacterial Infection 

08:30 - Susan D. Eicher, ARS, Livestock Behavior Research Unit, West Lafayette, IN 

Toll-like Receptor and Acute Phase Cytokine Expression in Neonatal Dairy Calves 

09:00 - Rami Dalloul, ARS, Animal Parasite Diseases Laboratory, Beltsville, MD 

Oligodeoxynucleotides (ODN)-induced activation of macrophage-mediated innate 

immunity against intracellular pathogens, Eimeria and Salmonella, and the 

development of novel immunomodulatory strategies using embryo injection of CpGODNs in poultry. 

2003 ARS Immunology Research Worskshop 8 
09:30 - Elida M. Bautista, ARS, Foot and Mouth Disease Research, Orient Point, NY 

Analysis of swine skin-derived dendritic cells in the induction of innate immune 

responses to Foot-and-mouth disease virus (FMDV) 

10:00 - Interactive Poster Session (38 posters) 

• Innate Immunity 

• Immune Regulation 

• Immunogenetics 

• Immunity and the pathogenesis of diseases 

Immune Regulation

Moderator: William Golde, Immunologist, Plum Island Animal Research Center, ARS, 

Orient Point, NY 

11:00 - Willi Born, National Jewish Medical and Research Center, Denver, CO 

Regulatory role of gamma/delta T cells in allergic airway hypereactivity 

11:30 - David Woodland, Trudeau Institute, Saranac Lake, NY 

Regulation of Memory T cell Responses in Viral Infections of the Respiratory Tract 

12:00 - Lunch and Keynote Presentation: Arthur Kreig, Coley Pharmaceuticals, Wellesley, 

MA 

Enhancing vaccines with adjuvants that activate dendritic cells and B cells in vivo 

13:30 - Wendy Brown, Washington State University, Pullman, WA

Regulation of the CD4+ T lymphocyte response by Anaplasma marginale 

14:00 - Wenbin Tuo, ARS, Animal Parasite Diseases Laboratory, Beltsville, MD 

Bovine T cell response to antigens of Neospora caninum 

14:30 - Marc Estes, University of Texas, Galveston, TX 

Progress towards developing effective subunit vaccine responses by CD8+ memory and 

effector T cells in a bovine model of tuberculosis 

15:00 – Jesse P. Goff, ARS, Periparturient Diseases of Cattle Research Unit, Ames, IA 

Effects of the Mammary Gland on the Composition and Function of Peripheral Blood 

Mononuclear Leukocyte Populations in Periparturient Dairy Cows

15:30 - Break 

Immunogenetics

Moderator: Peter L. Nara, Director of Research and Development, Biological Mimetics, 

Inc., Frederick, MD 

2003 ARS Immunology Research Worskshop 9 
16:00 - Annie DeGroot, Epivax, Inc, Providence, RI 

Accelerating vaccine design: new in silico epitope mapping methods 

16:30 - Henry Hunt, ARS, Avian Diseases and Oncology Laboratory, East Lansing, MI 

Marek's disease and the MHC: How viruses subvert the Immune System 

for their advantage 

17:00 - Lynn M. Herrmann, ARS, Animal Disease Research Unit, Pullman, WA

Expression of specific MHC Class II DRB1 alleles associates with sheep persistently 

infected with ovine progressive pneumonia virus (OPPV) 

17:30 - John Butler, University of Iowa, IA 

From Basic Porcine Immunology to Practical Application 

December 3, 2003 

Plenary Sessions 

Immunity and Nutrition

Moderator: Ildy Katona, Department of Pediatrics, Uniformed Services University of

the Health Sciences in Bethesda, MD

07:30 - Joe Urban, ARS, Nutrient Requirements and Functions Laboratory, Beltsville, 

MD 

The effect of human-derived probiotic bacteria on the immune and intestinal function 

of pigs

08:00 - Kirk Klasing, University of California, Davis, CA 

The care and feeding of an immune system: How much is needed and what are the 

priorities?

08:30 - Simin Meydani, ARS, Nutritional Immunology Laboratory, Boston, MA 

Aging, immune response, and infectious disease: molecular mechanisms and reversal 

by nutrient intervention

09:00 - Brian Nonnecke, Periparturient Diseases of Cattle Research Unit, Ames, IA 

Nutritional Plane Affects Antigen-Induced Proliferation of Lymphocyte Subsets in 

Milk Replacer-Fed Calves Vaccinated with M. bovis Bacillus Calmette-Guerin (BCG)

09:30 - Harry Dawson, Nutrient Requirements and Functions Laboratory, Beltsville, MD 

Real-Time PCR Arrays Can Delineate Immunological and Nutrition-Related Gene 

Expression in Swine 

2003 ARS Immunology Research Worskshop 10 
10:00 - Interactive Poster Session (32 posters) 

• Immunity and nutrition 

• Immunity and disease resistance 

• Immunointervention strategies 

Immunity and Disease Resistance

Moderator: Muquarrab Qureshi, National Program Leader, Animal Genomics, 

CSREES, USDA, Washington D.C. 

11:00 – Michael Murtaugh, University of Minnesota, 

Mechanisms of Innate and Adaptive Immunity in Swine 

11:30 - Lou Gasbarre, ARS, Bovine Functional Genomics, Beltsville, MD 

Genetic and Immunologic basis for resistance to GI nematode infections 

12:00 - Lunch and Keynote Presentation: Alan Sher, NIAID/NIH/HHS, Bethesda, MD 

The p47 GTPases: a family of IFN induced proteins that regulate host resistance to 

intracellular pathogens 

13:30 - Pamela Ferro, ARS, Food and Feed Safety Research, College Station, TX 

Heterophils Isolated From Chickens Resistant to Extraintestinal Salmonella enteritidis 

Infection Express Higher Levels of Pro-inflammatory Cytokine mRNA Following 

Infection Than Heterophils From Susceptible Chickens

14:00 - Hyun Lillehoj, ARS, Animal Parasite Diseases Laboratory, Beltsville, MD 

High-throughput molecular approaches to identify coccidiosis disease resistance genes 

and protective immunological mechanisms in avian coccidiosis 

Immunointervention

Moderator: Ronald Schultz, Professor and Chair, Department of Pathobiological 

Sciences, University of Wisconsin-Madison, WI 

14:30 – Cynthia Baldwin, University of Massachussets, 

Protective Killed Leptospira Vaccine Induces Potent Type 1 (Th1) Immunity 

Comprising Responses by CD4 and WC1+ γδ T Lymphocytes 

15:00 – Evelyn Dean-Nystrom, ARS, Food Safety and Enteric Diseases Research Unit, 

Ames, IA 

Development of an Oral Vaccine for Escherichia coli O157:H7 

15:30 - Break 

2003 ARS Immunology Research Worskshop 11 
16:00 - Wongi Min, ARS, Animal Parasite Diseases Laboratory, Beltsville, MD 

Adjuvant Effect of IL-1β, IL-2, IL-8, IL-15, IFN-α, IFN-γ, TGF-β4 and lymphotactin 

on DNA vaccination against Eimeria acervulina

16:30 - Haiqi He, ARS, Food and Feed Safety Research, College Station, TX 

Stimulation of chicken leukocytes and reduction of Salmonella organ invasion in 

neonatal chicken by immunostimulatory CpG-oligodeoxynucleotide 

17:00 – M. Suresh, Assistant Professor of Immunology, University of Wisconsin, 

Madison, WI 

Regulation of Memory CD8 T Cell Generation by Cytokines 

December 4, 2003 

Research Priorities Working Groups. The workshop participants will be divided into 

teams and asked to identify research priorities, strategic collaborations, and innovative 

research plans that will substantially increase our knowledge of immunology and deliver 

solutions to disease problems of high national priority in animal and human health. The 

recommendations of the participants will form the basis for developing ARS National 

Program action plans. Focus will be given to the following research areas: 

• Immunity and disease resistance 

• Immunointervention strategies 

• Immunity and the pathogenesis of diseases 

• Nutrition and immune function interactions 

08:30 Working Group Instructions

Robert Heckert, National Program Leader, Animal Health, ARS, Beltsville, MD 

Lewis Smith, National Program Leader, Aquaculture & Animal Well-Being, 

ARS, Beltsville, MD 

09:00 Working Group Sessions 

• Immunity and Disease Resistance 

Working Group Leaders: 

3⁄4 Lanie Bilodeau, Immunologists, Catfish Genetics Research Unit, ARS, 

Stoneville, MS 

3⁄4 Lou Gasbarre, Research Leader, Bovine Functional Genomics Research 

Unit, Animal and Natural Resources Institute, ARS, Beltsville, MD 

3⁄4 Michael H. Kogut, Immunologist, Southern Plains Agricultural Research 

Center, ARS, College Station, TX 

3⁄4 Joan Lunney, Immunologist, Animal Parasite Diseases Laboratory, 

Animal and Natural Resources Institute, ARS, Beltsville, MD 

2003 ARS Immunology Research Worskshop 12 
Immunointervention Strategies 

Working Group Leaders: 

3⁄4 Marcus Kehrli, Research Leader, Virus and Prion Diseases Research 

Laboratory, National Animal Disease Center, ARS, Ames, IA

3⁄4 Hyun Lillehoy, Immunologist, Animal Parasite Diseases Laboratory, 

Animal and Natural Resources Institute, ARS, Beltsville, MD 

3⁄4 David White, Veterinary Medical Officer, Arthropod-borne Animal 

Diseases Research Laboratory, ARS, Laramie, WY 

• Immunity and the Pathogenesis of Diseases 

Working Group Leaders: 

3⁄4 Don Knowles, Research Leader, Animal Disease Research Unit, ARS, 

3⁄4 Pullman, WA 

3⁄4 David Swayne, Research Leader, Southeast Poultry Research Laboratory, 

ARS, Athens, GA 

3⁄4 Ray Waters, Veterinary Medical Officer, Bacterial Diseases of Livestock 

Research Unit, National Animal Disease Center, ARS, Ames, IA 

• Nutrition and Immune Function Interactions 

Working Group Leaders: 

3⁄4 Theodore Elsasser, Research Animal Scientist, Growth Biology 

Laboratory, Animal and Natural Resources Institute, ARS, Beltsville, MD 

3⁄4 Simin Meydani, Chief, Nutritional Immunology Laboratory, Jean Mayer 

USDA Human Nutrition Research Center on Aging, ARS, Tufts University, 

Boston, MA 

3⁄4 Joe Urban, Research Leader, Nutrient Requirements and Functions 

Laboratory, Beltsville Human Nutrition Research Center, ARS, 

 Beltsville, MD 

12:00 Lunch 

Working Group Reports 

13:00 - Immunity and Disease Resistance 

13:30 - Immunointervention Strategies 

14:00 - Immunity and the Pathogenesis of Diseases 

14:30 - Nutrition and Immune Function Interaction 

15:00 – Cyril Gerard Gay, National Program Leader, Animal Health and Safety, ARS, 

Beltsville, MD 

Conclusion and post-meeting instructions 

2003 ARS Immunology Research Worskshop 13 
Keynote Addresses 

December 1, 2003

Jay A. Berzofsky 

Molecular Immunogenetics & Vaccine Research Section, Metabolism Branch, 

National Cancer Institute, NIH, Bethesda, MD 

Vaccine strategies to improve CTL avidity, 

memory, and efficacy 

Cytokines are potentially useful not only as vaccine components to increase the 

magnitude of the immune response, but also to steer it in desired directions. We found 

that certain combinations such as GM-CSF and IL-12 synergized to increase cytotoxic T 

lymphocyte (CTL) activity. We showed that GM-CSF worked to increase antigen 

presenting cell numbers and activity in the draining lymph nodes. We reasoned that 

CD40L might synergize with GM-CSF by maturing those presenting cells. Indeed, GMCSF and CD40L were a potent combination to increase CTL responses. Also, we had 

seen that NKT cells making IL-13 dampened the CTL response to tumors, so we asked if 

blockade of IL-13 or elimination of NKT cells would also increase the CTL response to a 

peptide vaccine. We found that either of these treatments further enhanced the CTL 

response and the ability to protect against viral challenge. Thus, a push-pull approach in 

which a cytokine and a costimulatory molecule are combined to push the response and 

disruption of an inhibitory pathway is used to pull the response can maximize CTL 

induction. 

In addition to increasing the quantity of the CTL, we sought to also improve the quality

of the CTL induced by a vaccine. We previously found that avidity of CTL is critical for 

effective viral clearance, but selective induction of high avidity CTL has been difficult to 

achieve with vaccines. Likewise, persistent CTL memory is essential for long-lasting 

CTL-mediated vaccine efficacy. Thus, we have sought strategies to selectively induce 

longer-lived high avidity CTL with vaccines targeting HIV protein antigens. First, 

increasing costimulation (signal 2) allowed us to immunize with lower doses of antigen

(signal 1) to selectively induce high avidity CTL in vivo. This was achieved using a 

recombinant poxvirus vector expressing a triad of costimulatory molecules (B7-1, ICAM1, and LFA-3) (TRICOM) to increase expression of these molecules on splenocytes 

pulsed with an HIV-1 CTL epitope peptide. Second, since IL-15 was known to 

contribute to maintenance of CTL memory, we asked whether IL-15 at the time of 

immunization would alter the CTL response. Expression of IL-15 by a recombinant 

vaccinia vector along with HIV-1 gp160 induced long-lived memory CTL that persisted 

at least 14 months after a single boost. The memory cells expressed higher levels of IL2003 ARS Immunology Research Worskshop 14 
15Rα and showed greater proliferation to IL-15 in vitro and homeostatic proliferation in 

vivo, as well as higher avidity for antigen. Thus, IL-15 at the time of priming selects or 

induces a different phenotype CTL with greater avidity and longevity. These vaccine 

strategies can provide more effective CTL immunity by enhancing the numbers, avidity, 

and memory of the CTL induced, allowing for greater and longer lasting protection. 

2003 ARS Immunology Research Worskshop 15 
December 2, 2003

Arthur M. Krieg 

Coley Pharmaceutical Group, Wellesley, MA, USA. 

Enhancing vaccines with adjuvants that activate 

dendritic cells and B cells in vivo 

The innate immune system has evolved “pattern recognition receptors” that detect 

molecules present in various classes of pathogens, but not in our own cells. The best 

known of these pattern recognition receptors belong to the family of Toll-like receptors 

(TLRs), which appear to have evolved to activate appropriate protective immune 

responses when a pathogen is detected. One such pathogen-associated pattern is 

unmethylated CpG dinucleotides in bacterial or viral DNA, which are detected by TLR9, 

thereby stimulating innate and adaptive immunity. These immune effects can be 

mimicked by synthetic oligodeoxynucleotides containing CpG motifs (CpG ODN). In 

humans and most other species, at least two types of immune cells express TLR9 and are 

activated directly by CpG ODN; B cells, and plasmacytoid dendritic cells (pDC). CpG 

ODN activate B cells to proliferate and secrete immunoglobulin; and pDC to secrete a 

variety of Th1-like cytokines, chemokines, and type I interferons, and to express 

increased costimulatory molecules. When activated by CpG, pDC gain the ability to 

stimulate Th1-like T cell responses. CpG ODN induce immune defense mechanisms that 

protect against challenge with a wide range of infectious pathogens, and have shown 

therapeutic activity in animal models of allergic disease and cancer. CpG ODN are also 

extremely effective vaccine adjuvants, inducing Th1 responses in mice, primates, and 

other species with both specific antibody and CTL. Phase I human clinical trial results 

indicate that a CpG ODN, 7909, is relatively well tolerated. As an adjuvant for a 

hepatitis B vaccine, CpG 7909 appears to induce earlier seroconversion with the 

production of increased levels of specific antibody and cellular responses. A phase II 

human clinical trial of CpG 7909 in HIV-infected subjects, including prior hepatitis B 

vaccine nonresponders, showed a striking increase in the magnitude of antibody 

responses in the subjects receiving CpG as a vaccine adjuvant. CpG ODN are produced 

by cGMP chemical synthesis, have excellent stability and solubility, and show an 

excellent safety profile, suggesting wide utility as vaccine adjuvants for veterinary 

applications.

2003 ARS Immunology Research Worskshop 16 
December 3, 2003

Alan Sher 

1

Laboratory of Parasitic Diseases, NIAID, NIH, Bethesda, MD 

2

Center for Biologics Evaluation and Research, FDA, Bethesda, MD 

3

Dept. of Medicine and Immunology, Duke University Medical Center, Durham, NC 

The p47 GTPases: a family of IFN induced 

proteins that regulate host resistance to 

intracellular pathogens 

Alan Sher1, C. Feng2, C. Collazo1, and G.A. Taylor3

Activation of the immune system by interferon-γ is critical for host resistance to many 

infectious agents. This lymphokine induces the expression of a wide range of effectors 

that undermine the ability of pathogens to survive in host cells. Among these is a newly 

discovered family of 47 kDa GTPases that provide resistance to intracellular protozoa, 

bacteria, and viruses. Elimination of different p47 GTPases in mice through gene 

targeting severely cripples IFN-γ-regulated defense against Toxoplasma gondii, Listeria

monocytogenes, Mycobacterium sp., and other pathogens. Interestingly, different 

members of this gene family appear to effect resistance to different groups of pathogens. 

Current evidence suggests that p47 GTPases function against pathogens at two different 

levels: by facilitating intracellular killing of pathogens and by promoting survival of 

effector T lymphocytes. 

2003 ARS Immunology Research Worskshop 17 
Plenary Sessions 

December 2, 2003

Innate Immunity 

Role of toll-like receptors in the host response against mycobacterial infection

Matthew J. Fenton 

Director of Research, Division of Pulmonary and Critical Care Medicine; Professor of 

Medicine, Microbiology and Immunology, University of Maryland School of Medicine, 

Baltimore, MD 

Toll-like receptor (TLR) proteins mediate cellular activation by microbes and microbial 

products. To delineate the role of TLR proteins in the development of host immune 

responses against mycobacteria, wild-type and TLR-deficient mice were infected with 

non-pathogenic Mycobacterium bovis BCG (BCG). Two weeks after intraperitoneal 

challenge with BCG, few bacilli were present in the lungs of wild-type and TLR4-/- 

mice, whereas bacterial loads were 10-fold higher in the lungs of infected TLR2-/- mice. 

BCG challenge in vitro strongly induced pro-inflammatory cytokine secretion by 

macrophages from wild-type and TLR4-/- mice, but not by TLR2-/- macrophages. In 

contrast, intracellular uptake, intracellular bacterial growth, and suppression of 

intracellular bacterial growth in vitro by IFN-g were similar in macrophages from all 

three mouse strains, suggesting that BCG growth in the lungs of TLR2-/- mice was a 

consequence of defective adaptive immunity. Antigenic stimulation of splenocytes from 

infected wild-type and TLR4-/- mice induced T cell proliferation in vitro, whereas T cells 

from TLR2-/- mice failed to proliferate. This lack of T cell proliferation in TLR2-/- mice 

correlated with an inability of CD11c+ dendritic cells to be activated in vivo following 

BCG challenge. Unexpectedly, activated CD4+ T cells from both TLR-deficient mouse 

strains secreted little IFN-g in vitro, compared with control T cells. A role for TLR4 in 

the control of bacterial growth and IFN-g production in vivo was observed only when 

mice were infected with higher numbers of BCG. Thus, TLR2 and TLR4 appear to 

regulate distinct aspects of the host immune response against BCG. 

Toll-like receptor and acute phase cytokine expression in neonatal dairy 

calves

S. D. Eicher1

, T. A. Johnson2, K. A. McMunn1, and T. R. Johnson2 

1

 USDA-ARS, Livestock Behavior Research Unit, West Lafayette, IN and 2Purdue 

University, West Lafayette, IN 

The well-being of farm animals is dependent on many factors, so that a multi-disciplinary 

approach is essential. The Livestock Behavior Research Unit uses behavior, physiology, 

and immunology measures to answer well-being questions for poultry, swine, and dairy 

2003 ARS Immunology Research Worskshop 18 
cattle. The innate immune system is frequently the first immune responder during many 

stressors and acute phase cytokines affect behavior and learning. Toll-like receptors are 

the pathogen recognition molecules that initiate the cascade that increases the acute phase 

cytokines. The effect of stressors on the toll-like receptors of farm animals is not known. 

Studies have been completed that examine the effect of stimulants on RNA expression of 

toll-like receptors in young dairy calves. The objective of study one was to assess the 

optimum concentration and times of incubation for expression of TLR2 and TLR4 after 

in-vitro stimulation of bovine monocyte derived macrophages with Saccharomyces 

cerevisiae (yeast cell-wall) beta-glucan (BG), Escherichia coli lipopolysaccharide (LPS), 

and Staphylococcus aureus peptidoglycan (PGN). Monocyte derived macrophages from 

10 donor calves (5 to 6 wk-of-age) were first stimulated with LPS or PGN at 1, 5, or 

25μg/ml or BG at 20, 40, or 80μg/ml for one hour at 37°C. Then the optimum 

concentrations were used to incubate cells at 30, 60, or 90 min. Real-time RT-PCR was 

used to quantify RNA expression of TLR2 and TLR4. RNA from monocyte derived 

macrophage expressed more bovine TLR4 than TLR2. Sixty minutes appears to be an 

adequate time of stimulation to change TLR4 expression, but at least 90 minutes are 

needed to attain maximum changes in TLR2 expression. The objective of study two was 

to determine if in vitro stimulation of whole blood leukocytes with beta-glucan plus 

ascorbic acid (a potential immunomodulator) was dependent on the age of the calf. 

Blood samples were taken from 12 non-transported Holstein calves at 1, 3, 7, 10, 14, 18, 

21, 24, and 28 d-of-age. Leukocytes were stimulated with ascorbic acid (0.3 ug/ml) plus 

beta-glucan (0.4 ug/ml) or media only for 1 h and then RNA was extracted. The RNA 

was subjected to real-time RT-PCR for quantification of the expression of interleukin-1 

(IL-1) and its receptor antagonist (IL-1Ra), and toll-like receptors 2 and 4 (TLR2 and 

TLR4). TLR2 and TLR4 had treatment effects (P<.05), but not day or treatment by day 

interactions. TLR2 was greater (P<.05) for treated cells on day 7, 14, and 24 and tended 

(P<.10) to be greater on day 10. In contrast, TLR4 was only greater for treated cells on 

day 7 (P<.05) and tended to be (P<.10) on day 24. IL-1 had a treatment main effect and a 

treatment by day interaction (P<.05), but IL-1Ra had main effects for treatment and day 

(P<.05), but only a trend (P<.10) for a treatment by day interaction. IL-1 was greater for 

treated cells (P<.05) on all but day 4. IL-1Ra was greater (P<.05) for treated cells only on 

days 1, 7, 10, and 24. Only IL-1 and its receptor antagonist expression were stimulated on 

day 1. On days 7 and 24 all tested receptors and cytokines had increased RNA 

expression. So, it appears that there are periods during which the blood leukocytes may 

be refractory for increased RNA expression of cytokines and toll-like receptors in 

response to beta-glucan and ascorbic acid stimulus. These data enhance our 

understanding of the biological kinetics of the developing neonatal calf immune system, 

so that management procedures can occur at the best possible times. 

2003 ARS Immunology Research Worskshop 19 
Oligodeoxynucleotides (ODN)-induced activation of macrophage-mediated 

innate immunity against intracellular pathogens, Eimeria and Salmonella, 

and the development of novel immunomodulatory strategies using embryo 

injection of CpG-ODNs in poultry. Dalloul1,2

Rami, Lillehoj1 Hyun S., Xie2 Hang, 

Heckert2 Robert A., Babu3 Uma, Raybourne3 Richard, and Klinman4 Dennis M. 

1

 USDA-ARS, Animal Parasitic Diseases Laboratory, ANRI,, Beltsville, MD, 20705, 

2

VA-MD Regional College of Veterinary Medicine, University of Maryland, College 

Park, MD, 20742, 3

Center for Food Safety & Applied Nutrition, US FDA, Laurel, MD 

20708, 4

Section of Retroviral Immunology, Center for Biologics Evaluation & Research, 

US FDA, Bethesda, MD, 20892. 

The immunostimulatory properties of synthetic CpG oligodeoxynucleotides (ODNs) have 

been well studied in various mammalian models including humans and mice. However, 

little was known about effects of CpG-ODNs on immune responses of chickens, a 

common avian species with important economical value in the poultry industry. ODN 

2006 which has been reported to be an optimal stimulatory sequence for humans, showed 

strong immunomodulatory effects on HD11 cells. Whereas ODN 1826, a murine optimal 

CpG-ODN had weak influences on HD11 cells. We also observed that 2006 induced 

strong IL-6 and nitric oxide secretion by HD11 cells in both dose- and time-dependent 

manners. 2006 induced activation also resulted in increased intracellular killing of S. 

enteritidis (SE) by HD11 cells. Furthermore, 2006 activated HD11 cells had reduced 

proliferation and underwent apoptosis after CpG stimulation. Further studies were carried 

out to test several short synthetic CpG ODNs for their immunomodulatory effects on 

disease susceptibility in Eimeria acervulina. On day 18 of incubation, specific pathogenfree embryos were injected (air cell) with either 25 μg or 50 μg of CpG ODN (CpG-1, 

CpG-2, CpG-3 and CpG-4), a non CpG-containing control (CpG-0), or a negative control 

(PBS, phosphate buffered saline). At one week of age, birds were orally inoculated with 

104 E. acervulina oocysts. Body weights and body weight gains were not affected by any 

of the treatments. However, among CpG ODN treatments, a differential antibody 

response was observed with the lowest response in CpG-2 birds and highest in CpG-4 

group. This study demonstrates that CpG ODN were effective immunoprotective agents 

in chickens, and the potentials of CpG-ODNs as immunomodulators in poultry need to be 

further explored. 

Analysis of swine skin-derived dendritic cells in the induction of innate 

immune responses to Foot-and-mouth disease virus (FMDV)Elida M. Bautista, 

Geoffrey S. Ferman, Douglas Gregg, Mario C. Brum, Marvin Grubman and William T. 

Golde. 

USDA, ARS, Plum Island Animal Disease Center, Greenport, NY 11944. 

To understand the role of dendritic cells (DC) in the initiation of immune responses 

against FMDV and to determine whether FMDV infection may alter this function, we 

analyzed the interaction of FMDV with skin-derived DC of swine. Innate responses of 

DC to in vitro FMDV exposure with infectious or attenuated leaderless (LL) viruses was 

determined by measuring cytokine mRNA and protein expression using Real-time PCR 

analysis and ELISA, respectively. Activation state of these DC was determined by 

2003 ARS Immunology Research Worskshop 20 
measuring surface expression of SLAII (MHC Class II) and CD80/86. SLAII was 

minimally affected and CD80/CD86 slightly reduced in DC exposed to either virus. DC 

consistently responded to both viruses with a rapid increase in IFN-beta and TNF-alpha

mRNA expression that peaked at 4-8 h post-exposure. IFN mRNA was only moderately 

increased over the constitutive levels. Analysis of supernatants (SN) of virus exposed DC 

show secretion of IFN and IFN by protein assay (ELISA), but neither protein was

detected in the control culture SN with no virus. Biological function of IFN was 

confirmed only in the virus-exposed cell culture SN by antiviral assay. Consistent with 

the antiviral effect of IFNs, no productive infection by FMDV was detected in DC. 

Immunocytochemistry and confocal microscopy of skin sections and freshly isolated DC 

demonstrated these cells constitutively express IFN These results show that DC can 

mount an innate type I IFN response to FMDV, which renders them resistant to 

productive viral replication. This study also revealed an important property of porcine 

skin DC in that these cells constitutively express and accumulate IFN protein in the 

cytoplasm, and respond to virus exposure by secreting IFN and by activating the IFN 

gene and secreting this cytokine as well. Together, our data show that skin DC have an 

important role in the innate immune response of swine to FMDV. 

2003 ARS Immunology Research Worskshop 21 
Immune Regulation 

Regulatory role of gamma/delta T cells in allergic airway hyperreactivity 

Willi Born 

National Jewish Center, Denver, CO 80206. 

Airway hyperresponsiveness to cholinergic agonists (AHR) is a 

life-threatening condition associated with many diseases of the vertebrate 

lung. Gamma/delta T cells exert a strong regulatory effect on AHR. We have 

used a mouse model of allergic airway disease to investigate the underlying 

mechanisms. Features include regulation during antigen priming and 

challenge, systemically and within the lung, and small populations of 

gamma/delta T cells with opposed regulatory influences on AHR. As 

regulatory functions co-segregate with T cell receptor Vgamma expression, 

specific ligand recognition may play a role, and ligands may be expressed on 

dendritic cells (DC). Gamma/delta T cells appear to monitor DC in the lung, 

and early signals from DC might induce their regulatory effect on the 

developing antigen-specific immune response and airway smooth muscle 

contraction. 

The contribution of memory T cells subsets to antiviral immunity in the lung 

David L. Woodland 

Trudeau Institute, Saranac Lake, NY 12983, USA 

Respiratory virus infections elicit memory CD4+

 and CD8+ T cell populations that persist 

for the life of the individual and are able to mediate accelerated clearance of secondary 

virus infections. Given the protective capacity of these cells, there has been substantial 

interest in developing vaccines that promote cellular immunity in the lung. However, we 

have little understanding of how T cell memory is established, maintained, and recalled 

in peripheral organs. To begin to address these issues, we have been analyzing memory

T cell populations elicited by influenza and parainfluenza virus infections in a mouse 

model. We show that memory T cells are extremely heterogeneous in terms of their 

anatomical distribution, phenotype, function, and longevity. Moreover, distinct 

subpopulations vary greatly in their capacity to respond to infection, depending on their 

initial phenotype and anatomical location. These observations have significant 

implications for the development of vaccines designed to promote cellular immunity in 

the lung. 

2003 ARS Immunology Research Worskshop 22 
Regulation of the CD4+

 T lymphocyte response by Anaplasma marginale.

Wendy C. Brown, Guy H. Palmer, and Jeffrey R. Abbott 

Department of Veterinary Microbiology and Pathology, Washington State University, 

Pullman, WA 99164-7040. 

MSP2 is serologically immunodominant for individuals infected with Anaplasma. In 

cattle protected by immunization with A. marginale outer membranes, CD4+

 T cell and 

IgG responses are also predominantly directed against MSP2 and MSP3, which share the 

conserved carboxy region. In fact, MSP2-specific T cells also recognize MSP3. Antigenic 

variation in MSP2 occurs throughout the lifespan of persistently infected but clinically 

healthy cattle, and sequentially emerging variants are controlled by the immune response. 

Both variant-specific CD4+

 T cells and antibody are believed to be important for this 

control. We hypothesized that immunization with multiple MSP2 variants should provide 

protection against challenge with organisms expressing the same MSP2 variants. To test 

this, eight calves were immunized with native MSP2 using either IL-12 or CpG DNA 

adjuvants that stimulated IgG and type 1 T cell responses, and together with four control 

animals, were challenged five months later with homologous organisms that expressed 

the same major MSP2 variants. In spite of developing significant and strong MSP2-

specific CD4+

 T cell proliferative, IFN-γ, and IgG responses, vaccinates were not 

protected from challenge. Importantly, acute infection was not due to the emergence of 

novel MSP2 variants. To elucidate the reasons for the lack of protection in the vaccinates, 

we compared antigen-specific responses of CD4+

 T-lymphocytes from MSP2 vaccinates 

and control cattle cryopreserved before and after A. marginale challenge using IFN-γ

ELISPOT and proliferation assays. Reproducibly strong MSP2-specific PBMC responses 

were detected from immunized animals prior to challenge and for two weeks after 

challenge, which was prior to peak rickettsemia. Surprisingly, MSP2-specific responses 

of PBMC collected during the peak of rickettsemia were undetectable in all animals. This 

severely depressed MSP2-specific T cell response was still observed two-four months 

later, and was also apparent when using A. marginale homogenate. In contrast, the 

response to unrelated Clostridium vaccine antigens was not ablated during acute 

infection. By four to seven months after peak rickettsemia, MSP2-specific T cell 

responses were detected in several vaccinates; however, these remained significantly 

depressed. Our data suggest that in addition to immune evasion via antigenic variation in 

surface proteins, A. marginale induced unresponsiveness may facilitate organism

persistence. Potential mechanisms include induction of regulatory T cells, induction of T 

cell anergy, and exhaustion of MSP2-specific T cells. Supported by USDA NRI grants 

99-35204-8368 and 02-35204-12352 and NIH NIAID awards K08-AI49276 and R01-

AI44005. 

2003 ARS Immunology Research Worskshop 23 
Bovine T cell response to antigens of Neospora caninum.

Wenbin Tuo1

, Ray Fetterer2, Pat Boyd1, Lou Gasbarre3, JP Dubey2. 1

 USDA, ARS, Immunology and Disease Resistance Lab, 2Animal Parasitic Diseases Lab, 

3

Bovine Functional Genomics Lab, ANRI, BARC-East, Beltsville, MD 

Neospora caninum is an apicomplexan parasite that infects a number of cell types, 

causing neosporosis in several animal species including cattle. In the past decade, 

neosporosis has been recognized as a primary cause of abortion in cattle worldwide. It 

was estimated that approximately 20% of all bovine abortions are attributed to 

neosporosis; Neospora-related abortion costs the cattle industry in excess of $35 million a 

year in the state of California alone. In infected cows, N. caninum elicits a detectable 

antibody (Ab) response as shown by extensive studies, whereas N. caninum-induced cellmediated immunity (CMI) is poorly understood. Previous exposure to, or infection by, 

the parasite does not provide protection against abortion or vertical transmission, in spite 

of the presence of Neospora-specific Ab in maternal circulation. The objective of this 

study was to define Neospora-specific CMI in cows infected with N. caninum and to 

identify immunodominant T helper cell Ag that may be used as vaccine candidates. Total 

Neospora Ag was prepared by freezing and thawing or sonication of N. caninum

tachyzoites grown in a monkey kidney or a bovine macrophage cell line. Peripheral 

blood lymphocytes (PBL) from Neospora infected cows were stimulated with total 

soluble Neospora Ag weekly in the presence of irradiated PBL as a source of Ag 

presenting cells. Flow cytometry analysis showed 90-98% of these Neospora Ag specific 

T cells (week 3-8) were CD2+

CD3+CD4+. A T cell proliferation assay was established 

and used to determine Ag specificity of Ag-specific T helper cells and antigenic activity 

of Neospora Ag. A bovine specific IFN-( ELISA was used to determine IFN-( content 

in supernatants of T cells stimulated with Neospora Ag. The total Neospora tachyzoite 

Ag was separated into 5 fractions by anion exchange using HPLC. Results indicate that 

high levels of IFN-( were produced by both PBL and T cell lines specific to Neospora 

Ag. Using the T cell proliferation assay, we showed that proteins from HPLC Fractions 1 

and 2 had low or undetectable T cell-stimulating activity, whereas proteins from

Fractions 4-6 had higher stimulatory activity. However, Fraction 5 contained the highest 

antigenic activity in all T cell lines assayed. Western blot analysis showed that Fraction 5 

contained 8-10 distinctive Neospora protein bands that were defined by a rabbit 

antiserum raised against to Neospora tachyzoites (NC-1 strain). These studies suggest 

that Ag-specific T helper cells (CD4+

 CD3+) may be used to screen for immunodominant 

Neospora Ag. Neospora proteins that specifically stimulate T helper cell proliferation 

and cytokine production identified using our T cell proliferation and cytokine production 

assays may be used as vaccine candidates. This research will enhance our understanding 

of bovine CMI to Neospora infection and may facilitate the development of a vaccine

against bovine neosporosis. 

2003 ARS Immunology Research Worskshop 24 
Progress towards developing effective subunit vaccine responses by CD8+ 

memory and effector T cells in a bovine model of tuberculosis.

D. Mark Estes 

University of Texas Medical Branch, Department of Pediatrics and Pathology, The Sealy 

Center for Vaccine Development 

Cell-mediated immunity (CMI) is a key component of the protective immune response 

for many intracellular pathogens (viruses, bacteria and parasites). CD8+ cytotoxic T cells 

(hereafter referred to as CTL) are a primary effector cell type in the CMI response to 

intracellular pathogens via their ability to lyse infected target cells and produce key 

regulatory and effector cytokines. CD8+ T cells recognize relatively short peptides (8-11 

amino acids in length) presented in the context of MHC class I molecules on the surface

of an antigen presenting cell (APC). APC processing of polypeptides from both inside 

and outside of the cell is the first step in the formation of MHC class I-peptide complexes 

and the generation of T cell responses. Rational design of vaccines for intracellular 

pathogens requires knowledge of both this process and the means by which costimulatory 

signals are delivered to naïve T cells by APC. Costimulatory signals are required in 

addition to T cell receptor (TCR) engagement to drive the response. The requirements for 

accessory molecule costimulation (collectively, signal 2) in the activation of naïve CD8+ 

T cells, in addition to the primary signal (signal 1) generated via the T cell receptorantigen-MHC complex, are not well understood, particularly in cattle. Knowledge of this 

important issue is essential to the development of viable vaccine strategies that involve 

elicitation of effective CD8+ T cell responses. 

Effects of the mammary gland on the composition and function of peripheral 

blood mononuclear leukocyte populations in periparturient dairy cows.

J. P. Goff, K. Kimura, B. J. Nonnecke 

USDA, ARS, Periparturient Disease of Cattle Research Unit, National Animal Disease 

Center, Ames, IA 50010 

During the periparturient period, the dairy cow experiences a state of natural 

immunosuppression that is associated with heightened susceptibility to infectious disease. 

During this period, alterations in T cell percentages in the circulation parallel a reduction 

in functional capacities of peripheral blood mononuclear cell (PBMC) population. 

Hormonal and metabolic demands of pregnancy and lactation as well as the physiologic 

stress of calving likely contribute to periparturient immunosuppression. Objectives of this 

study were to evaluate the phenotype and functional capacities of PBMC populations in 

periparturient cows, and secondly, determine whether changes in composition and 

function could be, in part, attributable to the presence of a functional mammary gland. 

Ten mastectomized and eight intact multiparous Jersey cows (six for each for functional 

study) were used. Using monoclonal antibodies against T-cell subsets, the B-cell, and the 

monocyte, the composition of PBMC populations was characterized by flow cytometry. 

For the analyses, cells were obtained several times a week from –4 wk prepartum to 4 wk 

postpartum. Functionality of PBMC populations was assessed by assays measuring 

mitogen-induced lymphocyte proliferation, IgM secretion, and IFN-( secretion. Cells 

used in these assays were collected twice weekly from –3 wk prepartum to 3 wk 

2003 ARS Immunology Research Worskshop 25 
postpartum. Percentages of CD3+, CD4+, CD8+, and γδ TCR+ cells in intact cows 

decreased progressively with the approach of parturition, while monocyte percentages 

increased. Synthesis of DNA and secretion of IFN-( and IgM by mitogen-stimulated 

intact-cow PBMC were lowest around the time of calving. In contrast, compositional and 

functional changes in PBMC populations from mastectomized cows were minimal or 

absent at these times. Regression analysis revealed significant associations between 

composition and functional capacity of the PBMC population. These results indicate that 

the mammary gland and metabolic stresses associated with lactation contribute to the 

generalized immunosuppression affecting dairy cows during the periparturient period. 

2003 ARS Immunology Research Worskshop 26 
Immunogenetics 

Accelerating vaccine design: new in silico epitope mapping methods

Annie De Groot, M.D. 

CEO, EpiVax, Associate Professor, Brown Medical School 

T cell responses play an important role in immunity to parasites and other microbial 

agents of infectious diseases, therefore a number of T cell-directed vaccines are in 

development. Computer-driven algorithms that facilitate the discovery of T cell epitopes 

from protein and genome sequences are now being used to accelerate the development of 

human vaccines. Similar tools are now becoming available for for animal vaccines. My

group recently developed new in silico methods for predicting epitopes for cattle and 

swine; the development process and validation studies will be described in this talk. 

Epitope-mapping algorithms offer a significant advantage over other methods of epitope 

selection, such as the screening of synthetic overlapping peptides, because high 

throughput screening can be performed in silico, followed by ex vivo confirmatory 

studies. Using epitope-mapping algorithms, putative T cell epitopes can be derived 

directly from genomic sequences, allowing researchers to circumvent labor-intensive 

cloning steps in the genome-to-vaccine discovery pathway. 

Marek's disease and the MHC: How viruses subvert the immune system for 

their advantage.

H. Hunt, H. Cheng, and H. Zhang 

USDA, ARS, Avian Diseases and Oncology Laboratory, East Lansing, MI 

Immune response genes of the chicken play a critical role in the resistance or 

susceptibility to Marek’s disease (MD), a T-cell tumor induced by a herpesvirus named 

MD virus (MDV). The major histocompatibility complex (MHC) is one component of 

the immune response that has a major influence on the outcome of MD as well as the 

immune protection afforded by MDV vaccines. The USDA-ARS Avian Disease and 

Oncology Laboratory (ADOL) in East Lansing, MI is investigating the influence of the 

chicken MHC and other genetic systems involved in the immune response to MD. 

ADOL’s approach is multidisciplinary incorporating traditional genetics, functional 

genomics/proteomic and immunologic analysis to dissect the host-pathogen interactions 

responsible for resistance or susceptibility to MDV. 

The traditional genetics approach involves the generation of recombinant congenic 

chicken strains (RCS). ADOL White Leghorn lines 6 and 7 are resistant or susceptible to 

MDV, respectively, and differ in immune response characteristics and lymphoid organ 

size. The two lines were intermated and full-sib mated to generate 19 inbred sub-lines 

designated 6C.7A through X. Each 6C.7 sub–line consists of approximately 12% of the 

susceptible line 7 genome inserted into the resistant line 6 genomic background. The 

immune response, disease resistance and genetic phenotypes of these 6C.7 RCS are 

2003 ARS Immunology Research Worskshop 27 
currently under investigation at ADOL using gene mapping and immune assay 

approaches. 

MDV, like other herpesviruses, has the ability to alter the expression of antigen 

presenting glycoproteins encoded by the MHC. MDV inhibits the cell surface expression 

of MHC class I glycoproteins but, unique to MDV, induces the expression of MHC class 

II glycoproteins. ADOL’s multidisciplinary approach incorporating traditional 

immunologic assays coupled with genomics and proteomics is beginning to unravel the 

host pathogen interactions responsible for these observations. 

Expression of specific MHC Class II DRB1 alleles associates with sheep 

persistently infected with ovine progressive pneumonia virus (OPPV).

L.M. Herrmann1, W.P. Cheevers2, D.P. Knowles

1,2

1

USDA, ARS, Animal Disease Research Unit, Pullman, WA 99164-6630; 2 Department 

of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 

99164-7030. 

During OPPV infection in a flock, there is a clear dichotomy where a few sheep produce 

clinical signs and most become persistently infected without clinical signs. In long-term

persistent infections, cell-adaptive immunity plays the significant role in controlling virus 

replication. In other small ruminant lentiviruses, CD4 positive and not CD8 positive T 

lymphocytes are required for adequate proliferation upon exposure to viral antigens 

(Perry et al., 1995). This along with two other studies indicating that the immunogenetics 

of the host dictate the course of infection and the antibody response to OPPV (de la 

Concha-Bermejillo et al., 1995; Cheevers at el., 1999), support the idea that MHC Class 

II immunogenetics play a significant role in directing disease outcome. Since ovine 

MHC Class II DRB1 encodes and expresses the antigen presentation domain on the 

antigen-presenting cell, we evaluated the allelic expression of DRB1 in peripheral blood 

mononuclear cells (PBMC) from 10 sheep persistently infected with OPPV. Five of 

these sheep have high OPPV loads, and five have low OPPV loads by real time PCR of 

integrated DNA in PBMC. The relationship between viral load and specific DRB1 allele 

expression will be discussed. In addition, the relationship of viral genetics in terms of the 

surface envelope glycoprotein (SU) and specific DRB1 allele expression will also be 

discussed. 

From basic porcine immunology to practical application

John E. Butler 

University of Iowa 

Characterizing the elements that comprises the immune system is the first step needed to 

provide the information and tools for ultimately determining the role of the immune 

system in health and disease. Described here is a 12-year journey of discovery in a farm 

animal with a pattern of passive immunity and a precosial nature that makes it an ideal 

model for providing insight into immunological development, receptor repertoires and 

the immunopathology of an important viral disease. 

2003 ARS Immunology Research Worskshop 28 
Basic investigations revealed the swine immune system is anything but a carbon copy of 

the human and mouse systems. For example, swine rely on only 10 combinatorial 

rearrangements, not 9,000, to develop their heavy chain antibody repertoire. Equally 

efficient is the development of the V-kappa repertoire that primarily uses one V kappa 

gene family and one J kappa gene segment. Also in contrast to mice and humans, early B 

cell development is adult-like, and surprisingly involves the thymus beginning at 

midgestation, where switch recombination is pronounced in utero in the absence of 

environmental antigen. Naïve B cells migrate to the ileal Peyers patches, each follicle 

containing the diversified descendents of a single B cell clone. 

While B cell development occurs rapidly in late gestation, the ability of the newborn to 

mount an immune response requires exposure to colonizing bacteria or infective virus 

that produce the ligands for the Toll-like receptors (TLRs). 

While the mechanism of antibody repertoire development in swine distinguish it from

rodents and primates, T-cell development follows a conserved evolutionary pattern that 

parallels that in other mammals. The TCR beta repertoire strongly resembles that of 

humans and mice in both genome organization and the presence many homologous Vbeta families. The V-epsilon repertoire is also conserved and an invariant V epsilon 

transcript occurs early in fetal life reminiscent of that in mice. 

We were able to use the tools of discovery and knowledge we had gained to employ the 

isolator piglet model to show that PRRSV directly causes a B cell lymphoproliferative 

disorder characterized by immune dysfunction, autoimmunity, immune complex 

deposition and kidney and vascular damage. We hope as the journey continues, we shall 

soon show how PRRSV alters the T cells repertoire and how specific bacterial- and viralderived TLR ligands allow the adaptive immune system to develop. 

2003 ARS Immunology Research Worskshop 29 
December 3, 2003

Immunity and Nutrition 

The effect of human-derived probiotic bacteria on the immune and intestinal 

function of pigs

Gloria Solano-Aguilar*1, Harry Dawson1, Tonya Ledbetter1, Terez Shea-Donohue1, 

Norberta Schoene1

, Jeff Call

2

, Ethiopia Beshah3, William Hare3 and Joseph Urban1. 

USDA, ARS, 1

Beltsville Human Nutrition Research Center, Beltsville MD; 

2 Eastern 

Regional Research Center, Wyndmoor, PA 19038; 3

Animal &Natural Resources 

Institute; Beltsville, MD 20705. 

Probiotics are specific microorganisms that can establish and grow in a compartment of 

an exposed host and provide some positive health benefit; strains of Lactobacilli, 

Bifidobacteria and Enterococcus are commonly used probiotics. The benefits claimed 

vary from protection against pathogenic microorganisms and viruses, stimulation of the 

intestinal immune function and enhanced disease resistance. However, the basis of these 

claims is often confounded by a lack of demonstrable growth and function of the 

probiotic strain in the gut. Our objective was to test if dietary probiotics can establish in 

the pig and affect immunity to an infectious agent. 

Experiment 1: 

A pilot study using eight 12-week-old pigs was performed to determine if animals fed 

with a daily dose of 10 billion colony forming units (cfu) of freeze-dried Lactobacillus 

rhamnosus GG (LGG; Culturelle, Conagra, NE) would show some change in their 

immune response. Whole blood for isolation of peripheral blood mononuclear cells 

(PBMC) and fecal samples for DNA extraction were collected throughout the experiment 

at days 4, 7, 12, 18 and 25 after initiation of dietary treatment. At the end of the 

experiment (day 26) pigs were euthanized and tissues from different mucosal sites were 

collected for: 1) cell immune phenotype analysis by flow cytometry; 2) DNA isolation 

and identification of LGG strain in intestinal mucosa and fecal samples, 3) measurement 

of IFN-( production in vitro after mitogen stimulation, and 4) mRNA expression of a 

panel of immune markers by real-time PCR. PBMC isolated from LGG-exposed pigs 

produced IFN-( in response to Con A stimulation, while IFN-( was barely detectable 

(assay range in ng /ml) in culture supernatant of PBMC isolated from unexposed pigs. 

LGG-specific fluorogenic probes were able to detect LGG DNA in fecal samples of 

treated animals as early as 4 days post-treatment. Mucosal tissue samples and fecal 

contents taken from intestinal sites of LGG treated animals indicated that LGG DNA was 

detected at 4-fold higher levels in the proximal colon compared to distal colon and 

cecum. No signal was detected in jejunum or ileum of any animals or in samples from

non-treated animals. Real time PCR analysis indicated a selective low level increase in 

gene expression for Th1-derived type 1 immune cytokines in localized mucosal sites after 

probiotic delivery. 

2003 ARS Immunology Research Worskshop 30 
Experiment 2: 

To test the effect of early colonization of probiotic bacteria on the immune response to a 

nematode parasite infection, Bifidobacterium lactis (Bb12)(Mak Wood Inc, WI) (10 x 

1010 cfu) was administered to 2 sows starting at the last third of pregnancy through 

weaning of their piglets at day 28; two other sows and their litters were left as untreated 

controls. After birth, Bb12 capsules were also administered to pigs born to treated sows. 

Pigs born from untreated sows were maintained as untreated controls. All piglets were 

inoculated with Ascaris suum eggs at 6 weeks of age and tissue samples were collected 

for functional gut in Ussing chambers 3 weeks later. Probiotic treatment did not alter 

normal absorption of glucose in the small intestine, but did attenuate A. suum-induced 

inhibition of glucose absorption that correlates with expulsion of the worm from the pig 

intestine. 

Conclusions: 

Human derived probiotics can modulate immune function and selectively affect local 

responses to parasitic infection while promoting swine health. This model can be 

extended to assess the activity of selected probiotics on pig responses to other infectious 

agents and allergens that negatively affect pig production. 

The care and feeding of an immune system: How much is needed and what 

are the priorities?

Kirk C. Klasing 

Department of Animal Science, University of California, Davis CA 95616. 

How big is the immune system and how good is it at competing for scarce nutrients? 

Immunologists have neglected to measure the size of the immune system. Nutritionists 

have neglected to determine the nutritional requirements of the processes that we call 

‘immunity’. Currently, these first principles of nutrition and immunity can only be 

estimated from a synthesis of experiments that were designed for other reasons. We need 

to know: the costs of developing an immune system that is sufficiently robust to be 

dependably protective; the costs of maintaining this system of leukocytes in large enough 

copy number to be effective; and the cost of using this system when challenged the host 

is challenged by a proficient pathogen. A preliminary analysis of these costs suggests that 

development of innate immunity is cheap but development of adaptive systems 

(lymphocytes) is relatively expensive. The reverse is true for maintenance. Following a 

challenge, the production of immunoglobulin and the clonal expansion needed for 

protective T-cell responses are cheap to generate but can cause immunopathology in 

defending against some pathogens. The systemic sequelae of an acute phase response are 

especially expensive and markedly impact the nutritional state and productivity of an 

animal. The types of nutrient transporters utilized by leukocytes determine their priorities 

for nutrients. Macrophages have a high priority, B-lymphocytes are intermediate, and Tlymphocytes are inept. 

2003 ARS Immunology Research Worskshop 31 
Aging, immune response, and infectious disease: molecular mechanisms and 

reversal by nutrient intervention.

Simin Nikbin Meydani, DVM, PhD, Dayong Wu, MD, PhD, Sung Nim Han, PhD, RD, 

Raina T. Gay. 

USDA, ARS, Nutritional Immunology Laboratory, Jean Mayer Human Nutrition 

Research Center on Aging at Tufts University, Boston , MA. 

The incidence of neoplastic and infectious diseases is increased in the elderly, as is the 

resulting morbidity and mortality. The well-documented age-associated dysregulation of 

immune function is an important contributor to the increased incidence of mortality from

these diseases. Accumulating evidence indicates that T cell-mediated immune responses 

decline with aging. We, as well as others, have demonstrated that in addition to intrinsic 

changes in T cells, increased macrophage (Mφ)–derived prostaglandin E2 (PGE2)

production contributes to the age-associated decline in T cell function [1]. We have 

previously shown that vitamin E supplementation of aged mice and elderly humans 

significantly improves T cell mediated functions including their response to vaccination 

[2-4]. Vitamin E exerted its immunostimulatory effect by two distinct mechanisms: 1) by 

reducing Mφ PGE2 production, and 2) by a direct effect on T cell function. We further 

showed that vitamin E enhances T cell function directly by enhancing the cell dividing 

and IL-2 producing capacity of naïve T cells from old mice, with no effect on memory T 

cells [5]. 

To determine the clinical significance of vitamin E-induced immune enhancement in the 

aged we investigated the effect of vitamin E supplementation on viral and bacterial 

infection in the aged. Vitamin E supplementation of aged mice significantly reduced lung 

viral titer in mice infected with influenza virus.[6, 7]. On the other hand, old mice fed a 

diet supplemented with vitamin E (500 ppm) compared to those fed a diet adequate in 

vitamin E, vitamin E supplementation did not protect against primary pulmonary 

Staphylococcus aureus (S. aureus) infection, whereas it was protective against secondary 

pulmonary S. aureus infection following influenza infection. This was indicated by a 

significantly lower bacterial count in the lungs of vitamin E-supplemented mice 

compared to that in control mice infected with S. aureus following influenza infection. In 

a recently completed double-blind, and placebo controlled study, we determined the 

effect of vitamin E on respiratory infection in nursing home residents. The results of this 

study as well as others will be discussed. 

This work was supported in part by Federal funds from USDA, Agriculture Research 

Service under contract number 58-1950-9-00, and from the National Institute on Aging, 

grant AG 09140-09. 

References: 

Beharka, A.A., et al., Macrophage PGE2 production contributes to the ageassociated decrease in T cell function which is reversed by dietary antioxidants.

Mech. Ageing Dev., 1997. 93: p. 59-77. 

2003 ARS Immunology Research Worskshop 32 
Meydani, S.N., et al., Vitamin E supplementation suppresses prostaglandine E2

synthesis and enhances the immune response of aged mice. Mech. Ageing Dev., 

1986. 34: p. 191-201. 

Meydani, S.N., et al., Vitamin E supplementation enhances cell-mediated 

immunity in healthy elderly subjects. Am J Clin Nutr, 1990. 52: p. 557-563. 

Meydani, S.N., et al., Vitamin E supplementation and in vivo immune response in 

healthy elderly subjects. A randomized controlled trial. JAMA, 1997. 277: p. 

1380-1386. 

Adolfsson, O., B.T. Huber, and S.N. Meydani, Vitamin E-enhanced IL-2 

production in old mice: naive but not memory T cells show increased cell division 

cycling and IL-2-producing capacity. J Immunol, 2001. 167(7): p. 3809-17. 

Han, S.N. and S.N. Meydani, Antioxidants, cytokines, and influenza infection in 

aged mice and elderly humans. J. Infectious Disease, 2000. 182: p. 574-581. 

Hayek, M.G., et al., Vitamin E supplementation decreases lung virus titers in mice 

infected with influenza. J. Infect. Dis., 1997. 176: p. 273-276. 

Nutritional plane affects antigen-induced proliferation of lymphocyte subsets 

in milk replacer-fed calves vaccinated with M. bovis bacillus Calmette-Guerin 

(BCG). 

B. J. Nonnecke1

, M. R. Foote2, M .A. Fowler3, B.L. Miller

3

, W. R. Waters4

1

 USDA, ARS, Periparturient Diseases of Cattle Research Unit, National Animal Disease 

Center, Ames, IA, 2

Iowa State University, Ames, IA, 3Land O’ Lakes, Inc., Webster City, 

IA and 4

USDA, ARS, Bacterial Diseases of Livestock Research Unit, NADC, Ames, IA. 

National Research Council recommended nutrient requirements for the neonatal dairy 

calf do not support optimal growth, and may compromise immune function. In a 

preliminary study, dietary energy and protein were shown to modulate in vitro production 

of interferon-gamma and nitric oxide by polyclonally stimulated blood lymphocytes from

milk replacer-fed calves. We also have shown that adaptive (i.e. Th1-dependent) 

immune responses of milk replacer-fed neonatal calves vaccinated with Mycobacterium 

bovis bacillus Calmette Guerin (BCG) and challenged with purified protein derivative 

(PPD) are comparable in magnitude to adult responses. Using this sensitization/challenge 

model, effects of nutritional plane on antigen-induced proliferation of T cells from milk 

replacer-fed calves were evaluated. Holstein bull calves were fed a standard (ST, 0.57 

kg/d of a 22% crude protein, 20 % fat, n=4) milk replacer or an intensified (INT, 1.14 

kg/d of a 28% crude protein, 20% fat milk replacer, n=4) milk replacer for 7 wk. Calves 

and four juvenile steers (5-6 m of age) were vaccinated with M. bovis BCG at the 

initiation of the study. During the experimental period, average daily weight gain was 

greater for INT than ST calves (0.68 and 0.32 kg/d, respectively). Proliferation of CD4+

, 

CD8+

, and γδTCR+ T cell subsets in PPD- and mitogen [pokeweed mitogen (PWM) and 

2003 ARS Immunology Research Worskshop 33 
concanavalin A (ConA)]-stimulated cultures of peripheral blood mononuclear cells 

(PBMC) acquired before and 7 wk after vaccination were evaluated using flow 

cytometry. Recall responses to antigen were absent in all treatment groups prior to 

vaccination. Seven weeks after vaccination, PPD-induced proliferation of CD4+

 and 

γδTCR+ cells from vaccinated calves exceeded responses of T cell subsets from

vaccinated steers. In addition, antigen-induced proliferative responses CD8+

 cells from

INT calves exceeded those of ST calves. Lymphocytes from young adults were generally 

more responsive to mitogenic (i.e. ConA and PWM) stimulation than were cells from 

neonates. Overall, these results suggest that the proliferation of T cell subsets essential in 

CMI is affected by nutritional plane during the neonatal period. 

Real-time PCR arrays can delineate immunological and nutrition-related 

gene expression in swine. Harry Dawson 

USDA, ARS, Nutrient Requirements & Functions Laboratory, Beltsville Human 

Nutrition Research Center, Beltsville, MD. 

Real-time PCR is commonly used to verify micro array expression data. We have 

designed over 300 human and swine real-time PCR assays for large-scale primary 

analysis of genes involved in immunological or nutritional function. These assays have a 

greater linearity, specificity and sensitivity when compared to micro array. The 

immunological foci of these assays are on pathways involved in apoptosis, toll receptor 

signaling and Th1/Th2 differentiation. The nutritional foci are on genes involved in iron, 

selenium, vitamin A and vitamin D metabolism. In conjunction with these assays, we

have developed an annotated database for data mining using micro array analysis 

software packages. Using subsets of these assays, we profiled pig responses to 

Toxoplasma gondii, Ascaris suum, , Oesophagostomum dentatum, Salmonella 

typhimurium and Campylobacter jejuni. Toxoplasma elicited a generalized Th1 response 

typified by high-level induction of IFN-(, TNF-" and iNOS. Trichuris induced a limited 

Th2-associated response including localized increased IgG, IgG1, IgG4, IL-13, IL13Ra2, SOCS3, and CHIA expression. Resistant animals had higher GATA-3, IgE, IL10, CD3e, CD40L, CD86, and MCP-2 expression, and lower ARG1 and MUC5AC 

expression. The impact of this tool lies in the expanding access to genetic information 

between mammalian species allowing hypothesis testing of infection resistance 

mechanisms using quantitative expression analysis of large numbers of genes at multiple 

tissue sites. There is also potential application to the measurement of therapeutic efficacy 

for a variety of immunological and nutritional interventions against human and swine 

diseases. Ongoing work includes studies with vitamin A and its derivatives. 

2003 ARS Immunology Research Worskshop 34 
Immunity and Disease Resistance 

Mechanisms of innate and adaptive immunity in swine

Michael Murtaugh, Zhengguo Xiao, and Mark Rutherford. 

University of Minnesota, College of Veterinary Medicine, Veterinary Pathobiology, St. 

Paul, MN 

Mouse models of immune mechanisms of infectious disease resistance may be unsuitable 

for livestock species. The porcine immune system, for example, differs in immune tissue 

architecture, T cell phenotypes, cytokine expression patterns, cytokine activities, and 

possibly gene expression profiles. Thus, genomic, molecular, cell biologic, and other 

direct approaches will be required to elucidate porcine protective immune responses to 

infectious agents. Studies of immunity to porcine reproductive and respiratory syndrome 

virus (PRRSV), currently the most important pathogen of swine worldwide, illustrates the 

current limitations in the field. PRRSV exclusively infects macrophages of swine and 

causes a prolonged acute and persistent infection of tissues rich in macrophages. Detailed 

analysis of nonspecific innate and specific acquired immunity to PRRSV is beginning to 

reveal heretofore unsuspected mechanisms for control and elimination of viral infection 

in animals. PRRSV appears to subvert host defense by infection of lung macrophages 

without provoking the characteristic innate interferon α/β response to RNA viruses. The 

appearance of neutralizing antibodies does not occur until about three weeks after 

infection; acute infection manifested as viremia lasts for 4-5 weeks, and infectious virions 

are detected in lymphoid tissues for 3-4 months thereafter. While infection lingers for a 

prolonged time, viral load in lymphoid tissues plummets dramatically in the early phases 

of persistent infection. During this period, the PRRSV-specific T cell response at sites of 

infection is weak, transient, variable, and independent of local viral load. Surprisingly, 

pigs are largely resistant to reinfection with homologous or heterologous viral isolates 

even though neither humoral nor T-cell anamnestic responses are observed. We

hypothesize that PRRSV control in swine is due primarily to a reduction in permissive 

macrophages, and secondarily to specific humoral and cellular factors. Further 

investigation of immunological mechanisms that might provide insights to development 

of effective vaccines and immunotherapeutic strategies will require reagents that 

stimulate antigen-specific T cells and B cells and assays that are both specific and 

functionally relevant. Achieving these goals will require systems for expression and 

presentation of individual proteins or polypeptide fragments, tetramers, cytokine-specific 

ELISA, ELISPOT and FACS assays, and a better understanding of basic immunology of 

livestock species. Genomic identification of the gene repertoire involved in innate and 

adaptive immunity to specific pathogens and of the allelic variation in key regulatory and 

effector genes will be required to fully characterize basic immunological processes. Our 

initial foray into this area, identification of genes differentially expressed in porcine 

Peyer’s patch, revealed that 45% of expressed sequences have no known function or are 

novel genes. Genes of unknown function also are differentially expressed in macrophages 

infected with PRRSV. These results indicate that the biochemical and cell biologic 

mechanisms responsible for immunity in swine, to a substantial degree, remain to be 

2003 ARS Immunology Research Worskshop 35 
discovered. A better grasp of this knowledge will be required for rational design and 

genetic selection of disease resistant animals. 

Genetic and immunologic basis for resistance to GI nematode infections

Louis C. Gasbarre, Tad Sonstegard, Ricardo Araujo, and Curt Van Tassell 

USDA, ARS, Bovine Functional Genomics Laboratory, ANRI, BARC, Beltsville MD 

Gastrointestinal nematode infections of cattle are a constraint on the efficient raising of

cattle throughout the world. Most of the nematodes species found in cattle stimulate a 

level of protective immunity that reduces the number of worms that develop after 

secondary challenge, but cattle remain susceptible to reinfection by Ostertagia for a 

prolonged period of time (i.e. 2 years or more). This prolonged susceptibility to 

reinfection is a major reason that this parasite species remains the most economically 

important GI nematode in temperate regions throughout the world. Although, individual 

animals remain susceptible to reinfection for a prolonged period of time, there are a 

number of manifestations of the immune response that result in an enhanced level of herd 

immunity, and the overall result of these manifestations of immunity is a reduction in 

parasite transmission within the cattle herd. In cattle, the immune response seen in the 

abomasum shows high levels of expression of IL4 in the draining lymph nodes and in 

lymphocytes isolated from the mucosa, but effector cell populations in the tissues 

surrounding the parasites, are not typical, inferring that Ostertagia has evolved means to 

suppress or evade protective immune mechanisms, including the synthesis and release of 

immunosuppressive substances. The immune responses elicited by GI nematode 

infections are moderately influenced by host genetics, and a small percentage of 

individual animals in a given herd are responsible for the majority of parasite 

transmission. In general, cattle can be separated into three types: 1) those which never 

demonstrate high parasite numbers, 2) those that show increasing signs of infection 

during the first 2-3 months on pasture and then develop responses that regulate parasite 

numbers and or parasite fecundity, and 3) those which continue to maintain high parasite 

burdens. Studies are in progress to identify the genes controlling resistance. Two 

approaches are being utilized. The first involves mapping of quantitative trait loci (QTL) 

that influence the level of parasites infection. For these studies a population of cattle has 

been developed in which careful phenotypic characterization of calves has been 

combined with a complete genome scan to identify polymorphic markers that segregate 

with either increased or diminished resistance to infection. Using this approach QTL 

that affect parasite resistance have been demonstrated on 8 different bovine 

chromosomes. The second approach utilizes microarray technology to assess differential 

gene expression in resistant versus susceptible cattle. We have developed a 400+ 

candidate gene array and have used it to look at gene expression in the local tissues of 

infected cattle. Results to date have indicated marked differences in gene expression 

patterns in genes associated with both adaptive and innate immune responses. Current 

work is focused on validation of these results by real time PCR, the development of 

global arrays for a more thorough look at the gene expression patterns, and integration of 

the QTL and array technologies for identification of important genes. 

2003 ARS Immunology Research Worskshop 36 
Immunity and Disease Resistance

Heterophils isolated from chickens resistant to extraintestinal Salmonella 

enteritidis infection express higher levels of pro-inflammatory cytokine 

mRNA following infection than heterophils from susceptible chickens

Pamela J. Ferro1

, Christina L. Swaggerty2, Pete Kaiser3, Igal Y. Pevzner4 and 

Michael H. Kogut2

1

Texas A&M University, Poultry Science, College Station, TX 77843, USA 

2

USDA, ARS, SPARC, 2881 F&B Road, College Station, TX 77845, USA 

3

Institute for Animal Health, Compton, Berkshire RG20 7NN, UK 

4

Cobb-Vantress Inc., Siloam Springs, AR 72761-1030, USA 

Previous studies in our laboratory have shown differences in in vitro heterophil function 

between two pure lines of broiler chicks and their F1 reciprocal crosses. The objectives 

of the current study were to 1) determine if the previously reported in vitro differences in 

heterophil function between the lines translated to an increase in in vivo resistance to

extraintestinal infection with Salmonella enteritidis (SE) and 2) to quantitate proinflammatory cytokine mRNA expression levels in heterophils isolated from SE-infected 

chicks from different lines. Day-old chicks from four different lines (pure lines=A and B; 

line C = male B × female A; line D = male A × female B) were challenged orally with 

SE. Twenty-four hours post-challenge, livers and spleens were asceptically removed 

from each chick and incubated overnight in tetrathionate enrichment broth. Enrichment 

cultures were plated and subsequently analyzed for the presence of non-lactose 

fermenting Salmonella colonies. The two lines reported to have more functionally 

efficient heterophils (A and D) were less susceptible to extraintestinal SE infections 

whereas the two lines reported to have functionally less efficient heterophils (B and C) 

were more susceptible to SE organ invasion. Four hours post-challenge peripheral blood 

heterophils were isolated from SE-infected and control chicks, total RNA extracted, and 

then analyzed for pro-inflammatory cytokine (IL-1$, IL-6, IL-8) expression using realtime quantitative RT-PCR. Pro-inflammatory cytokine mRNA expression was 

significantly up-regulated in heterophils isolated from chicks that were more resistant to 

SE organ invasion (A and D) when compared to expression in heterophils isolated from

the more susceptible lines (B and C). To our knowledge, this is the first report to 

quantitate IL-1β, IL-6, and IL-8 mRNA expression levels in heterophils isolated from

day-old chicks orally challenged with SE. These results show a relationship between in

vitro heterophil function and resistance to organ invasion by SE. These data also indicate 

that pro-inflammatory cytokine mRNA expression by heterophils contributes, at least in 

part, to resistance/susceptibility of day-old chicks to extraintestinal infection by SE. 

2003 ARS Immunology Research Worskshop 37 
High-throughput molecular approaches to identify coccidiosis disease 

resistance genes and protective immunological mechanisms in avian 

coccidiosis. 

Lillehoj

1

, Hyun S., Min1, Wongi, Matthews2, Benjamin F., and Emara3, Marlene. 

1

USDA, ARS, Animal Parasitic Diseases Laboratory, ANRI, and 2Soybean Genomics 

Improvement Laboratory, Beltsville, MD 20705, 3

Department of Animal and Food 

Sciences, University of Delaware, Newark, DE 19717 

Avian coccidiosis is caused by several Eimeria strains which infect different regions of 

the intestine inducing a strain-specific immunity. Coccidiosis usually stimulate a number 

of immunological defense mechanisms, namely both antibody- and cell-mediated. Recent 

technological advances in high-throughput molecular approaches to identify disease 

resistance genes and molecular/cell biological pathways associated with complex 

biological phenomenon now enable an alternative strategy to combat coccidiosis. We

have hypothesized that chicken genes influencing resistance to coccidiosis can be 

identified through DNA marker technology. Supporting this hypothesis, we present data 

identifying a quantitative trait locus (QTL) on chromosome 1 that significantly affects 

Eimeria oocyst shedding and three QTL that influence body weight of chickens during 

coccidiosis. One hundred and nineteen microsatellite markers, covering 80 % of the 

chicken genome with an average marker interval of 25 cM, were used for genotyping of 

F1 parents and F2 offspring from commercial broilers. The genetic mechanism of this 

locus appeared additive. The genomic scan also identified three potential growth QTL on 

Chromosomes 1, 6, and 8. These results provide the foundation for further investigation 

to validate the QTL. In addition, our recent studies on local host immunity to Eimeria

clearly indicate that intricate and complex interactions of host local cell-mediated 

immunity and parasites determine the outcome of host response to coccidiosis. Highthroughput gene expression profiling and real-time PCR have been applied to analyze 

underlying immune mechanisms controlling disease resistance/susceptibility to 

coccidiosis. The role of various cytokines whose expression increase and decrease in 

response to coccidia invasion have been studied. Recently, we have developed the first 

chicken intestinal ETS database which will be used to investigate innate and acquired 

immunities to enteric pathogens such as coccidia and Salmonella. 

2003 ARS Immunology Research Worskshop 38 
Immunointervention 

Protective killed Leptospira vaccine induces potent type 1 (Th1) immunity 

comprising responses by CD4 and WC1+ γδ t lymphocytes

C.L. Baldwin 

University of Massachusetts, Department of Veterinary and Animal Sciences, Amherst, 

MA 01003 

http://www.umass.edu/vasci/faculty/baldwin.html

 

Leptospira borgpetersenii serovar hardjo is a common cause of bovine leptospirosis and 

also causes zoonotic infections of humans. A protective killed vaccine against serovar 

hardjo was shown to induce strong antigen-specific proliferative responses by peripheral 

blood mononuclear cells (PBMC) from vaccinated cattle but not from non-vaccinated

control cattle in three studies conducted. By 7 days, up to one-third of the cells in 

cultures of sonicated leptospira and PBMC from vaccinated animals produced interferonγ (IFN-γ) as ascertained by flow cytometric analysis. The IFN-γ-producing cells included 

both WC1+ γδ T cells and CD4+

 T cells. Analysis of carboxyfluorescein succinimidyl 

ester loaded cells also showed that a similar proportion of these 2 subpopulations were 

found among the dividing cells in antigen-stimulated cultures. Infected non-vaccinated 

cattle also had an IFN-γ-based response but less than that by vaccinated animals and 

while naïve/challenged and vaccinated/challenged animals had similar levels of antigenspecific IgG1 following challenge, vaccinated animals had 1.4 fold more IgG2. In 

conclusion, while infection may induce a type 1 response we suggest it is too weak to 

prevent establishment of chronic infection. Overall the significance of these studies is the 

very potent type 1 (Th1) immune response induced and sustained following 

administration of a killed bacterial vaccine adjuvanted with aluminum hydroxide and the 

involvement of WC1 γδ T cells in the response. The response by γδ T cells is suggestive 

of a memory response since it has never occurred with cells from a large number of naïve 

cattle tested. Moreover, recent results suggest it is a specific subpopulation of the WC1+ 

cells and we hypothesize that the response by these cells may result in establishment of a 

milieu that supports development of a type 1 response by CD4 T cells. Finally, induction 

of this cellular immune response is associated with the protection afforded by the bovine 

leptospiral vaccine against L. borgpetersenii serovar hardjo although this is an 

extracellular pathogen. Similar results have been found with a second monovalent 

vaccine made with L. interrogans serovar hardjo and adjuvanted with alum while we 

have found that a traditional pentavalent vaccine containing 5 serovars of Leptospira does

not induce this response. 

Development of an oral vaccine for Escherichia coli O157:H7

Evelyn A. Dean-Nystrom 

USDA, ARS, National Animal Disease Center, Ames, IA 

Enterohemorrhagic Escherichia coli (EHEC) O157:H7, a subset of Shiga toxin-producing 

E. coli (STEC), are the most common infectious cause of bloody diarrhea in the United 

2003 ARS Immunology Research Worskshop 39 
States. A sequela of this infection, the hemolytic uremic syndrome (HUS), is the primary 

cause of acute kidney failure in U.S. children. Cattle are major sources of EHEC 

O157:H7 and other STEC pathogens associated with human infections. Our long-term 

goal is to identify ways to reduce EHEC infections in cattle, and thus reduce the risk of 

EHEC disease in humans. One approach, which is the objective of an ongoing 

collaboration with Alison O’Brien (USUHS, Bethesda, MD), is to develop an 

inexpensive, effective vaccine that will prevent cattle from becoming infected with 

EHEC. Intimin is an outer membrane protein of EHEC O157:H7 that is required for 

colonization of neonatal calves. We hypothesized that an intimin-based vaccination 

strategy in calves might reduce colonization of cattle with EHEC O157:H7. The 

demonstration that antibodies against intimin interfere with colonization and intestinal

damage in neonatal pigs supports this hypothesis. We transferred the eae gene that 

encodes for intimin into the tobacco NT-1 cell line and demonstrated that the transformed 

plant cells produce intimin, that transgenic intimin is immunogenic in mice, and that

feeding mice transgenic intimin reduced the length of time these animals shed EHEC 

O157:H7 following experimental challenge. Our collaborator, Wayne Curtis, recently 

scaled up the production of intimin-expressing NT-1 cells and the immunogenicity of 

transgenic intimin in cattle is being tested. The bacterial DNA that encodes for intimin is 

being modified so that it can be transferred into an edible plant that can be fed to calves 

and used to test the efficacy of transgenic intimin as an oral vaccine for reducing EHEC 

infections in cattle. Such a vaccine that reduces O157:H7 levels in cattle would help 

reduce EHEC infections in humans. 

Adjuvant effect of IL-1β, IL-2, IL-8, IL-15, IFN-α, IFN-γ, TGF-β4 and 

lymphotactin on DNA vaccination against Eimeria acervulina.

Wongi Min1

, Hyun S. Lillehoj

1

, and Peter Staeheli2. 1

 USDA, ARS, Animal Parasitic Diseases Laboratory, Animal and Natural Resources 

Institute, Building 1040, BARC-East, Beltsville, MD 20705 USA. 2

Department of 

Virology, University of Freiburg, D-79008, Freiburg, Germany. 

Avian coccidiosis, an economically important disease for the poultry industry, is caused 

by intracellular protozoa belonging to several species of the genus Eimeria. Coccidiosis 

results in extensive destruction of the gut epithelium accompanied by severe depression

in body weight gain, reduced feed efficiency and intestinal shedding of parasite oocysts. 

Although coccidiosis is mainly controlled by the use of chemotherapeutic agents, 

alternative control strategies are needed due to the increasing emergence of drug-resistant 

parasite strains and impending withdrawl of drug use in animal farming. Recently, novel 

vaccination strategies using antigen-encoding DNA plasmids have been shown to 

successfully induce protective cellular and humoral immune responses against a variety 

of infectious diseases. Since the emergence of gene-based immunization methodologies, 

various strategies have been used to increase the efficiency of DNA vaccination. For 

example, immune responses induced by DNA vaccination can be enhanced by cytokines. 

This study reports the first evidence of adjuvant effect of various chicken cytokines when 

given with coccidia DNA vaccine. Eight chicken cytokine genes (IL-1$, IL-2, IL-8, IL15, IFN-α, IFN-γ, TGF-β4, lymphotactin) were evaluated for their adjuvant effect on a 

suboptimal dose of an Eimeria DNA vaccine carrying the 3-1E coccidia gene (pcDNA3-

2003 ARS Immunology Research Worskshop 40 
1E). One or 10 μg of IFN-α or 10 μg of lymphotactin expressing plasmids, when given 

simultaneously with the pcDNA3-1E vaccine, significantly protected against body weight 

loss induced by E. acervulina. Parasite replication was significantly reduced in chickens 

given the pcDNA3-1E vaccine along with 1.0 μg of the IL-15, IFN-γ, TGF-β4 or 

lymphotactin plasmids or 10 μg of the IL-1β, IL-8, IL-15 or TGF-β4 plasmids compared 

with chickens given the pcDNA3-1E vaccine alone. In duodenum, chickens that received 

the pcDNA3-1E vaccine simultaneously with the IL-8 or IL-15 genes had significantly 

increased CD3+ cells compared with vaccination using pcDNA3-1E alone or in 

combination with the other cytokine genes tested. These results indicate that the type and 

the dose of cytokine genes co-injected with a DNA vaccine influence the quality of the 

local immune response to DNA vaccination against coccidiosis. 

Stimulation of chicken leukocytes and reduction of Salmonella organ invasion 

in neonatal chicken by immunostimulatory CpG-oligodeoxynucleotide.

Haiqi He and Michael H. Kogut, 

USDA, ARS, Southern Plains Agricultural Research Center, College Station, TX77845 

Unmethylated CpG oligodinucleotide, flanked by particular base contents, are known to 

stimulate innate immune responses. Immune stimulatory activities of these CpG 

molecules have been studied mostly in immune cells from mammalian, particularly 

human and mice. Recently, studies have emerged to show similar immune stimulatory 

activities of CpG oligodinucleotide in other species such as chicken and fish. Here, we

present results from our studies on immune stimulatory activities of CpG 

oligodinucleotide in chicken. Our results show that synthetic oligodeoxynucleotides 

(ODN) containing CpG motif (CpG-ODN) activated the avian macrophage cell line and 

induced NO production. The most active CpG-ODN motif for NO induction was 

GTCGTT. Increasing the number of GTCGTT motif in the CpG-ODN significantly 

enhanced the stimulatory effect. This CpG-ODN also stimulated IL-1β gene expression 

in the macrophage HD11 cells. CpG-ODN stimulated inducible NO synthase (iNOS) to 

produce NO through a clathrin-dependent endocytosis and subsequent endosomal 

maturation signaling pathways, involving activation of protein kinase C (PKC), mitogenactivated protein kinases (p38 MAPK and MEK1/2), and transcription factor NF-6B. 

Moreover, we have discovered that CpG-ODN induces degranulation in chicken 

heterophils. This heterophil degranulation activity induced by CpG-ODN was absolutely 

serum-dependent and also receptor mediated. Finally, we evaluated synthetic CpG-ODNs 

for their in vivo protection against organ invasion by Salmonella enteritidis (SE) in 

neonatal chickens. Day-old chickens were i.p. injected with synthetic CpG-ODN and then 

orally challenged with live SE. Organs (liver and spleen) were removed from

experimental chickens 24 hours after the challenge and cultured for SE. The results 

showed a significant reduction of organ invasion by SE (p<0.05) in chickens pretreated 

with CpG-ODN containing the immunostimulatory GTCGTT motif. 

2003 ARS Immunology Research Worskshop 41 
Regulation of memory CD8 T cell generation by cytokines

M. Suresh 

University of Wisconsin, School of Veterinary Medicine, Pathobiological Sciences, 

Madison, WI 

http://www.pathology.wisc.edu/gradprogram/trainers/msuresh.html

CD8 T cells play a critical role in defense against viral, intracellular bacterial, and 

protozoon infections. Therefore, vaccines against these viruses need to engender potent 

CD8 T cell memory. Although generation of T cell memory is the basis of vaccinations, 

the mechanism(s) of induction of CD8 T cell memory are not well understood. Typically, 

CD8 T cell responses can be divided into three distinct phases: (1) the expansion phase 

when naive T cells undergo antigen-driven proliferation and differentiation into effector 

cells, (2) the contraction phase when 90-95% of the expanded T cells are eliminated, and 

(3) the memory phase when the remaining ~5% of the expanded CD8 T cells survive for 

extended periods as memory T cells. Upon infection, these memory CD8 T cells respond 

expeditiously and clear the infection by rapidly differentiating into effector T cells. The 

accelerated response that typifies immune memory can be attributed to both quantitative 

and qualitative changes in CD8 T cells: (1) Firstly, in an immune animal, there are more 

antigen-specific T cells, as compared to an immunologically naive animal; (2) Secondly, 

in striking contrast to naive T cells, the memory CD8 T cells in immune animals are 

hyper reactive to antigenic stimulation and capable of efficient trafficking into peripheral 

tissues. The number of memory CD8 T cells generated is dependent upon the extent of 

expansion and the magnitude of contraction during the T cell response. Since protective 

immunity is dependent upon the induction of a threshold number of memory CD8 T cells, 

it is critical to gain a thorough understanding of the mechanisms that regulate the 

expansion and contraction phases of the T cell response. Our studies using the wellcharacterized lymphocytic choriomeningitis virus (LCMV) model in mice have shown 

that signaling via IFNγ receptor (IFNγR) and TNF receptors (TNFR) play important roles 

in limiting the number of antigen-specific memory CD8 T cells that survive the 

contraction phase of the T cell response. In mice, loss of IFNγR reduced the expansion 

of LCMV-specific CD8 T cells, but remarkably ablated the contraction phase of the CD8 

T cell response; IFNγR-deficient mice contained substantially more virus-specific 

memory CD8 T cells, as compared to control wild type mice. Loss of either TNF or TNF 

receptors did not affect the expansion phase of the CD8 T cell response to LCMV, but 

enhanced the survival of LCMV-specific memory CD8 T cells in both lymphoid and nonlymphoid organs. These findings have implications in rational vaccine design and therapy 

of immune-mediated diseases. 

2003 ARS Immunology Research Worskshop 42 
Poster Sessions 

December 2, 2003 

The first author listed presented the poster unless another name is listed in bold

Posters are numbered as indicated. 

Innate Immunity 

Analysis of swine skin-derived dendritic cells in the induction of innate 

immune responses to Foot-and-mouth disease virus (FMDV)

Elida M. Bautista, Geoffrey S. Ferman, Douglas Gregg, Mario C. Brum, Marvin 

Grubman and William T. Golde. 

USDA, ARS, Plum Island Animal Disease Center, ARS, USDA, Greenport, NY 11944. 

See abstract, on page 20. 

The inhibition of nitric oxide production in chicken phagocytes. 

Tawni L. Crippen, Haiqi He and Michael H. Kogut 

USDA-ARS, Southern Plains Area Research Center, College Station,TX 77845. 

The production of reactive nitrogen, nitric oxide (NO), has previously been demonstrated 

to be a major mechanism by which the innate immune system defends against microbial 

invasion. The purpose of this study was to characterize components of the signal 

transduction pathway induced by bacterial stimulation of NO for antimicrobial defense 

by chicken phagocytes. We quantified the production of nitrite by chicken macrophages 

(HD11 cells) which had been exposed to specific signal transduction inhibitors prior to 

stimulation by formalin-fixed Enterococcus gallunarium (EG) or Klebsiella pneumoniae

(KP). The induction of many antimicrobial peptide genes is known to be regulated by 

factors of the NF-6B family in a toll-like-receptor dependent or independent manner. 

NF-6B is in turn regulated by I6B kinase. We found that NO production induced by both 

EG and KP was reduced in a dose dependent manner by specific inhibitors of NF-6B, 

I6B and JAK 3. 

2003 ARS Immunology Research Worskshop 43 
Oligodeoxynucleotides (ODN)-induced activation of macrophage-mediated 

innate immunity against intracellular pathogens, Eimeria and Salmonella,

and the development of novel immunomodulatory strategies using embryo 

injection of CpG-ODNs in poultry.

Dalloul1,2Rami, Lillehoj1 Hyun S., Xie

2

 Hang, Heckert2 Robert A., Babu3 Uma, 

Raybourne3

 Richard, and Klinman4 Dennis M. 

1

USDA, ARS, Animal Parasitic Diseases Laboratory, ANRI, Beltsville, MD, 2VA-MD 

Regional College of Veterinary Medicine, University of Maryland, College Park, MD, 

3

Center for Food Safety & Applied Nutrition, US FDA, Laurel, MD, 4Section of 

Retroviral Immunology, Center for Biologics Evaluation & Research, US FDA, 

Bethesda, MD 

See abstract on page 20.

Toll-like receptor and acute phase cytokine expression in genetically selected 

lines of layers following an LPS challenge

S. D. Eicher and H. W. Cheng 

USDA, ARS, Livestock Behavior Research Unit, West Lafayette, IN 

A multi-disciplinary approach is essential to ascertain the well-being of farm animals. 

The Livestock Behavior Research Unit uses behavioral, physiological, and 

immunological measures to answer well-being questions for poultry, swine, and dairy 

cattle. The innate immune system is frequently the first immune responder during many 

stressors and the resulting acute phase cytokines affect behavior and learning. Toll-like 

receptors are the pathogen recognition molecules that initiate the cascade that increases 

the acute phase cytokines. The effect of pathogens and stressors on the toll-like receptors 

of farm animals is not known. A study has been completed that examines the effect of 

lipopolysaccharide (LPS) on RNA toll-like receptor expression in chickens. The 

objective of this study was to determine expression of TLR2, TLR4, and two acute phase 

cytokines (interleukin-1 and tumor necrosis factor-") following an LPS challenge of 

two genetic lines of layers divergently selected for aggression (aggressive, MBB and 

passive, KGB)and one commercial genetic line (COMM). The 3 lines of layers were 

given LPS (5.0 mg/kg of body weight), then organs (heart, liver, and spleen) were 

weighed and lung, liver, and spleen tissues were collected at 0, 24, and 72 hours 

following a single LPS i.v. injection. RNA was extracted from the tissues and subjected 

to real-time RT-PCR. Heart and spleen weights had genetic line (P < 0.01) and time

effects (P < 0.05 and 0.01, for heart and spleen respectively). Liver weights were 

different for genetic line (P < 0.01) but not time. Liver IL-1 and TLR2 had time effects (P 

< 0.01) but were not different among treatments. Liver TNF and TLR4 were not 

different over time or among treatments. Lung IL-1, TLR2, TLR4, and TNF were 

different over time (P < 0.05 for TLR4 and P < 0.01 for IL-1, TLR2, and TNF) but not 

among treatments. Spleen IL-1, TLR2, TLR4, and TNF were different over time but not 

among treatments (P < 0.05 for IL-1, P < 0.01 for TLR2, TLR4, and TNF). No treatment 

by time interactions were detected for any of these variables (P > 0.05). The only effects 

of genetic line were liver and spleen weight. Toll-like receptors 2 and 4 and cytokine 

changes were not influenced by genetic lines, only by time. Toll-like receptor expression 

was dissimilar for the two receptors that we investigated. Toll-like receptor 2 had effects 

2003 ARS Immunology Research Worskshop 44 
of time in all three tissues, while TLR4 was influenced by time only in lung and spleen. 

Toll-like receptor 2 increase expression in the liver and spleen by 24 h, but did not 

increase in the lung until 72 h post-LPS injection. Liver TLR2 remained elevated 

through the 72 h sample. Therefore, this genetic selection did not alter RNA expression 

of these innate immune receptors and cytokines. Since toll-like receptors recognize heatshock proteins, further exploration of toll-like receptor expression during heat stress in 

chickens is in progress. 

Toll-like receptor and acute phase cytokine expression in neonatal dairy calves

S. D. Eicher1, T. A. Johnson2, K. A. McMunn1, and T. R. Johnson2 

1

USDA, ARS, Livestock Behavior Research Unit, West Lafayette, IN and 2Purdue 

University, West Lafayette, IN. 

See abstract on page 18.

Spleen-dependent innate immunity in calves to Babesia bovis infection is 

characterized as a type-1 response involving the proliferation of NK-like cells

W.L. Goff*1

, W.C. Johnson1, G.M. Barrington2, W. C. Davis3 AND D.P. Knowles1

1

USDA, ARS, Animal Disease Research Unit, Pullman, WA. 99164-6630, 2Department 

of Clinical Medicine and Surgery, and 3

Department of Microbiology and Pathology, 

Washington State University, Pullman, WA 99164 

Innate immunity is important for control of primary infections and for induction of 

acquired responses. Young calves possess a strong spleen-dependent innate immunity to 

the intraerythrocytic protozoan, Babesia bovis. Prominent cellular components in innate 

immunity are dendritic cells, macrophages, and NK-cells. The study reported here 

demonstrates the importance of a type-1 response involving the early induction of IL-12 

and IFN-γ and proliferation of NK-like cells in the spleen of calves during a primary ticktransmitted B. bovis infection. Unlike gdT-cells, CD3-

, CD2+, CD8+ NK-like cells 

proliferated in the spleen, increasing almost three-fold over pre-infection levels. A model 

and hypothesis are presented for macrophage and/or dendritic cell participation with NK 

cells in the induction of a type-1 protective response against this parasite. 

Interferon-( upregulates the expression of proinflammatory and Th1 

cytokine mrna in chicken heterophils during receptor-mediated phagocytosis 

of opsonized and nonopsonized Salmonella enteritidis.

Michael Kogut, Lisa Rothwell, Pete Kaiser 

USDA, ARS, Southern Plains Area Research Center, and Institute for Animal Health, 

College Station, TX 77845 USA and Compton, Berkshire, UK

The immediate response to invasive pathogens, clearance via the inflammatory response, 

and activation of the appropriate acquired responses are all coordinated and orchestrated 

by the innate host defenses. Recognition of microbes is accompanied by the induction of

multiple cell processes including the production of pro- and anti-inflammatory cytokines. 

Polymorphonuclear leukocytes (PMNs) are vital cellular components of innate immunity, 

and function by killing pathogenic microbes following phagocytosis. The primary PMN in 

poultry is the heterophil. Priming is the potentiation of the phagocyte activation process by 

2003 ARS Immunology Research Worskshop 45 
previous exposure to a priming agent. IFN-( is a pleiotrophic cytokine involved in basically 

all phases of immune and inflammatory responses that has been shown to prime heterophil

functional activities. In the present experiments, using real-time quantitative RT-PCR, we

evaluated the role of rChIFN-( as a priming mediator to control heterophil responses at the

level of gene transcription and expression of the mRNA for proinflammatory (IL-1b, IL-6, 

IL-8) and Th1 (IL-18 and IFN-() cytokine genes following stimulation with phagocytosis 

agonists opsonized and nonopsonized Salmonella enteritidis. RChIFN-( primed the 

heterophils for an increase in transcription of pro-inflammatory cytokines induced by 

phagocytic agonists, but also upregulated expression of Th1 cytokine (IL-18 and IFN-() 

mRNA. Although rChIFN-( priming modulated the expression of cytokine mRNA in 

heterophils stimulated by different phagocytic agonists, the rChIFN-( by itself did not 

directly induce gene expression of either the proinflammatory or Th1 cytokines. The

enhanced expression of cytokine mRNA does not appear to be differentially expressed 

depending on the receptor activated during phagocytosis. The results from the present 

experiments suggest that rChIFN-( may play a significant role in avian innate immunity 

against Salmonella infection and may offer an adjunct use in the prevention and treatment of 

salmonellae infections in newly hatched chickens. 

Recombinant bovine soluble CD14 reduces fatality of endotoxin challenged

mice and reduces severity of intramammary infection by Escherichia coli in 

dairy cows.

Max J. Paape, Jai-Wei Lee and Xin Zhao 

USDA, ARS, Beltsville, Maryland and McGill University, Ste-Anne-de-Bellevue, 

Quebec, Canada 

Three million cases of clinical mastitis caused by Gram-negative bacteria occur every 

year in the U.S. A significant number of these cases will result in acute endotoxin shock 

and death. Three hundred thousand dairy cows are removed from herds or die annually 

because of acute coliform mastitis. Standard therapy for treating Gram-negative bacterial 

sepsis and shock, including administration of potent antibiotics, aggressive fluid 

resuscitation, and metabolic support, has not been successful in reducing mortality. 

Soluble CD14 (sCD14) neutralizes endotoxin and prevents it from binding and activating 

macorphages. Further, LPS/CD14 complexes bind to epithelial cells and results in rapid 

recruitment of neutrophils to the infection site. Novel reagents such as host defense 

proteins like sCD14 have a role as a potential prophylactic against serious Gram-negative 

bacterial infections. In the present study, the full length and the truncated forms of 

bovine sCD14 were cloned into a transfer vector and expressed by recombinant 

baculovirus that infect insect cell line sf-9. The protective effect of the recombinant 

bovine sCD14 (rbosCD14) was evaluated in mice and lactating dairy cows. Eighty–one 

female mice were randomly assigned to two groups and injected intraperitoneally with 

either LPS (8 μg/g BW, n = 41) or LPS plus rbosCD14 (6.8 μg/g BW, n = 40). Survival 

rate for LPS and LPS plus rbosCD14 injected mice at 24 h was 30 and 72%. Two 

mammary quarters of each of 9 cows were challenged with either 50 CFU of Escherichia 

coli plus saline or 50 CFU of E. coli plus 100 μg rbos CD14. Quarters challenged with E. 

coli plus rbosCD14 had a more rapid recruitment of neutrophils, which was accompanied 

by a faster clearance of bacteria and reduced clinical symptoms than challenged quarters 

2003 ARS Immunology Research Worskshop 46 
injected with saline. At 24 h after bacterial challenge, only two (22%) of nine quarters 

challenged with E. coli and injected with rbosCD14 remained infected. On nine 

challenged quarters injected with saline, six (67%) remained infected. Results indicated 

that rbosCD14 was able to decrease the fatality of LPS challenged mice and reduced 

incidence of infection in dairy cows following experimental challenge with E. coli. Use 

of rbosCD14 in the treatment of and prevention of acute coliform mastitis has the 

potential of saving US dairymen 1.4 billion dollars annually. 

Immune Regulation 

Systemic cytokine and antibody responses following vesicular stomatitis virus 

infection of naïve and vaccinated cattle.

José Barrera 1, 2, William T. Golde1, D. Mark Estes2 and Luis L. Rodriguez1

1

USDA, ARS, Plum Island Animal Disease Center, Greenport, NY, 11944,2 University of 

Missouri. College of Veterinary Medicine. Deptartment of Veterinary Pathobiology, 

Columbia, MI, 65211. 

Vesicular stomatitis virus (VSV) causes disease of varying severity in cattle, swine, and 

horses. The mechanisms mediating protection from clinical disease remain unknown. 

This study was carried out to gain a better understanding of the systemic T-helper cell 

balance mediating the immune response against VSV in cattle. We compared two cattle 

vaccinated with a commercial oil adjuvanted bivalent vaccine and four naïve animals, all 

challenged with a virulent strain of VSV New Jersey (VSNJV). Vaccinated animals 

developed high titers of neutralizing antibodies against VSNJV and VSV Indiana 

serotypes (VSIV) (range from 2.5 to 4.6 log) and showed no disease following challenge. 

In contrast, non-vaccinated naïve cattle developed fever and lesions. VSV was recovered 

from epithelial tissue and from esophageous-pharyngeal and vesicular fluids. VSNJV - 

specific IgM antibodies were detected in naïve infected cattle at 4 days post-infection 

(dpi) and class switch to IgG1 and IgG2 occurred by 14 dpi and 14 days post-vaccination 

(dpv) in naïve and vaccinated cattle respectively. Titers of IgG2 isotype were higher than 

those of IgG1 isotype (IgG2/IgG1 ratio ranged from 1.2 to 6.5) both in naïve and 

vaccinated animals. IFN-α, IFN-β, IFN-γ, TNF-α, IL-1b, IL-2, IL-12p40, IL-4, IL-10 and 

IL-13 were measured by real-time reverse transcription-PCR (RT-PCR) in fresh 

peripheral blood mononuclear cells (PBMC) or PBMC stimulated with VSNJVantigen 

included. Transcripts for IFN-α, IFN-γ, and IL-12p40 were more abundant than those for 

IL-4, IL-10, and IL-13 in PBMC cultures -stimulated with VSNJV. Taken together, these 

results suggest that VSV antigen induces a T-helper Type 1 response in both infected and 

vaccinated cows. We propose that analysis of cellular responses at the sites of lesion and 

in draining lymph nodes, would give a more accurate picture of the events associated 

with development of immunity to this localized infection in domestic animals. 

Mycoplasma gallisepticum: competitive exclusion or immune base

Stephanie D. Collier 

USDA, ARS, Mid-South Area, Poultry Research Unit, Mississippi State, MS 

2003 ARS Immunology Research Worskshop 47 
http://msa.ars.usda.gov/ms/msstate/csrl/collier.htm

Much confusion exists concerning the mechanism(s) by which the available Mycoplasma

gallisepticum (MG) vaccines confer protection in layer chickens. One of the major goals 

of this laboratory is to discern the role of competitive exclusion and/or immune based 

mechanisms following vaccination with MG. The parameters to be examined in the 

present study include T-cell subpopulations (CD4 and CD8) and cytokines levels (IFNgamma, IL-6 and IL-8) immediately before and following MG vaccination via flow 

cytometry and real-time qRT-PCR. 

Development of specific immunoglobulin IgGa and IgGb antibodies correlate 

with control of parasitemia in Babesia equi infection: IgG isotype responses in 

B. equi infection.

Cristina W. Da Cunha1, 2, Travis C. McGuire1, Lowell S. Kappmeyer3, Stephen A. 

Hines1

, Alicia M. Lopez1, Odir A. Dellagostin2, AND Donald P. Knowles1, 3 *

1

Department of Veterinary Microbiology and Pathology, Washington State University, 

Pullman, WA, 99164-7040 ; 2

Center for Biotechnology, Federal University of Pelotas, 

RS, Brazil, 99010-900; 3

USDA, ARS, Animal Disease Research Unit, Pullman, WA, 

99164-6630 

Following infection with Babesia equi horses often survive acute disease but remain lifelong carriers. Acquired immune responses are necessary to control replication while 

adaptive immune responses are not required for parasite-mediated erythrocyte lysis. 

Following lysis of infected erythrocytes, parasites are accessible to antibody and 

associated mechanisms of antibody-mediated control. To determine if IgG isotypes to B. 

equi erythrocyte stage antigens correlate with parasite control, antibody isotypes were 

measured following infection by parenteral inoculation or by the tick vector Boophilus 

microplus. Isotypes IgGa, IgGb, and IgG(T) were titrated in B. equi infected horse serum

by ELISA using B. equi erythrocyte stage extract as antigen and monoclonal antibodies 

specific for each equine subisotype. IgGa and IgGb antibodies to B. equi developed 

during acute infection, but IgG(T) antibodies were detected only after resolution of acute 

parasitemia. During chronic infection, antibodies of all three isotypes were present at 

titers of 103 to 104

 in naturally and experimentally infected horses. To test whether 

immunization induced IgGa and IgGb antibodies and not IgG(T) antibodies similar to 

those during acute infection with B. equi, recombinant erythrocyte stage surface protein

EMA-1 in the adjuvant saponin was tested. Four immunized horses developed IgGa and 

IgGb antibodies to B. equi, but no measurable IgG(T) antibodies. These results 

demonstrated that development of IgGa and IgGb antibodies to B. equi erythrocyte stage 

antigens correlated with control of B. equi parasitemia, and that this IgG isotype profile 

was induced by subunit immunization. 

Analysis of lymphocytes isolated from white-tailed deer (Odocoileus 

virginianus) fawns 

Sasha J. Fach1

; W.Ray Waters2, Mitchell V. Palmer

2

; William C. Davis3;Randy E. 

Sacco2

2003 ARS Immunology Research Worskshop 48 
1

 Immunobiology Graduate Program, Iowa State University, Ames, IA 50010; 2 USDA, 

ARS, National Animal Disease Center, Ames, IA 50010; 3 Department of Veterinary 

Microbiology and Pathology, College of Veterinary Medicine, Washington State 

University, Pullman, WA 99164 

Peripheral blood mononuclear cells (PBMC) were isolated from ten female white-tailed 

deer fawns at 48 hours of age and every two weeks there after until the fawns were three 

months of age. Bone marrow aspirates and peripheral blood samples were taken on the 

same fawns at ten months of age (yearlings). In a separate experiment, mesenteric lymph 

node (MLN), spleen, thymus, bone marrow (BM) and peripheral blood (PB) were 

collected from euthanized female fetuses at approximately 190 days gestation (n=4) and 

from fawns at 24 hour (n=2), two weeks (n=2) and four weeks (n=2) of age. Mononuclear 

cells were isolated from pooled, homogenized tissues and individual peripheral blood 

samples at each of the four time points. Lymphocytes were phenotyped to examine the 

expression of specific surface receptors as the fawns aged. Three-color flow cytometric 

analysis of leukocytes was performed using a panel of monoclonal antibodies (mAb). 

Adult deer were also phenotyped using the same monoclonal antibody panel. Included in 

the panel were mAbs recognizing WC1, the chain and chains, CD4, CD8, CD62L, 

CD44, CD21, IL-2R, MHC Class II, B-B1, B-B2 and B-B4. Analysis revealed dynamic 

changes in the expression of specific surface receptors associated with development and 

maturation of lymphocyte subpopulations. WC1+

 gamma delta T cells were predominant 

in the PB of fetal and neonatal fawns and decreased with age. In contrast, percentages of 

CD4+

 and CD8+ populations were observed to increase over time in the PB. T cell surface 

antigen expression in the PB and tissues was consistent with observations made in other 

ruminants. B cell surface IgM expression was heterogeneous at two days, but became 

more discrete as the fawns matured. Interestingly, B-B2, a putative B cell lineage marker 

expressed on lymphocytes in the PB at 24-48 hours was down modulated by two weeks. 

However, expression of another B cell lineage marker, B-B4, was consistently expressed 

throughout the fawns’ development in the PB, MLN, and spleen. Mononuclear cells 

isolated from bone marrow aspirates revealed phenotypically distinct expression of 

surface receptors as compared to PBMCs. Mononuclear cells co-expressing B-B2 and 

surface IgM were a unique population found in the bone marrow and not in the PB. The 

sIgM+

B-B2+ population was observed in the fetal spleen and MLN. B-B4 was expressed 

on a small percentage of bone marrow mononuclear cells. Proliferative responses of the 

isolated PBMC’s to pokeweed mitogen (PWM), Concanavalin A (ConA) and 

Mannheimia (Pasteurella) haemolytica LPS were also examined. Cells isolated from

fawns proliferated in response to PWM and ConA, but not in response to LPS 

stimulation. Data obtained in the present study provides baseline information regarding 

lymphocyte subpopulations in white-tailed deer fawns. 

Effects of the mammary gland on the composition and function of peripheral 

blood mononuclear leukocyte populations in periparturient dairy cows. 

J.P. Goff, K. Kimura, B.J. Nonnecke 

Periparturient Disease of Cattle Research Unit 

USDA, ARS, National Animal Disease Center, Ames, IA 50010 

See abstract on page 25. 

2003 ARS Immunology Research Worskshop 49 
Comparison of adhesion molecules on milk and blood lymphocytes from 

periparturient dairy cattle.

James A. Harp*, Theresa E. Rahner, and Jesse P. Goff. 

USDA, ARS, Periparturient Diseases of Cattle Research Unit, National Animal Disease 

Center, Ames, IA 50010. 

Fifteen Holstein dairy cows were monitored for expression of adhesion molecules on 

lymphocytes in blood and milk at parturition and at intervals up to 21 days post partum. 

Using flow cytometric analysis, we examined expression of CD62L, CD11a, CD44, and 

α4β7 on T (CD4+, CD8+, γδ+) and B lymphocytes. CD62L was expressed on 20-70% of 

T cell subsets in blood and 35-80% in milk from normal cows. CD62L expression on B 

cells was 20-25% in blood and 60-80% in milk. CD11a was found on 10-25% of T cells 

in blood and 35-70% in milk. CD11a was found on 10-15% of B cells in blood and 45-

75% in milk. α4β7 was expressed on 5-10% of T cells in blood and 10-35% in milk. α4β7

was expressed on 5-10% of B cells in blood and 20-40% in milk. CD44 was found on 5-

15% of T cells in blood and 20-55% in milk. CD44 was found on 5-10% of B cells in 

blood and 30-55% in milk. There were consistently more adhesion molecules, at a 

statistically significant level, on milk lymphocytes compared with blood lymphocytes. 

These differences may reflect increased recruitment of lymphocytes into the bovine 

mammary gland, which could then modulate the inflammatory response to pathogens 

during infection. Since CD62L and α4β7 are peripheral and mucosal homing receptors, 

respectively, these data suggest increased recruitment of both pools of lymphocytes into 

the bovine mammary gland. 

Evaluation of the chicken crop (ingluvies) as a model system for following 

mucosal immunity in chickens.

Peter S. Holt, Ph.D. 

USDA, ARS, Southeast Poultry Research Laboratory, 934 College Station Road, Athens, 

GA 30605. 

The crop (ingluvies) is an enlargement or out pouching of the esophagus proximal to the 

proventriculus, or glandular stomach and functions primarily in a food storage role for 

avian species. While recent work has shown the crop can be colonized with pathogens 

such as Salmonella, little information is available regarding the presence of any immune 

response within this organ. During a recent study investigating mucosal immunity to 

Salmonella enteritidis (SE) in the alimentary tract of White Leghorn chickens, we 

discovered the presence of substantial titers of IgA specific for SE in samples from crops 

removed from SE-infected, but not from noninfected, birds. Histological examination of 

the crop tissues found a paucity of lymphocytes in noninfected birds but lymphocyte 

aggregates were detected in the crop mucosa by 2 weeks post infection which increased 

in size and number thereafter. Because of the easy access to this organ orally, a crop 

lavage system was developed where a narrow diameter plastic tubing attached to a 

syringe was inserted down the esophagus into the crop. A flush solution was 

administered into the crop and immediately aspirated back into the syringe, with minimal 

discomfort to the bird and allowing multiple samplings of the same live birds over time. 

2003 ARS Immunology Research Worskshop 50 
While specific IgA responses were the most robust, high levels of SE-specific IgG were 

also detected. Comparing IgA titers in crop lavage samples vs intestinal flushes from

individual SE-infected birds, a very good correlation between titers of the two sample 

types was obtained, indicating that crop immunity may be used as an easy model for 

following mucosal immunity in a bird following infection. Studies are currently 

underway to determine crop immune responses in chickens infected with other 

pathogens, the prevalence of lymphoid tissues in crops from different avian species, the 

phenotypes of lymphocyte populations that develop in the chicken crop post infection and 

the importance of mucosal invasion in driving the development of this lymphoid tissue. 

Neutralization of bluetongue virus in vertebrate and invertebrate cells.

James O. Mecham

USDA, ARS, Arthropod-borne Animal Diseases Research Laboratory, Laramie, WY 

Bluetongue virus (BTV) is an orbivirus that infects sheep, cattle and wild ruminants. It is 

transmitted to susceptible vertebrate hosts by biting midges in the genus Culicoides. 

Following infection, the vertebrate host produces neutralizing antibodies that are 

responsible for virus clearance and conferring immunity to subsequent virus challenge. 

These neutralizing antibodies are directed against the virus outer capsid, which is also

responsible for initiating infection in the vertebrate host. Infection of the insect vector is 

believed to be mediated primarily by the inner capsid, which is not the target of 

neutralizing antibody. Therefore, virus that is neutralized for infection of vertebrate cells 

may still be infectious for invertebrate cells. This may have important implications in the 

natural epidemiology of BTV infections. To investigate this further, the mechanisms of 

neutralization of BTV in both vertebrate and invertebrate cells was investigated. 

Antibodies that neutralize BTV infection in vertebrate cells failed to neutralize viral 

infection in a Culicoides cell line. However, non-neutralizing antibodies that react with a 

virus core protein inhibited infection and/or replication of BTV in the Culicoides cell 

line. Since this virus core protein is highly conserved between BTV serotypes, and 

appears to play an important role in the initiation of BTV infection in insect cells, it may 

represent a novel target for control of infection caused by these viruses. 

2003 ARS Immunology Research Worskshop 51 
Comparison of the adaptive immune response in neonatal and young adult dairy 

cattle utilizing a Mycobacterium bovis BCG sensitization/PPDb challenge model.

B. J. Nonnecke1

, M. R. Foote2, B. L. Miller3, M. A. Fowler3, W. R. Waters4

1

USDA, ARS, Periparturient Diseases of Cattle Research Unit, National Animal Disease 

Center, Ames, IA, 2

Iowa State University, Ames, IA, 3Land O’Lakes, Inc., Webster City, 

IA and 4

USDA, ARS, Bacterial Diseases of Livestock Research Unit, National Animal 

Disease Center, Ames, IA. 

Acquisition and modulation of immune competency in calves during the neonatal and 

postnatal period are not well described. The purpose of this study was to characterize 

age-related changes in the functional capacities of peripheral blood mononuclear cell 

(PBMC) and neutrophil populations from antigen-sensitized calves. Milk replacer-fed 

Holstein bull calves were nonvaccinated (n = 6, vaccination controls) or vaccinated subQ 

(n = 6) with an attenuated strain of Mycobacterium bovis [bacille Calmette Guerin 

(BCG)] at 1 and 7 wk of age. Yearling heifers (n=4, adult controls) were vaccinated and 

sampled concurrently with the calves. Functional capacities of PBMC (i.e. mitogen and 

antigen-induced DNA-synthesis, T cell subset proliferation, cytokine and NO production, 

IgG secretion) and neutrophil (iodination and cytochrome C reduction) populations 

collected at 0 (pre-vaccination), 2, 5, 6 (boosted), 7, 8 and 11 wk were evaluated in vitro. 

The phenotype of PBMC populations was characterized by flow cytometry. The 

responsiveness of PBMC from nonvaccinated and vaccinated calves to polyclonal 

stimulation frequently was lower than adult PBMC. Responses of PBMC from

vaccinated calves to recall antigen (PPDb) were evident at >2 wk (3 wk of age) after 

primary vaccination. With the exception of antigen-induced DNA synthesis, most 

functions (T cell subset proliferation, TNF-α, IFN-γ and NO secretion) of PBMC from 

vaccinated calves were comparable to or exceeded those of vaccinated adults. Changes in 

cervical skin-fold thickness after intradermal administration of PPD (wk 11) were 

significant and comparable in vaccinated calves and adults confirming the development 

of a robust cell-mediated immune response in vaccinated calves. In contrast, antigenspecific IgG responses (i.e. PPD-specific IgG in sera and supernatants from PPDstimulated cultures) of vaccinated calves were minimal or nonexistent relative to adult 

responses suggesting the contribution of the B cell to the immune response is limited in 

young cattle. Nonvaccinated calves did not respond to recall antigen in vivo (i.e. 

cutaneous delayed-type hypersensitivity) or in vitro. Neutrophil functions were affected 

by age but not vaccination. 

2003 ARS Immunology Research Worskshop 52 
Molecular characterization of a putative Tumor Necrosis Factor superfamily 

13b (TNFSF 13b) homologue from rainbow trout (Oncorhynchus mykiss).

Gregory D. Wiens*, Katherine Hovatter, Yniv Palti, Carid Rexroad III, Timothy Welch. 

USDA/ARS National Center for Cool and Cold Water Aquaculture, 11871 Leetown 

Road, Kearneysville, WV 25430 

Little is known about the immune system function of economically important cool and 

cold water fish species. Such knowledge is important for developing successful vaccines 

and disease resistant fish. In mammals, members of the tumor necrosis factor 

superfamily (TNFSF) play important roles in both innate and adaptive immune responses. 

In order to determine which TNFSF members are conserved and expressed in rainbow 

trout, we have searched a computer database of ~50,000 expressed sequence tags 

generated at the National Center for Cool and Cold Water Aquaculture. We have 

identified 17 cDNAs that have homology to known TNFSF ligands or receptors. Here, 

we describe one cDNA, 1RT126I22, which has homology to TNFSF 13b. TNFSF 13b is 

also known as B-cell activating factor (BAFF) or B-lymphocyte stimulating factor 

(BLYS). In mice and humans, TNFSF 13b is produced by myeloid cells and is critical 

for mature B cell survival and antibody production. The rainbow trout cDNA, 

1RT126I22, encodes a predicted 141 amino acid protein, which shares 54% amino acid 

identity with human TNFSF13b, 51% identity with mouse TNFSF13b, and 54% identity 

with chicken TNFSF13b. The rainbow trout TNFSF 13b shares 100% identity with a 99 

amino acid protein predicted from a truncated Atlantic salmon cDNA, and 78% identify 

with puffer fish TNFSF13b protein predicted from genomic DNA sequence. The 

rainbow trout TNFSF 13b is unusual as it does not possess a transmembrane domain or a 

furin-type protease cleavage site characteristic of the human, mouse, and chicken TNFSF 

13b molecules. Examination of rainbow trout TNFSF 13b gene expression during 

vaccination and bacterial challenge is underway. To our knowledge, this is the first 

identification and characterization of fish TNFSF 13b homologues. 

Immunogenetics 

Expression of specific MHC Class II DRB1 alleles associates with sheep 

persistently infected with ovine progressive pneumonia virus (OPPV).

L.M. Herrmann1

, W.P. Cheevers2, D.P. Knowles

1,2

1

USDA, ARS, Animal Disease Research Unit, Pullman, WA 99164-6630. 2 Department 

of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 

99164-7030. 

See abstract on page 28.

2003 ARS Immunology Research Worskshop 53 
Marek's disease and the MHC: How viruses subvert the immune system for 

their advantage.

H. Hunt, H. Cheng, and H. Zhang 

USDA, ARS, Avian Diseases and Oncology Laboratory, East Lansing, MI 

See abstract on page 27.

Physical and genetic mapping of the rainbow trout major histocompatibility 

regions: evidence for duplication of the class I region

Ruth Phillips

1

, Ana Zimmerman2, Marc Noakes1, Yniv Palti3, Matt Morash1, Lisa 

Eiben4

, Sandra S. Ristow2, Gary H. Thorgaard2 and John D. Hansen4. 1

School of Biological Sciences, Washington State University, 14204 NE Salmon Creek 

Ave. Vancouver, WA 98686-9600 

2

School of Biological Sciences, P.O. Box 644236, Washington State University, Pullman, 

WA 99164-4236 

3

USDA, ARS-NCCCWA, 11876 Leetown Road, Kearneysville, WV 25430 

4

Center of Marine Biotechnology, University of MD Biotechnology Institutes, 701 E.

Pratt St, Baltimore, MD 21202. 

Previous segregation analysis in trout suggested that the class I and II regions reside on 

completely different chromosomes. To learn more about MHC genomics in trout, we 

have isolated BAC clones harboring class IA and IB loci, a single BAC clone containing 

an MH class II gene (DAB) as well as BAC clones containing the ABCB2 gene. Upon 

PCR and sequence confirmation, BAC clones were labeled and used as probes for in situ

hybridization on rainbow trout metaphase chromosomes for determination of the physical 

locations of the trout MH regions. Finally, SNPs, RFLPs and micro-satellites found 

within the BAC clones allowed for these regions to be assigned to specific linkage groups 

on the OSUxHotcreek and OSUxArlee genetic linkage maps. Our data demonstrates that 

the trout MH regions are located on at least four different chromosomes and the 

corresponding linkage groups while also providing direct evidence for the partial 

duplication of the MH class I region in trout. 

2003 ARS Immunology Research Worskshop 54 
Immunity and the Pathogenesis of Disease 

A dual role for flice-like inhibitory protein (flip) in mediating both bacterial 

lipopolysaccharide-induced apoptosis and NF-κb activation.

Douglas D. Bannerman

1

, Kristine T. Eiting2, Robert K. Winn2, and John M. Harlan2

1

 USDA, ARS, Immunology and Disease Resistance Laboratory, Beltsville, MD 20705 

and 2

University of Washington School of Medicine, Seattle, WA 98104 

Bacterial lipopolysaccharide (LPS) contributes to much of the vascular 

injury/dysfunction associated with Gram-negative sepsis. We have previously shown 

that inhibition of de novo gene expression with cycloheximide sensitizes vascular 

endothelial cells (EC) to LPS-induced apoptosis and that the onset of apoptosis correlates 

with decreased expression of FLICE-like inhibitory protein (FLIP), an anti-apoptotic 

protein. We now have data that conclusively establish a role for FLIP in protecting EC

against LPS-induced apoptosis. Downregulation of FLIP using anti-sense 

oligonucleotides sensitized EC to direct LPS killing, whereas, overexpression of FLIP 

protected against LPS-induced apoptosis in the presence of cycloheximide. Interestingly, 

FLIP overexpression completely blocked NF-kB activation induced by LPS, but not by 

phorbol ester, suggesting a specific role for FLIP in mediating LPS activation. Together, 

these data support a dual role for FLIP in mediating LPS-induced apoptosis and NF-kB 

activation. 

Gene expression profiling of bovine macrophages in response to Escherichia 

coli O157:H7 lipopolysaccharide

Carol G. Chitko-McKown*, James M. Fox, Laura C. Miller, Michael P. Heaton, James L. 

Bono, James E. Keen, William M. Grosse1

, William W. Laegreid 

USDA, ARS, Roman L. Hruska U.S. Meat Animal Research Center, Clay Center, 

Nebraska 68933-0166 

The aim of this study was to identify changes in bovine macrophage gene expression in 

response to treatment with Escherichia coli 0157:H7 lipopolysaccharide (LPS), utilizing 

a human gene microarray. Bovine cDNA from control and LPS-treated primary 

macrophages hybridized to greater than 5,644 (79.8%) of the non-control gene targets on 

a commercially available microarray containing greater than 7,075 targets (Incyte 

Genomics, St. Louis, MO). Of these target sequences, 44 were differentially expressed 

upon exposure to LPS, including 18 genes not previously reported to exist in cattle. 

These included a pentaxin-related gene, CASP8, TNF-induced genes, interferon-induced 

genes, and inhibitors of apoptosis. Using the human microarray, cDNA from bovine 

LPS-treated and control macrophages consistently hybridized to targets known to be 

expressed constitutively by macrophages, as expected given the predicted cDNA 

sequence homology. That this human system was accurately estimating levels of bovine 

transcripts was further verified by real-time quantitative reverse transcriptase polymerase 

chain reaction (RTQ-PCR) using bovine-specific primers. This first report of bovine2003 ARS Immunology Research Worskshop 55 
human cross-species expression profiling by microarray hybridization demonstrates the 

utility of this technique in bovine gene expression and discovery. 

Evaluation of immune response induced by attenuated and wild-type PRRS virus

J Janutenaite 1, K M Lager2, T Stabel2, B Pesch2 , S LBrockmeier2 1

National Veterinary Laboratory, Vilnius, Lithuania; 2National Animal Disease Center, 

USDA, Agricultural Research Service, P O Box 70, Ames, Iowa 50010, USA 

Introduction and Objectives 

Porcine reproductive and respiratory syndrome virus (PRRSV) modified-live vaccines 

(MLV) have been developed by attenuation in cell culture. Under experimental 

conditions the vaccines are efficacious; however, under field conditions vaccine efficacy 

reports have ranged from excellent to poor. One possible reason for questionable efficacy 

is that MLV may not induce a similar immune response when compared to wild-type 

PRRSV infection. The goal of the study reported here was to compare the cellular 

immune response in pigs following exposure to an attenuated PRRSV or its wild-type 

parent virus. 

Materials and Methods

Two experiments were conducted using 52 4-5 week-old conventionally-raised pigs free 

of PRRSV and PRRSV-specific antibody. PRRSV strain NADC-8 (3rd passage) and 

NADC-8 (251st passage) were wild-type parent virus and attenuated virus, respectively. 

Based on sequence of ORFs 2-7 (3,184 nucleotides) the attenuated virus was 99.4% 

homologous with its parent virus. A different wild-type PRRSV strain, SDSU 73 (3rd

passage) was used in the 4th group in experiment 2. Each experiment had a similar 

design: a sham-inoculated control group (group 1), an attenuated virus-inoculated group 

(group 2) and a wild-type parent virus-inoculated group (group 3). In experiment 2 there 

was a 4th group inoculated with SDSU 73. There were 13, 15, 15 and 8 pigs in groups 1, 

2, 3 and 4, respectively. Pigs were each inoculated intramuscularly with 2 ml of their 

respective virus inoculum (about 2 x 105

 TCID50 virus/ml ) day 0, pigs in group 1 

received 2 ml of sham inoculum. Blood samples were collected on days -7, -4, 0, 3, 6, 9, 

13 and 20 ( 21 for experiment 2). Serum was tested for PRRSV specific antibodies and 

used for PRRS virus isolation. A rapid flow cytometric method (6) and a panel of 

monoclonal antibodies were used to analyze porcine peripheral blood leukocytes by twocolor flow cytometric analysis. Statistical analysis was performed using GraphPad InStat 

software. 

Results and Discussion

Results represent the combined data from the two experiments. Statistically significant

changes (P<0.05) in lymphocyte subpopulations percentage were observed in groups 3 

and 4; whereas no changes were observed in group 2. CD8+ T cells increased on days 6, 

9 and 13 for group 3 and on days 9 and 13 for group 4. CD4+ T cells increased on day 9 

for group 4. CD3+ T cells increased on day 9 for groups 3 and 4. Cells expressing Class 

II marker decreased on day 3 in group 3 and on days 3 and 6 in group 4; Class II cells 

increased in the both groups on days 13 and 20(21). CD45+ cells increased on days 9 and 

13 for groups 3 and 4. B cells decreased on days 3, 6 and 9 for group 3 and on days 3 and 

2003 ARS Immunology Research Worskshop 56 
6 in group 4. Total WBC counts decreased on days 6 and 9 in group 3 and on days 3, 6 

and 9 in group 4. Virus was isolated from all wild-type PRRSV-infected pigs (groups 3 

and 4), but no virus was isolated from the sham or attenuated-virus inoculated pigs 

(groups 1 and 2). All wild-type PRRSV-inoculated pigs and 6 of the 15 attenuated-virus 

inoculated pigs (group 2) developed PRRSV-specific antibody. No sham-inoculated pigs 

developed antibody. Group 2 data were further analyzed by dividing pigs in seropositive 

and seronegative subgroups. No statistical differences in percentage of lymphocyte 

populations were found. 

The percentage lymphocyte values determined in this study for the naïve animals 

represent normal values for conventionally raised pigs and are in agreement with 

previously reported data (6). This agreement of data suggests the methodology used in 

these experiments is reproducible. The peripheral blood cellular immune response 

characterized in the pigs infected with wild-type PRRSV appears similar to what has 

been reported by others. Following experimental infection with wild-type PRRSV there 

may be a transitory decrease in CD4+ and CD8+ T cells within the first few days. This is 

followed by a return of CD4+ cells to normal levels and an elevated level of CD8+ cells 

that may last for at least several weeks (1-5) Collectively, these studies in conjunction 

with the present study suggest T cells play an important role in the PRRSV immune 

response following wild-type virus infection. No significant changes in blood cell 

populations were noted following infection with the attenuated PRRSV (group2) when 

compared to negative control group. Perhaps this observation may account for the lack of 

vaccine efficacy that has been described under field conditions. 

The study reported here indicates that the magnitude of the PRRSV–specific immune 

response may be related to PRRSV virulence. Additional studies are needed to confirm 

this hypothesis and what, if any, relevance this may have on vaccine development. 

References: 

Albina E, et al. 1998. Vet Immunol Immunopathol 61:49-66. 

Meier W, et al. 1999. Proc PRRS & Aujeszky’s Dis Symp pp. 71-74. 

Nielsen J, Botner A. 1997. Vet Microbiol 55:289-294. 

Samson JN, et al. 1999. Proc PRRS & Aujeszky’s Dis Symp pp. 81. 

Shimizu M, et al. 1996. Vet Immunol Immunopathol 50:19-27. 

Stabel T, et al. 2000. J Immunol Methods 245:147-152. 

2003 ARS Immunology Research Worskshop 57 
Decreased neutrophil function as a cause of retained placenta in dairy cattle

Kayoko Kimura, Jesse P. Goff, Marcus E Kehrli, Jr., and Timothy A. Reinhardt 

USDA, ARS, National Animal Disease Center, Periparturient Disease of Cattle Research 

Unit Ames, IA 50010 

It is unclear why some cows fail to expel the placenta following calving. One theory 

suggests the fetal placenta must be recognized as “foreign” tissue and rejected by the 

immune system after parturition to cause expulsion of the placenta. We hypothesized that 

impaired neutrophil function causes retained placenta (RP). We examined the ability of 

neutrophils to recognize fetal cotyledon tissue as assessed by a chemotaxis assay. 

Neutrophil killing ability was also estimated by determining myeloperoxidase activity in 

isolated neutrophils. Blood samples were obtained from 142 periparturient dairy cattle in 

2 herds. Twenty cattle developed RP (14.1 %). Neutrophils isolated from blood of cows 

with RP had significantly lower neutrophil function in both assays prior to calving and 

this impaired function lasted for 1-2 wk after parturition. Addition of antibody directed 

against interleukin-8 (IL-8) to the cotyledon preparation used as a chemoattractant 

inhibited chemotaxis by 41% suggesting one of the chemoattractants present in the 

cotyledon at parturition is IL-8. At calving plasma IL-8 concentration was lower in RP 

cows (51 ± 12 pg/ml) than in cows expelling the placenta normally (134 ± 11 pg/ml). 

From these data, we suggest that neutrophil function is a determining factor for the 

development of RP in dairy cattle. Also, depressed production of IL-8 may be a factor 

affecting neutrophil function in cows developing RP. 

Comparative proteomic analysis of normal and periparturient bovine 

neutrophils demonstrating a reduction of myeloperoxidase during transition.

John D. Lippolis, Jesse P. Goff and Timothy Reinhardt 

USDA, ARS, National Animal Disease Center, Ames IA 50010 

Neutrophils constitute a primary effector of the host immune response to an infectious 

pathogen. Normally low numbers of neutrophils reside in a health mammary gland, 

however, infection and subsequent release of chemoattractants results in the rapid influx 

of activated neutrophils into the site of infection. Periparturient dairy cows exhibit signs 

of immunosuppression that correlate with a higher incidence of mastitis. Furthermore, 

periparturient immunosuppression includes a significant reduction of neutrophil function. 

To better understand the nature of the periparturient immunosuppression, neutrophils 

from dairy cows were isolated greater than 21 days prior to calving (normal function) and 

shortly after calving (immunosuppressed) and compared using differential mass 

spectrometry. Proteomes of the immunosuppressed and normal neutrophils were labeled 

with isotope-coded affinity tags (ICAT), digested and subjected to nano-flow LC-MSMS 

analysis. Neutrophil proteins were identified and quantitated, interestingly 

myeloperoxidase was shown to be reduced three-fold around parturition whereas 

cathelicidin 1 and beta defensin were unaffected. Myeloperoxidase is a strongly 

bactericidal neutrophil enzyme that generates reactive oxidants and is considered an 

important component of neutrophil’s ability to destroy bacteria. 

2003 ARS Immunology Research Worskshop 58 
The effects of Pasteurella multocida toxin (PMT) on early murine B-lineage cells

Crystal L. Loving and Randy E. Sacco 

Immunobiology Graduate Program, Iowa State University and USDA, ARS, National 

Animal Disease Center, Ames, Iowa 

Pasteurella multocida toxin (PMT) induces pre-osteoclast proliferation and osteoclast 

differentiation that leads to bone destruction. Developing B-cells and osteoclasts share 

the same microenvironment; therefore, it is hypothesized that the toxin may elicit 

alterations in B-cell progenitors in the bone marrow. To test this, mice were exposed to 

the toxin for 24 hours; peripheral blood, spleen, and bone marrow cells isolated, counted, 

and developing B-cell populations analyzed by flow cytometry. Toxin exposure resulted 

in a 5% increase in the percentage of B220+

CD19+IgM- cells in the bone marrow, as well 

as a 5% increase in the percentage of B220+

CD19+IgM+ cells in the peripheral blood. 

Conversely, percentages of B-cell subsets in the spleen were similar in toxin-treated and 

control mice. The increase in bone marrow B220+CD19+IgM- cells was not due to 

proliferation as determined by in vivo administration of bromodeoxyuridine (BrdU) 

during PMT exposure. Furthermore, PMT did not alter the expression of the antiapoptotic Bcl-2 as measured by intracellular staining. As B220+

 cells have been shown 

to serve as osteoclast precursors, we are currently investigating whether PMT induces 

osteoclastogenesis of bone marrow B220+

CD19+IgM- cells. 

Functional genomics analysis of infection by bovine viral diarrhea virus Type 

2 in a bovine B-cell lymphosarcoma cell line

John D. Neill and Julia F. Ridpath 

USDA, ARS, National Animal Disease Center, Ames, IA 50010 

Bovine viral diarrhea virus (BVDV), a major viral pathogen of cattle in the US, causes 

disease characterized by immunosuppression and lymphocytopenia. In addition, infection 

of a fetus in the first 150 days of pregnancy can give rise to a persistently infected calf 

that sheds the virus for life. This is a major means of dissemination of the virus to naïve 

animals and herds. BVDV is lymphotrophic and acute infection results in depletion of 

lymphoid tissues, even with BVDV strains of low virulence. The limited in vitro studies 

of host cell response done to date were performed in epithelial cells and compared 

cytopathic and noncytopathic strains without regard to virulence in vivo. Our research 

has shown that cytopathology in vitro does not correlate with virulence in vivo. We are 

taking a functional genomics approach, using serial analysis of gene expression (SAGE), 

to determine the molecular events that take place in the infected lymphoid cells that 

contribute to the disease syndromes observed in vivo. SAGE is a powerful technology 

that allows quantitation of transcripts in cells and direct comparisons between libraries 

and experiments. For these studies, we used the B-cell lymphosarcoma cell line BL-3 and 

compared virus strains of differing virulence (virulence was based on previous in vivo 

studies). To date, we have constructed and sequenced SAGE libraries from mRNA 

derived from non-infected cells, as well as cells infected with high and low virulence 

BVDV strains. Data analysis is ongoing. Preliminary results from the non-infected cells 

showed the normal metabolism of these cells with corresponding gene expression levels. 

When compared to results from BVDV-infected cells, a remodeling of cellular functions 

2003 ARS Immunology Research Worskshop 59 
was observed that reflected changes that, overall, benefited the replication of the virus. 

These effects included changes in protein translation, transport and ER processing, 

intracellular signaling, and some important housekeeping functions. Also observed was a 

decline in expression of proteins that are involved in detoxification, particularly reactive 

oxygen species. Some of these changes have suggested new lines of research that should 

lead to elucidation of the mechanism(s) behind immunosuppression and viral persistence. 

Immune responses controlling Toxoplasma gondii infection in pigs.

S.M. Nishi1, H. Dawson2, J.P. Dubey3, J.F. Urban2 and J.K. Lunney1. 1

USDA, ARS, Immunology and Disease Resistance Lab; and 2Nutrient Requirements and 

Functions Lab, BHNRC, and 3

Parasite Biology, Epidemiology and Systematics Lab, 

ANRI, Beltsville, MD 20705, USA. 

Toxoplasma gondii (Tg) infection induces a strong interferon-gamma (IFNg) dominated, 

T helper-1 (Th1) response in pigs with immune pathology that quickly resolvesPigs were 

infected with 4.5 x 106 Tg oocysts and various tissues were collected at 2, 4, 7 and 14 

days after infection (DAI). Tg organisms were detected in mesenteric lymph nodes 

(MLN) at 2DAI and, at 4 DAI, in liver, spleen, lung and ileum, by IHC and Tg specific 

Real-Time PCR. At the same infection times changes in inflammatory and Th1 immune 

mRNA gene expression were observed by Real-Time RT-PCR. Increased IFNg mRNA 

and protein expression were observed in MLN and hepato-splenic(HS) LN cells. Changes 

were observed in IL1b, IL-6, NRAMP1, IFNg, INDO and STAT-1 expression at 2DAI. 

Responses were more intense at 4DAI and also included TNFa, Arginase, IRF-1, IL-10, 

SOCS-1, SOCS-3, and IFNa activation. Liver showed the most intense changes in gene 

expression in response to Tg infection; PMBC had the lowest changes. High serum

haptoglobin and NO levels at 4DAI (p<0.01) confirms inflammatory and macrophage 

activation. Up-regulation of inflammatory and Th1-dominated markers indicates 

mechanisms involved in the host defense against acute toxoplasmosis. 

Supported by ARS funds, and National Pork Board and BRDC grants. 

Acute phase proteins in relation to disease and immunity: an avian paradigm

Narayan C. Rath 

USDA, ARS, PPPSRU, Fayetteville, AR 72701 

Acute phase proteins (APPs) are serum proteins elevated in response to a variety of 

physiological injuries including infection and inflammation. These are pathogen 

nonspecific proteins predominantly synthesized in the liver and other tissues such as 

endothelium, and modulate both innate and acquired immune responses. However, the 

physiological significance of different APPs is not well understood. In monitoring 

veterinary health the APPs are used as diagnostic and prognostic indicators. But their use 

to improve immunity has not been explored. The avian APP response and their 

significance are much less understood compared to mammals. The object of our study 

was to understand avian APP response and their possible significance. We induced 

experimental inflammation in young broiler chickens, by the use of intramuscular 

injection either of bacterial lipopolysacharides (LPS), gram positive bacterial cell walls, 

or croton oil, and compared the serum proteins profiles with sham injected control sera

2003 ARS Immunology Research Worskshop 60 
using polyacrylamide gel electrophoresis. Our results showed quantitative changes in 

several protein bands of which a band corresponding to MW of 65-66 kD was 

significantly elevated in response to inflammation (1-4). Further analyses using twodimensional gel electrophoresis and electroblotting onto PVDF membrane, the spots 

equivalent to 65 kD MW was N-terminal sequenced to yield a molecular homology to 

ovotransferrin (OVT) (3,4). Using an anti-serum transferrin antibody and western blot, 

the results further proved that the serum protein was the same as the egg white 

transferrin. We developed an enzyme-immunoassay to measure OVT concentrations and 

showed that the serum transferrin levels are elevated in response to a variety of bacterial 

and viral infections (5). To understand the possible significance of elevated OVT in 

relation to immunity, we studied its in vitro effects on macrophages and heterophil 

functions under non-stimulated and endotoxin stimulated conditions. Our results show

that OVT augmented the functional activity of both of these cell types as measured using 

nitric oxide, interleukin-6, matrix metalloproteinase, reactive oxygen production as 

indicators (6). Additionally, OVT also stimulated heterophil degranulation. These 

studies provide a paradigm of the possible usefulness of monitoring serum OVT to 

identify the presence of avian health problems and its function as an immunity modifying 

protein in poultry. 

References: 

Xie, H, Rath NC, Huff GR, Huff WE, Balog, JM, 2000. Effects of Salmonella 

typhimurium polysaccharide on broiler chicken. Poultry Sci 79:33-40. 

Xie H, Rath NC, Huff GR, Balog JM, Huff WE, 2001. Inflammation-induced changes in 

serum modulate chicken macrophage function. Vet Immunol Immunopathol. 80:225-235. 

Xie H, Rath NC, Newberry, L., Huff GR, Huff WE, Balog JM. 2001. Ovotransferrin, a 

major acute phase protein induced by inflammation. In: Current Progress in Avian 

Immunol. K.A. Schat ed. 302-307, American Assoc. Avian Pathologist, PA. 

Xie H, Huff GR, Huff WE, Balog JM, Holt P, Rath NC. 2002. Identification of

ovotransferrin as an acute phase protein in chickens. Poultry Sci. 81:112-120. 

Xie H, Newberry L, Clark FD, Huff WE, Huff GR, Balog JM, Rath NC, 2002. 

changes in serum ovotransferrin levels in chickens with experimentally induced 

inflammation and diseases. Avian Dis.46:122-131.

Xie H, Huff GR, Huff WE, Balog JM, Rath NC, 2002. Effects of ovotransferrin on 

chicken macrophages and heterophil-granulocytes. Dev Comp Immunol. 26:805-815. 

 

In vitro studies of BVDV strains in lymphoid cells lines reveal a third biotype

Julia F. Ridpath and John D. Neill 

USDA, ARS, National Animal Disease Center, Ames, IA 50010 

While bovine viral diarrhea viruses (BVDV) are highly lymphotrophic in vivo, in vitro 

they are studied in epithelial cells. BVDV are classified as cytopathic (cp) or 

noncytopathic (ncp) based on their activity in cultured epithelial cells. This does not 

2003 ARS Immunology Research Worskshop 61 
correlate with virulence in acute infections, as the most highly virulent BVDV are ncp. 

The purpose of this study was to determine if virulence in vivo could be correlated with 

growth characteristics in cultured lymphoid cells. A BL-3 cell line (derived from a B cell 

lymphosarcoma) free of BVDV and bovine leukemia virus was used. Cultures were 

infected with the highly virulent (hv) ncp BVDV2 strain 1373, the low virulent (lv) ncp 

BVDV2 strain 28508-5 or the lv cp BVDV2 strain 296c. Cell growth, proliferative state, 

ratio of live to dead cells and virus replication were monitored. Cell morphology was 

observed by light and electron microscopy. No significant differences were seen between 

noninfected cells and 28508-5 infected cells. Cytopathic effect (cpe), as defined by cell 

death, was observed in cells infected with 1373 and 296c. However, the cpe observed 

with the two strains were different. The cpe associated with 296c infection was apparent 

within the first 48 hours after infection and was associated with the condensation and 

fragmentation of most nuclei. Cpe associated with 1373 was not apparent until 96 hours 

after infection. Cells were non-proliferative but nuclei appeared morphologically normal. 

The cell cytoplasm became less dense and disorganization of mitochondria was observed. 

Nuclear condensation and fragmentation became common only in cells in the late stages 

of cpe. These results suggests that different mechanisms are involved in the cell death 

associated with lv cp strains and hv ncp virulent strains. Based on these findings BVDV 

may be grouped into three biotypes, cp (based on cpe in epithelial cells and cultured 

lymphoid cells within the first 48 hours after infection), ncp (based on lack of cpe in both 

epithelial and lymphoid cells lines and lymphocidal (based on cell death in lymphoid cell 

lines 96 hrs post infection). 

Local infection of VSV: pathogenesis, role of inflammatory cells and 

cytokines

Charles F.C. Scherer 1,2, William T. Golde 1, Douglas Gregg 1, D. Mark Estes2 and Luis 

L. Rodriguez1

 1

 USDA, ARS, Plum Island Animal Disease Center, Greenport, NY ; 2 College of 

Veterinary Medicine, University of Missouri, Columbia, MO 

Vesicular Stomatitis Virus (VSV) is negative-sense RNA virus that causes vesicular 

disease in cattle, horses and pigs. Little is known about the pathogenesis of this virus in 

livestock species. In order to better understand the pathogenesis, virus distribution and 

local immune response to VSV infection, 3 bovines were inoculated with the New Jersey 

strain of vesicular stomatitis virus (VSV-NJ), utilizing a coronary-band scarification 

procedure. Punch biopsies were obtained at 20 minutes, 6, 12, 24, and 48 hours postinoculation (hpi). Plasma and esophageal-pharyngeal fluid (OPF) were obtained prior to 

inoculation and at 24, 48 and 72 hpi. At 72 hours animals were euthanized and various 

tissues were collected including all major internal organs and various lymphoid tissues. 

Samples were assayed for virus isolation, presence of viral RNA by real-time RT-PCR 

and viral antigen identification by immunohistochemistry (IHC). Levels of mRNA for 

twelve cytokines were measured by real-time RT-PCR in the coronary band. 

Virus inoculation resulted in fever and vesicular lesions in the coronary bands by 48 hpi 

in all animals, with virus titers increasing over time from 1 log at 20 minutes to 6 logs 

TCID50 by 72 hpi. Virus was isolated only from lymph nodes draining the coronary 

bands (prescapular and popliteal). Virus was also isolated from other lymphoid tissues: 

2003 ARS Immunology Research Worskshop 62 
tonsil, retropharingeal, and parotid lymph nodes in one animal and tonsil and axilar 

lymph node in a second. Virus was never found in blood, OPF samples or in any other 

lymph node or internal organ tested. Viral antigens were detected predominantly in the 

stratum spinosum of the coronary band, in a few cells of the basal layer, and in the 

afferent and efferent sections of the local lymph nodes, with minimal involvement of 

germinal centers. Target cells in lymph nodes were morphologically compatible with 

monocytes. Local cytokine responses were dominated by an IFN alpha, beta, gamma and 

IL6, all showing increases of 3 to 4 fold by 6 hpi and 15-20 fold by 72 hpi. Levels of 

IL1β, IL2, IL4, IL10, IL12p40, IL13, IL15 and TNF alpha maintained basal levels up to 

24 hpi but increased 3-25 fold in all animals thereafter. Results demonstrate that infection 

with VSV-NJ was restricted to the local infected tissue and draining lymphoid tissue. A 

viral growth curve was observed in coronary band biopsies with increasing titers 12 to

72 hpi. These results provide an important contribution for understanding VSV 

pathogenesis and transmission in cattle. 

Characterizing mematode-induced Th2-dependent changes in intestinal function.

Terez Shea-Donohue, Aiping Zhao, Kathleen Madden, Motoko Morimoto, William

Gause, Dante Zarlenga and Joseph Urban. 

USDA, ARS, Nutrient Requirements & Functions Laboratory, Beltsville Human 

Nutrition Research Center, Beltsville, MD and US Department of Defense, Uniformed 

Service University of the Health Services, Bethesda, MD. 

The Th2 cytokines IL-4 and IL-13 are induced by parasitic worm infection and contribute 

to their expulsion through IL-4 receptor (R)-induced Stat6 activation. IL-4 binds the type 

1 and type 2 IL-4R that contain the IL-4R" chain and the common ( chain or the IL13Rα1 chain, respectively; IL-13 binds to the type 2 IL-4R and to the IL-13Rα2 chain 

that is not linked to Stat6. 

Aim: To determine the effects of intestinal nematodes on intestinal function. 

Methods Stripped mucosae were mounted in Ussing chambers to measure tissue 

permeability and concentration-dependent changes in short circuit current in response to 

glucose or various secretagogues, and strips of smooth muscle were suspended in organ 

baths and subjected to stimulation of enteric nerves. Expression of IL-4Rα, IL-13Rα1, 

and IL-13Rα2 mRNA was analyzed by real time PCR in specific mucosal cells collected 

by laser capture microdissection. 

Results: Infection of mice with 3 different nematodes (Heligmosomoides polygyrus, 

Nippostrongylus brasiliensis and Trichinella spiralis) elicits a stereotypic, Stat6-

dependent inhibition of glucose absorption and increased mucosal permeability leading to 

an elevation of fluid in the small intestine; these effects are mimicked by treatment with 

IL-4 and IL-13 and are Stat6-dependent. Mast cells play a key role in the pro secretory 

effects of IL-4 that are not shared by IL-13. Nematodes uniformly induces hyper 

contractility of intestinal smooth muscle by mechanisms that involve both Stat6-

dependent and Stat6-independent pathways; IL-13 has more potent effects on 

contractility than IL-4. Expression of IL-4Rα, IL-13Rα1 and IL-13Rα2 mRNA was 

2003 ARS Immunology Research Worskshop 63 
evident in epithelial and smooth muscle cells. Infection of pigs with the nematode 

Ascaris suum revealed similar Th2-driven changes in intestinal function associated with 

expulsion of 4th stage larvae. 

Conclusion: Nematode infection of the small intestine elicits a characteristic "weep and 

sweep" response that facilitates worm clearance. IL-4 and IL-13 play a critical role in the 

host response to nematode infection by altering gut function through receptor-mediated 

activation of Stat6 pathways. Currently, cloned soluble IL-4Rα, IL-13Rα1, and IL13Rα2 are being used in swine as molecular scavengers to 1) delineate the role of IL-4 

and IL-13 in worm clearance and intestinal function; 2) as antagonists and/or agonists of 

the classic Th2 response and; 3) to mediate IL-13 induced as a model of human allergic 

disease. 

Modulatory effects of IFN-γ, IL-10 and TGF-β in cattle infected with 

Mycobacterium paratuberculosis

J. R. Stabel and M. S. Khalifeh 

USDA, ARS, National Animal Disease Center, Ames, IA 50010 

Johne’s disease is an important disease that results in great economic losses for both dairy 

and beef production in the United States and worldwide. The disease progresses through 

distinct stages, a subclinical stage where there are no clinical signs and a clinical stage 

that is characterized by progressive symptoms associated with chronic shedding of high 

levels of bacteria in the feces along with severe diarrhea and concomitant weight loss. 

The host immune response to M. paratuberculosis is paradoxical, with strong cellmediated immune responses during subclinical stages and strong humoral responses 

during clinical stages of the disease. It is possible that immune modulation of an effective 

cell-mediated immune response may play an important role in disease progression. We 

hypothesized that the clinical stage of Johne’s disease is mediated by production of 

cytokines such as TGF-β and IL-10 that downregulate IFN-γ production and interfere 

with an effective cell-mediated immune response. Therefore, ileum, ileal cecal junction, 

ilealcecal lymph node and mesenteric lymph node tissues from the healthy, subclinical or 

clinical animals were collected and analyzed for the presence of TGF-β, IL-10 and IFN-γ

mRNA by quantitative RT-cPCR. The results show that TGF-β and IL-10 mRNA levels 

in animals that have progressed to the clinical stage of the disease is higher than that 

found in subclincal and healthy animals, whereas IFN-γ is higher in subclinical animals. 

Interferon-γ plays a significant role in the control of mycobacterial infections, including 

Mycobacterium paratuberculosis. However, the contribution of other immunoregulatory 

cytokines such as IL-10 and TGF-β in Johne’s disease has not been investigated as yet. In 

the present study, we examined the effects of in vivo and in vitro infection with M. 

paratuberculosis on the production of IFN-γ, IL-10 and TGF-β by peripheral blood 

mononuclear cells (PBMC). We also examined the effects of exogenous IFN-γ, IL-10 and 

TGF-β on M. paratuberculosis survival in the cell cultures. PBMC obtained from

naturally infected cows, regardless of their disease status, specifically upregulated IL-10 

and TGF-β in culture supernatants in response to stimulation with live M. 

paratuberculosis. Non-stimulated PBMC recovered from subclinically infected animals 

secreted the lowest levels of TGF- but after stimulation with live M. paratuberculosis

2003 ARS Immunology Research Worskshop 64 
TGF-β levels in the culture supernatants increased to levels similar to that produced by 

PBMC from healthy animals. Compared to healthy cows, naturally infected animals had 

higher numbers of viable M. paratuberculosis recovered from their cultures after in vitro 

infection with M. paratuberculosis. The addition of exogenous IL-10 and TGF-β to 

PBMC isolated from healthy cows inhibited the bactericidal activity of these cells as 

evidenced by the increased number of viable M. paratuberculosis recovered from those 

cultures compared to cell cultures containing medium alone. These data suggest an 

important immune regulatory role of IL-10 and TGF-β during infection with M. 

paratuberculosis that may be directly related to their effects on macrophage activation 

and killing of M. paratuberculosis. 

Bovine T cell response to antigens of Neospora caninum.

Wenbin Tuo1

, Ray Fetterer2, Pat Boyd1, Lou Gasbarre3, JP Dubey2. 1

 USDA, ARS, Immunology and Disease Resistance Lab, 2Animal Parasitic Diseases Lab, 

3

Bovine Functional Genomics Lab, ANRI, BARC-East, Beltsville, MD 

See abstract on page 24.

Ascaris Infection as a Marker of Local Mucosal Immunity in Pigs.

Joseph Urban, Ethiopia Beshah, Motoko Morimoto, Gloria Solano-Aguilar, Norberta 

Schoene, Dante Zarlenga, Ben Mathews, Terez Shea-Donohue, and Harry Dawson. 

USDA, ARS, Beltsville Human Nutrition Research Center and Plant Science Institute, 

Beltsville, MD. 

Ascaris spp. are ubiquitous in swine with significant economic consequences to the industry, 

and are the most prevalent helminth infection in man worldwide. The migration of this 

nematode parasite from the cecum to the liver, lung and small intestine along with its ability to 

induce immunity and inflammation provides a tool for analysis of local responses at several 

mucosal surfaces. The Th2-like response to A. suum in pigs that is characterized by mucosal 

mast cell and goblet cell hyperplasia, eosinophilia and reaginic antibody production was 

supported by the demonstration of a pattern of cytokine gene expression consistent with a 

polarized Th2-related immune response in the lymph nodes draining the sites of infection 

using a series of 24 gene probes for Th1 and Th2-associated products. Alveolar macrophages 

obtained from the lungs of A. suum-infected pigs had reduced capacity to phagocytise killed 

Staphylococcus aureus but did release enhanced levels of super oxide dismutase indicating a 

functional skewing of immunity in the lung. Likewise, expulsion of the migrating 4th stage 

larvae from the jejunum of infected pigs was associated with a “weep and sweep” 

phenomenon in the small intestine characterized by increased fluid accumulation in the lumen 

and enhanced smooth muscle contractility. Micro array analysis of parasites that survived 

expulsion revealed a more robust expression of transcripts for glyceraldehydes 3 phosphate 

dehydrogenase, succinate dehydrogenase and muscle proteins. Application of sensitive new 

technologies for evaluation of gene expression of the host and parasite have made the pig a

more accurate predictor of immunity in this species and as an attractive model of immune 

function in humans. 

2003 ARS Immunology Research Worskshop 65 
Experimental Mycobacterium avium subsp. paratuberculosis infection of 

calves: early induction of a concurrent humoral and cellular immune 

response.

W. R. Waters

1*, J. M. Miller1

, M. V. Palmer1, J. R. Stabel1, D. E. Jones2, K. A. 

Koistinen2

, E. M. Steadham2, M. J. Hamilton3, W. C. Davis3, and J. P. Bannantine1

1

USDA, ARS, National Animal Disease Center, Ames, IA; 2Iowa State University, 

Ames, IA, 3

Washington State University, Pullman, WA

Johne’s disease (paratuberculosis) of cattle is widespread and causes significant 

economic losses for producers due to decreased production and poor health of affected 

animals. The chronic nature of the disease and the lack of a reproducible model of 

infection hinder research efforts. In the present study, instillation of Mycobacterium 

avium subsp. paratuberculosis into the tonsillar crypts of neonatal calves resulted in 

chronic infection with peripheral colonization detected by ante-mortem culture of feces 

and post-mortem (320 d postchallenge) culture of intestinal tissues. Antigen-specific 

blastogenic, IFN-γ, and nitric oxide responses by blood mononuclear cells from infected 

calves exceeded prechallenge responses beginning 194 d postchallenge. Upon in vitro 

stimulation with paratuberculosis antigens, CD4+

 cells from infected calves proliferated, 

produced IFN-γ, and increased expression of CD26 and CD45RO (i.e., indicative of an 

activated memory phenotype). Utilizing a lipoarabinomannan-based ELISA, specific 

serum immunoglobulin was detected as early as 134 d postchallenge and generally 

increased after this time point. Two antigens of ~50-kDa and ~60-kDa were particularly 

immunodominant early in infection as shown by immunoblot with serum collected within 

2 weeks postchallenge. Findings indicate that the intratonsillar inoculation route will 

prove useful as an experimental model for paratuberculosis pathogenesis, diagnosis, and 

vaccine efficacy studies. Additionally, this study confirms that mycobacteria-specific 

antibody is detectable early in the course of experimental Johne’s disease, even 

preceeding the development of specific cell-mediated responses. 

Mechanism for age-associated upregulation in COX-2 expression: role of 

NFkB.

Wu D, Marko M, Claycombe KJ, Eric Paulson and Meydani SN. 

USDA, ARS, Nutritional Immunology Laboratory, JMHNRCA at Tufts University, 

Boston, MA 02111 

Increased prostaglandin E2 (PGE2) production has been shown to contribute to ageassociated decline in T cell function as well as dysregulated inflammatory response. We 

previously demonstrated that macrophages (Mφ) from old mice have significantly higher 

cyclooxygenase (COX) activity and COX-2 expression compared to young mice. We

further showed that the higher COX-2 expression was due to transcriptional upregulation 

rather than a change in COX-2 degradation with aging. The purpose of the present study 

was to determine the mechanism of age-related upregulation in COX-2. We hypothesized 

that the activation of transcription factors may undergo a change with aging, leading to 

altered COX-2 transcription. The results demonstrate that LPS-initiated activation of 

2003 ARS Immunology Research Worskshop 66 
NFκB, but not AP-1 and CREB, is higher in old than in young mice. No significant age 

difference was observed in the activity of MAPK P38, JNK, or ERK. Inhibition of NFκB 

activation decreased PGE2 production, COX activity, COX-2 protein and mRNA 

expression. These reductions were more dramatic in old Mφ compared to that of young 

Mφ. We further showed that old Mφ have higher IκB degradation in cytoplasm and p65 

translocation to nucleus compared to young Mφ. Thus the age-associated up regulation of 

COX-2 transcription in Mφ is mediated through higher NFκB activation, which in turn is 

due to increase in IκB degradation. Supported by NIA grant # RO1AG09140-09 

2003 ARS Immunology Research Worskshop 67 
December 3, 2003

Immunity and Nutrition 

Real-time PCR arrays can delineate immunological and nutrition-related 

gene expression in swine.

Harry Dawson 

USDA, ARS, Nutrient Requirements & Functions Laboratory, Beltsville Human 

Nutrition Research Center, Beltsville, MD. 

See abstract on page 34.

Dietary modulation of host mucosal immunity against avian coccidiosis. 

Dalloul1

, R. A., Lillehoj2, H. S. and John A. Doerr1. 1

Department of Animal and Avian Sciences, University of Maryland, College Park, MD. 

20742. 2

 USDA, ARS, Animal Parasitic Diseases Laboratory, ANRI, Beltsville, MD. 

20705.

Infectious diseases constitute one of the greatest threats to the viability of the food animal 

industry. Commercial broilers, in particular, have a higher risk of acquiring contagious 

diseases than other livestock due to intensive collective farming practices. For example, 

avian coccidiosis is a major parasitic disease of significant economic importance and the 

U.S. industry spends greater than $800 million annually in medication. In the absence of 

efficient vaccines to control this disease and the emergence of new antigenic variants of 

Eimeria, the broiler industry has relied upon prophylactic medication. However, anticoccidial drugs are expensive and their effectiveness is hindered by widespread parasite 

drug resistance and the high cost of new drug development. Moreover, recent banning of 

growth promoters and anti-coccidial drugs in commercial animal production in Europe 

and parts of US will eventually force the industry to eliminate this practice and will 

require the development of non-chemical methods for coccidiosis control. Thus, 

probiotics represent an important alternative for antibiotic use in poultry production. 

However, the exact modes of action in which probiotics achieve such protective roles 

need to be better elucidated to improve effectiveness. Previously we have shown that 

resistance in broilers to Eimeria acervulina (EA) was significantly reduced by vitamin A 

deficiency and enhanced by a probiotic (Lactobacillus-based). In the present 2 x 2 

factorial study, a broiler starter ration was amended for vitamin A (control, C, or 

deficient, A) and probiotic status (-, P) to investigate their modulatory effects on the host 

immune system. Birds were inoculated orally with EA oocysts and disease susceptibility 

was evaluated by assessing fecal oocyst shedding. Humoral and local cellular mediated 

immunity were assessed by evaluating antibody and cytokine (IFN-γ and IL-2) levels in 

sera and intestinal secretions on a 3-day interval following inoculation. Fecal oocyst 

shedding was highest (P < 0.05) in A- birds, followed by AP, C- and CP. Feeding the 

probiotic reduced shed oocysts by 20% in A fed birds and by 26% in C fed birds. 

Intestinal IFN-γ was relatively constant in all treatment groups except for A- where it 

declined steadily and was lower (P < 0.05) from day 6 on. Serum IFN-γ levels fluctuated 

within each treatment and over time were not revealing. Intestinal IL-2 was highest in CP 

2003 ARS Immunology Research Worskshop 68 
birds at 3 and 9 dpi, and lowest in A- birds at 3, 9, and 12 dpi (P < 0.05); no difference 

between treatments was found at 6 dpi (P > 0.05). Eimeria-specific intestinal Ab level 

was constant (P > 0.05) in C- birds, but increased with time (P < 0.05) in A-, AP and CP 

birds. Serum Ab levels were also constant in A- and CP, but increased (P < 0.05) in Cand AP after 6 dpi. The data demonstrate for the first time a probiotic-enhanced 

immunity in vitamin A-deficient birds. It is also the first study to demonstrate the 

probiotic effect on local cell-mediated immunity of chickens, best manifested by apparent 

lower intestinal invasion and development by EA, based on higher IL-2 secretion and 

lower EA oocyst production. The new information concerning the immunological 

mechanism involved in the probiotic-induced immune enhancement would promote the 

development of novel ways to modulate the immune response to enhance the health and 

productivity of poultry. 

The effect of vitamin E on secondary bacterial infection following influenza 

infection in young and old mice

Raina Gay1, Sung Nim Han1, Melissa Marko1, Sarah Belisle1, Roderick Bronson2, Simin 

N Meydani1

. 1

 USDA, ARS, JMHNRCA at Tufts University, Boston, MA, 2 Tufts University School of 

Veterinary Medicine, North Grafton, MA 

In the U.S. alone, influenza accounts for over 10,000 deaths annually and over 40,000 

deaths during epidemic years. For example, individuals 65 years and older accounted for 

89% of all influenza associated deaths in 1992. These deaths were not typically due to 

influenza infection alone, but from the development of complications due to influenza 

infection. Staphylococcus aureus is commonly associated with influenza infection and 

can cause sudden and serious illness, especially in the elderly. Previously, we showed 

that vitamin E supplementation significantly improves the antioxidant status and immune 

response in aged mice and humans. We further showed that vitamin E supplementation 

reduces influenza viral infection and improves antioxidant status in aged mice. However, 

little is known about the potentially deadly bacterial infections that commonly follow 

influenza infection in the aged. The goal of this study was to determine the efficacy of 

vitamin E supplementation on secondary bacterial infection following influenza infection 

in young and old mice. This was tested by feeding young and old C57BL/6 mice semisynthetic diets containing adequate (30 ppm) or high (500 ppm) levels of vitamin E and 

determining viral and bacterial titers after primary influenza A or S. aureus infection, or 

following a secondary S. aureus infection subsequent to an influenza infection. In this 

animal model, age did not have a significant effect on S. aureus infection alone or S. 

aureus infection following influenza infection. Supplementation with vitamin E did not 

have a significant effect on S. aureus infection alone. Priming with influenza infection 

significantly increased S. aureus colony counts in the lungs of infected mice fed the 

control diet. Vitamin E supplementation successfully abolished the priming effect of 

influenza on S. aureus infection. Future studies will determine the mechanism of vitamin 

E’s protective effect. 

Supported by a 2002 Glenn/AFAR Scholarship for Research in the Biology of Aging by 

the American Federation for Aging Research and USDA contract 58-1950-9-001. 

2003 ARS Immunology Research Worskshop 69 
Changes in gene expression profiles of T cells by age and vitamin E.

Sung Nim Han1

, Oskar Adolfsson1, Cheol-Koo Lee2, Tomas A Prolla2, Jose Ordovas1, 

Simin N Meydani1

. 1

 USDA, ARS, JMHNRCA at Tufts University, Boston, MA, 2 University of Wisconsin, 

Madison, WI 

Aging is associated with dysregulation of immune cells particularly that of T cells. 

Previous studies indicated that vitamin E improves T cell function in part by directly 

affecting T cells. We studied the gene expression profile of T cells to better understand 

the underlying mechanisms of age and E-induced changes in T cell functions. Young and 

old C57BL mice were fed diets containing 30 ppm (control) or 500 ppm (E) of vitamin E 

for 4 weeks. T cells were purified from splenocytes by negative selection using magnetic 

beads (anti-Mac-1 and anti-MHC class II), then cultured with media (unstimulated) or

plate-bound anti-CD3 and soluble anti-CD28 (stimulated) for 2 hours. Gene expression 

profile was assessed using microarray analysis. Genes showing more than 2 fold changes, 

p < 0.05, and with at least 1 present call were selected. In unstimulated T cells, 43 genes 

were expressed significantly higher in old, mainly immunoglobulin genes, while 22 genes 

were expressed significantly higher in young including T cell receptor related genes. 

Upon stimulation, 103 genes were upregulated in at least one age or diet group (16 genes 

in all age and diet groups). Response to stimulation was significantly affected by age (40 

genes) and vitamin E (24 genes). Significantly lower expression of T receptor alpha chain 

and factors involved in the signal transduction pathways were observed with aging. 

Significantly higher expression of suppressor of cytokine signaling (SOCS) 3 and lower 

expression of growth factor independent (gfi) 1 were observed with aging. 

Overexpression of SOCS3 has been shown to suppress T cell proliferation and IL-2 

production and gfi1 has been reported to down regulate SOCS3. Expressions of IL-2 

increased significantly in both young and old T cells while that of IL-4 decreased in old T 

cells by vitamin E. These findings suggest that: 1) aging has significant effects on signal 

transduction pathways in T cells, and 2) vitamin E has significant effect on Th1/2 

balance. 

Supported by NIA grant #RO1AG09140-09, USDA contract 58-1950-9-001, and gifts 

from Roche, Inc. 

Stress-related chronic disease in turkeys and the effects of nutritional and 

environmental immunomodulation. 

Gerry Huff, Bill Huff, Narayan Rath, and Janice Balog. 

USDA, ARS, Poultry Production and Products Safety Research, Fayetteville, AR 72701. 

Our research using the synthetic glucocorticoid, dexamethasone (DEX), to suppress the 

immune response of turkeys to low doses of opportunistic bacteria has led to the 

development of a number of nutritional and environmental strategies for decreasing the 

impact of stress-induced immunosupression. The immunosuppressive effects of stress 

appear to be greater in male turkeys than in females. We have found that female turkeys 

are more resistant to colibacillosis and respiratory disease in a DEX-E.coli challenge 

infection than are males. The anti-bacterial activity of monocytes from male and female

turkeys treated with 0.5 or 2.0 mg/Kg DEX was measured by monitoring the percentage 

2003 ARS Immunology Research Worskshop 70 
of infected cells over time. Antibacterial activity was significantly decreased at both 

concentrations of DEX at both 8 and 16 hours post-infection in both sexes combined and 

was significantly lower in males as compared to females, suggesting that stress can 

reduce the ability of turkey mononuclear cells to kill bacteria, and that the difference in 

this function may be relevant to the sex-related difference in disease resistance. Water 

supplementation with vitamin D3, vitamin E, sodium salicylate, and ß-1,3/1,6-glucan, a 

helical polysaccharide derived from the cell wall of Saccharomyces cervisiae, have all 

shown immunomodulatory effects on male turkeys challenged with E. coli in our model. 

We have shown that excess handling or environmental enrichment during the first 2 

weeks after hatch can lead to decreased resistance later in life. Our model has further 

suggested that a small population of male turkeys is highly susceptible to the 

immunosuppressive effects of stress, and that these individuals can be identified by their 

performance in a behavioral test utilizing a T-maze at 3 days of age. These individuals 

are also more susceptible to the immunosuppressive effects of excess environmental 

stimulation during the first two weeks after hatch. These studies illustrate the dramatic 

effects that stress can have on both production values and resistance to opportunistic 

infection, and suggest that a combination of genetic selection, nutritional 

immunomodulation, and environmental strategies to reduce stress may lead to healthier 

birds and a safer and a more wholesome product. 

Purified $-glucan as an abiotic feed additive up-regulates heterophil function 

in immature chickens.

V.K. Lowry1*, M.B. Farnell2, P.J. Ferro1, A. Bahl3, M.H. Kogut4

1

Texas A&M University, Departments of Anatomy and Public Health & Poultry Science, 

respectively, College Station, TX 77843, 2

Institutes for Animal Health, Compton, 

Berkshire RG20, 7NN, UK, 3

ImmuDyne, Inc., 11200 Wilcrest Green Dr., Houston, TX 

77042, 4

USDA, ARS,SPARC, 2881 F&F Road, College Station, TX 77845. 

Functionally, the innate immune system of immature chickens is inefficient during the 

first week post-hatch. Pathogens, such as Salmonella entriica Serovar Enteritidis (SE), 

are able to invade various tissues due to this immunological inefficiency of the immature 

chicken. The objective of the present study was to evaluate the effect of purified $-glucan 

as an immunomodulator of the innate immune response, specifically the heterophil. The 

functional efficiency of heterophils isolated from immature chickens fed a $-glucan 

ration was significantly (P<0.05) up-regulated when compared to heterophils isolated 

from chickens fed a control ration as determined with an array of functional assays. 

Phagocytosis, bactericidal killing and oxidative burst were significantly increased in 

heterophils isolated from chickens fed the purified $-glucan ration (P<0.05). To our 

knowledge this is the first report of a purified $-glucan feed additive significantly upregulating the functional abilities of heterophils isolated from immature chickens against 

an invading pathogen, SE. 

Altered structure, function and allergenicity of peanut proteins due to 

processing 

Soheila J. Maleki 

2003 ARS Immunology Research Worskshop 71 
USDA, ARS, Food Processing and Sensory Quality Research Unit, Southern Regional 

Research Center, New Orleans, LA 70124, Fax: 504-286-4419, 

sjmaleki@srrc.ars.usda.gov 

There has been a dramatic increase in the prevalence of IgE mediated allergy over the 

past decade, which is one of the reasons that it is viewed as an area of increasing 

importance. During this time a significant, growing, and costly, portion of the FDA's food 

recalls have been due to mislabeled products that contain food allergens. Meanwhile, 

little is known about the reason why certain foods contain allergens and others are 

considered hypoallergenic. Some classic characteristics associated with food allergens are 

resistance to enzymatic digestion and heat, stimulation of T cell proliferation, IgE binding 

and abundance in food. Certain enzymatic functions have also been associate with 

allergenicity of proteins. However, not much is known about what happens to the 

allergenicity of food products after processing. In this study, roasted whole peanut 

proteins were shown to be less soluble, more resistant to digestive enzymes and bound 

higher levels of IgE than raw whole peanut proteins. Mice were more likely to be 

sensitized when treated with roasted peanuts than with raw peanuts and had higher 

vascular permeability when skin tested with roasted peanuts. The major peanut allergens 

Ara h 1 and Ara h 2 were then purified from both raw and roasted peanuts and examined 

at a molecular level for alterations that may explain the in vivo observations. Following 

roasting, Ara h 1 was shown to form irreversible trimers that are extremely resistant to 

digestion and bind higher levels of IgE than Ara h 1 from raw peanuts. The amino acid 

sequence of a second major peanut allergen, Ara h 2 was found to be homologous to the 

family of trypsin/alpha-amylase inhibitors. Most significantly, while the secondary 

structure remained unaltered, Ara h 2, purified from roasted peanuts was several fold 

more active as a trypsin inhibitor and bound higher levels of IgE than the Ara h 2 purified 

from raw peanuts. Our findings suggested that the structural and functional changes due 

to food processing contribute to increased allergenic properties of peanut proteins and 

may indeed enhance the possibility of becoming sensitized to peanuts. 

Interpretive summary

It is not known why certain foods are allergenic and others are not. Some classic 

characteristics associated with food allergens are resistance to digestion and heat, 

stimulation of T cell proliferation, IgE binding and abundance in a food. Certain 

enzymatic functions have also been associate with allergenicity of proteins. However, not 

much is known about what happens to the allergenicity of food products after 

processing. Here the consequences of roasting on the structural, functional, 

immunological and allergenic properties of peanut proteins were examined. Roasting 

was found to alter the structure and function of peanut allergens Ara h 1 and Ara h 2. In 

addition, whole peanut proteins from roasted peanuts were more resistant to degradation 

by digestive enzymes and more likely to immunize mice than raw peanut proteins. Our 

findings suggested that the structural and functional changes caused by food processing 

contribute to increased allergenic properties of peanut proteins and may indeed enhance 

the possibility of becoming sensitized to peanuts. 

2003 ARS Immunology Research Worskshop 72 
Aging, immune response, and infectious disease: molecular mechanisms and 

reversal by nutrient intervention.

Simin Nikbin Meydani, DVM, PhD, Dayong Wu, MD, PhD, Sung Nim Han, PhD, RD, 

Raina T. Gay. 

USDA, ARS, Nutritional Immunology Laboratory, Jean Mayer Human Nutrition 

Research Center on Aging at Tufts University, Boston , MA. 

See abstract on page 32.

Nutritional plane affects antigen-induced proliferation of lymphocyte subsets 

in milk replacer-fed calves vaccinated with M. bovis bacillus Calmette-Guerin 

(BCG). 

B. J. Nonnecke1, M. R. Foote2, M .A. Fowler3, B.L. Miller

3

, W. R. Waters4

1

USDA, ARS, Periparturient Diseases of Cattle Research Unit, NADC, Ames, IA, 2Iowa 

State University, Ames, IA, 3

Land O’ Lakes, Inc., Webster City, IA and 4 USDA, ARS, 

Bacterial Diseases of Livestock Research Unit, NADC, Ames, IA. 

See abstract on page 33.

Leukocyte migration and metabolism in adipose tissue of leukocyte adhesion 

receptor deficient mice.

Rebecca Robker, Robert Collins, Nancy Butte, Harry Mersmann and C. Wayne Smith

Section of Leukocyte Biology, Children’s Nutrition Research Center, Department of 

Pediatrics, Baylor College of Medicine, Houston, Texas 77030. 

ICAM-1 (CD54) and Mac-1 (CD11b) mediate the migration of leukocytes into tissues 

where they influence tissue function. Our studies sought to determine whether ICAM-1 

and Mac-1 have an intrinsic role within adipose tissue by 1) analyzing the expression of 

ICAM-1 in adipose tissue, 2) identifying leukocyte populations within adipose tissue, and 

3) determining whether ICAM-1 and Mac-1 mutant mice exhibit abnormal numbers of 

adipose tissue leukocytes or altered metabolism. In wildtype mice ICAM-1 was 

expressed in adipose tissue and localized to the vascular endothelium. ICAM-1 mRNA 

was detected in isolated adipocytes, but surface staining of the adipocyte was not evident, 

though cultured adipocyte supernate was positive for soluble ICAM-1. ICAM-1 and 

Mac-1 do not appear to be essential, however, for adipose tissue homeostasis since 

ICAM-1 and Mac-1 mutant mice exhibited no alterations in body fat deposition or

adipocyte metabolism. Macrophages were observed to be a prevalent cell type within the 

stromal-vascular cell fraction of adipose tissue, and adipose tissue and plasma from obese 

mice exhibited significant elevations in MCP-1. Macrophage numbers in adhesion 

molecule deficient mice did not differ from controls. However, gender-specific 

differences were observed, with adipose tissue from female mice containing significantly 

more macrophages than males. Our results are in contract to a previous report of 

increased obesity in ICAM-1 and Mac-1 mutant mice, but demonstrate leukocyte 

populations within adipose tissue which must be considered in light of increasing 

evidence of immune-modulation of body weight. 

The effect of human-derived probiotic bacteria on the immune and intestinal 

function of pigs

2003 ARS Immunology Research Worskshop 73 
Gloria Solano-Aguilar*1

, Harry Dawson1, Tonya Ledbetter1, Terez Shea-Donohue1, 

Norberta Schoene1

, Jeff Call

2

, Ethiopia Beshah3, William Hare3 and Joseph Urban1. 1

 USDA, ARS, Beltsville Human Nutrition Research Center, Beltsville MD; 2 USDA, 

ARS, Eastern Regional Research Center, Wyndmoor, PA 19038; 3USDA, ARS, Animal 

&Natural Resources Institute; Beltsville, MD 20705. 

See abstract on page 30.

Vitamin A deficiency decreases development of Th1 memory cells following 

adoptive transfer of TCR-transgenic T cells.

Charles B. Stephensen, Xiaowen Jiang, Tammy Freytag 

U.S.D.A. Western Human Nutrition Research Center at the University of California, 

Davis, and Nutrition Department, University of California, Davis, CA 95616 

Vitamin A deficiency enhances production of Th1 cytokines, particularly IFN-(, during 

the memory and effector phases of the immune response. This had led to the presumption 

that vitamin A deficiency biases T-helper cell development in the same direction. To test 

this hypothesis we adoptively transferred naïve T cells from DO11.10 TCR-transgenic 

mice to vitamin A-deficient or control BALB/c recipients. Recipients were then 

immunized with the cognate peptide antigen for the TCR-transgenic T cells (OVA323-

339) and the Th1/Th2 phenotype of the transferred cells characterized by intracellular 

cytokine staining following in vitro stimulation with phorbol myristate acetate and 

ionomycin. In contrast to expectations, the percentages of IFN-(-positive cells (9.0 "

0.5%; mean " standard error) and IL-2-positive cells (39.2 " 2.3%) in draining lymph 

nodes of deficient mice were both lower than in control mice (11.3 " 0.4%, p = 0.002 and 

46.4 " 2.1%, p = 0.028, respectively). In addition, the percentage of IL-10 positive cells 

was higher in vitamin A-deficient mice (3.71 " 0.24%) than it was in control mice (1.74 

" 0.22 %; p < 0.001). IFN-( and IL-2 concentrations were also lower in supernatants 

from splenocyte cultures restimulated with OVA323-339 peptide antigen. Thus vitamin 

A deficiency at the time of initial antigen exposure diminishes the development of Th1 

memory cells and enhances the development of IL-10-producing Th2 memory cells. 

2003 ARS Immunology Research Worskshop 74 
Immunity and Disease Resistance

Relationship of the probability of disease incidence with immunity

Gamal A. Abdel-Azim1*, Albert E. Freeman1

, Error! Bookmark not defined.M. E. 

Kehrli, Jr.2

, S. C. Kelm1, J. L. Burton3, A. L. Kuck1, and S. Schnell 

1

 Department of Animal Sciences, Iowa State University, Ames, IA; 2USDA, ARS, 

Metabolic Diseases and Immunology Research Unit, National Animal Disease Center, 

Ames, IA; 3

 Michigan State University, Department of Animal Science, Immunogenetics 

Laboratory and Center for Animal Functional GenomicsEast Lansing, MI 48824; *

Current address: Cooperative Resources International, Shawano, WI. 

Health data in Holstein cows sired by a selected group of sires were analyzed as 

categorical traits by a threshold model. Data were collected from 174 herds and included 

incidences on 18,000 daughters of 1365 sires. Breeding values and genetic parameters 

for disease incidence were estimated. Probability of disease incidence in daughters as 

values associated with their sires were computed. The immune competence of the sires

had been previously tested in a model of glucocorticoid immunosuppression. Statistical 

relationships between daughter health and immune response traits in sires were 

investigated. Heritability estimated by the threshold model was 0.277, an estimate 

sufficiently high to indicate the great potential of selection for disease resistance. 

Evidence of association between probability of disease incidence and seven immune 

response traits was found, which motivates and justifies the indirect selection of dairy 

sires for disease resistance. Indirect selection for disease resistance saves time and 

money, and could serve as a first guide for what bulls to select for health until young 

bulls have been progeny tested. Further research is warranted to develop indirect models 

of selection for disease resistance. 

Correlations between disease resistance, pathogen levels, and clearance rate 

in channel catfish

Lanie Bilodeau

1

, Brian Small1, David Wise

2

, and William Wolters

1

1

USDA-ARS, Catfish Genetics Research Unit, Stoneville, MS; 2Mississippi State 

University, MAFES, Stoneville, MS 

Enteric septicemia of catfish (ESC) is the most prevalent disease affecting commercial 

catfish farms. The USDA-ARS Catfish Genetic Research Unit has an ongoing program

for genetic improvement of disease resistance amongst other economically important 

traits. Susceptibility to the causative agent of ESC, the bacterium Edwardsiella ictaluri, 

appears to be consistent within each family/spawn of NWAC103 channel catfish. 

Real-time PCR technology was utilized to measure differences in bacterial loads and 

clearance rates for 6 ESC-susceptible and resistant families of channel catfish during 

immersion challenge. Fish from resistant families had increased survivorship during the

challenge when compared to susceptible families. Significant differences (p < 0.05) in the 

quantity of bacterial DNA between resistant and susceptible families were evident for 

both blood and spleen tissue 5 days following exposure to E. ictaluri. Mean quantities of 

2003 ARS Immunology Research Worskshop 75 
bacterial cell equivalents per 100 uL of blood at 5 days post-exposure were 2.84 x 105

"

143123 for the susceptible families and 496 " 455 for the resistant families. Significant

differences in spleen tissue also occurred on day 12, with fish from the resistant families 

having higher (p < 0.05) levels of bacteria than fish from the susceptible families. 

Overall, families that are susceptible to ESC carried higher levels of bacterial DNA in 

their blood than resistant families. 

Pathogen clearance differed between the two sets of families. Clearance was evident by 

day 12 in fish from the susceptible families and bacterial levels continued to decrease 

throughout the remainder of the trial. However, no significant clearance was evident in 

fish from the resistant families. This may be due to chronic low levels of infection that

did not trigger a clearance response from the immune system. The innate immune system

may suppress infection only in resistant families, preventing an acute host-response 

whereas in susceptible families, acute infection occurs and may secondarily be cleared.

Clearance rate does not appear to significantly affect mortality. 

Heterophils isolated from chickens resistant to extraintestinal Salmonella 

enteritidis infection express higher levels of pro-inflammatory cytokine 

mRNA following infection than heterophils from susceptible chickens

Pamela J. Ferro1, Christina L. Swaggerty2, Pete Kaiser3, Igal Y. Pevzner4 and Michael H. 

Kogut2

1

Texas A&M University, Poultry Science, College Station, TX 77843, USA 

2

USDA-ARS, SPARC, 2881 F&B Road, College Station, TX 77845, USA 

3

Institute for Animal Health, Compton, Berkshire RG20 7NN, UK 

4

Cobb-Vantress Inc., Siloam Springs, AR 72761-1030, USA 

See abstract on page 37. 

Genetic and immunologic basis for resistance to GI nematode infections

Louis C. Gasbarre, Tad Sonstegard, Ricardo Araujo, and Curt Van Tassell 

USDA, ARS, Bovine Functional Genomics Laboratory, ANRI, BARC, Beltsville MD 

See abstract on page 36.

2003 ARS Immunology Research Worskshop 76 
High-throughput molecular approaches to identify coccidiosis disease 

resistance genes and protective immunological mechanisms in avian 

coccidiosis. 

Lillehoj

1

, Hyun S., Min1, Wongi, Matthews2, Benjamin F., and Emara3, Marlene. 

1

 USDA, ARS, Animal Parasitic Diseases Laboratory, ANRI, and 2 USDA, ARS, 

Soybean Genomics Improvement Laboratory, Beltsville, MD 20705, 3

Department of 

Animal and Food Sciences, University of Delaware, Newark, DE 19717 

See abstract on page 37.

Structural and functional characterization of chicken IL-15, IL-16 and IL-17 

cDNAs. 

Min, Wongi, and Lillehoj, Hyun S. 

USDA, ARS, Animal Parasite Diseases Laboratory, ANRI, Beltsville, MD, 20705 

Limited progress in chicken cytokine immunobiology has slowed down immunological 

studies in poultry diseases. From our recently developed chicken intestinal gene database 

which contains >38,000 expressed sequence tags (EST), we searched for novel genes 

which are associated with innate and acquired immunities. This report describes the 

cloning and characterization of cDNAs encoding IL-15, IL-16 and IL-17 from our 

chicken intestinal library. DNA sequence analysis of a chicken IL-15 cDNA identified a 

187 amino acid open reading frame encoding a protein with a predicted molecular weight 

of 21,964, 2 potential N-linked glycosylation sites, 4 highly conserved Cys residues, 2 

out-of-frame AUG initiation codons in the 5’ untranslated region, and an unusually long 

(66 amino acid) signal peptide such that the expected size of the mature protein is 14,462 

Daltons. Chicken IL-15 and IL-2 were compared with regards to their molecular, 

cellular, and functional characteristics. The predicted amino acid sequences of both 

chicken cytokines showed greater homologies with mammalian IL-15s compared with 

mammalian IL-2s. Northern hybridization and RT-PCR demonstrated chicken IL-15 

gene transcripts in a wide variety of tissues and cell types while the chicken IL-2 gene 

was expressed only in concanavalin A-activated spleen cells. Both recombinant 

cytokines stimulated the growth of spleen T cells and enhanced the activity of natural 

killer cells in vitro. Subcutaneous injection with an expression plasmid encoding IL-15 

increased the percentage of CD3+ spleen T lymphocytes whereas injection of an IL-2 

cDNA augmented CD3+, CD4+, CD8+, TCR1+, and TCR2+ T cells. Collectively, these 

results indicate that chicken IL-15 and IL-2 are T cell growth factors potentially capable 

of enhancing cell-mediated immunity in vivo. 

IL-16 and IL-17 are proinflammatory cytokines produced by activated lymphocyte cells. 

Chicken IL-16 gene contained the entire open reading frame (ORF) of pro-IL-16 and 

encoded the mature IL-16 protein which consists of 607 amino acids. Chicken IL-16 

showed 86% sequence homology to duck pro-IL-16 and 49-52% to various mammalian 

IL-16s. By Northern blot analysis, IL-16 transcripts showed a restricted expression to the 

lymphoid tissues. Recombinant chicken IL-16 consisting of 149 C-terminal amino acids 

of pro-IL-16 was biologically active when expressed in COS-7 cells and showed 

chemoattractant property for lymphocytes. IL-17 contained a 507 bp open reading frame 

predicted to encode a protein of 169 amino acids with a molecular mass of 18.9 kDa. 

2003 ARS Immunology Research Worskshop 77 
Chicken IL-17 shared 37%-46% amino acid sequence identity with mammalian IL-17 

and homologous to the ORF 13 of Herpesvirus saimiri. By Northen blot analysis, IL-17 

transcripts were identified in a reticuloendotheliosis virus-transformed chicken 

lymphoblast cell line (CU205) and Con A-stimulated splenic lymphocytes, but not Cu91, 

RP9, RP13 cell lines or kidney, bursa, heart, spleen, cecal tonsils and thymus. 

Conditioned medium from COS-7 cells transfected with chicken IL-17 cDNA induced 

the production of IL-6 by embryonic fibroblasts. These studies will facilitate our 

understanding of poultry immune system. 

Differential pro-inflammatory cytokine mRNA expression in heterophils 

isolated from Salmonella-resistant and –susceptible chickens

Christina L. Swaggerty1, Pamela J. Ferro2, Lisa Rothwell3, Igal Y. Pevzner4, Michael H.

Kogut1

, and Pete Kaiser3 

1

 USDA, ARS, SPARC, 2881 F&B Road, College Station, TX 77845 2 Texas A&M 

University, College Station, TX 77843-2472 3

 Institute for Animal Health, Compton, 

Berkshire, RG20, 7NN, UK 4

 Cobb – Vantress, Inc., Siloam Springs, AR 72761-1030 

We have conducted studies utilizing a parental pair of broiler chickens (lines A and B) 

and the F1 reciprocal crosses (C and D). Previously we showed increased in vitro

heterophil functional efficiency translates to increased in vivo resistance against a 

systemic Salmonella enteritidis (SE) infection. Heterophils are capable of producing 

cytokines and are known to modulate acute protection against Salmonella in young 

poultry. We hypothesize that heterophils from SE-resistant chickens (A and D) have an 

upregulated pro-inflammatory cytokine response compared to heterophils from SEsusceptible chickens (B and C). In this study, heterophils were isolated from day-old 

chicks, stimulated with SE or SE opsonized with either normal chicken serum or immune 

serum against SE, and quantitative real-time RT-PCR using TaqMan chemistry was used 

to ascertain levels of cytokine mRNA expression. Heterophils from SE-resistant chicks 

(A and D) had significantly higher mRNA expression levels of pro-inflammatory 

cytokines (IL-6, IL-8, and IL-18) upon treatment with all stimulants compared to 

heterophils from SE-susceptible lines (B and C). Further, heterophils from SE-resistant 

chicks had significantly decreased mRNA expression levels of TGF-$4, an antiinflammatory cytokine, compared to heterophils from SE-susceptible chicks. These data 

indicate that chickens with heterophils more able to mount a pro-inflammatory cytokine 

response may determine if the chickens are going to be resistant or susceptible to 

Salmonella infections. Therefore heterophil functional efficiency and cytokine 

production may be useful biomarkers for poultry breeders to consider when developing 

new immunocompetent lines of birds. 

2003 ARS Immunology Research Worskshop 78 
Immunointervention Strategies 

Immunopotentiation of a recombinant adenovirus-FMD vaccine by IFNα in 

swine

Sonia A. Botton, Mario C. S. Brum, Elida M.Bautista, Marla J. Koster, William T. Golde, 

Marvin J. Grubman*. 

USDA, ARS, Plum Island Animal Disease Center, PO Box 848, Greenport, NY 11944-

0848. 

 *e-mail address: mgrubman@piadc.ars.usda.gov 

The adjuvant effect of interferon alpha (IFNα) was examined in swine vaccinated with a 

recombinant replication-defective adenovirus containing foot-and-mouth disease virus 

(FMDV) capsid (P1-2A) and 3C proteinase coding regions (Ad5-A24). Swine were 

divided into 5 groups and inoculated with high (5X109 PFU) or low (5X108 PFU) doses 

of Ad5-A24 in the presence or absence of porcine IFNα (Ad5pIFNα, 109 PFU). Control 

animals received 6X109

 PFU of adenovirus containing the glycoprotein from vesicular 

stomatitis virus (VSV) (Ad5VSV-G). All swine were challenged at 42 days post 

vaccination (dpv) with FMDV-A24. Prior to challenge, blood samples were examined for 

IFN production, induction of IFN-induced genes and FMDV-specific neutralizing 

antibodies (PRN70). After challenge a number of disease parameters were analyzed 

including clinical score, viremia and antibodies against viral structural and nonstructural 

(NS) proteins. 

Preliminary results indicate that both high-dose Ad5-A24 inoculated groups developed 

significant PRN70 by 14-21 dpv which was maintained until the day of challenge. While 

both low-dose inoculated groups had lower PRN70, the IFN group had an enhanced 

response. After challenge all control animals developed early viremia, vesicular lesions, 

and antibodies to FMDV NS proteins and high-levels of PRN70. Animals receiving low 

dose Ad5-A24 with no IFN had similar clinical signs, except that fewer animals 

developed viremia. In contrast pigs inoculated with the same dose plus IFN had a delayed 

onset of vesicular lesions and only one animal had detectable viremia. Animals 

vaccinated with high dose Ad5-A24 without IFN had no viremia, significantly fewer 

lesions and delayed onset of disease compared to the group given a low dose and no IFN. 

Four of five pigs vaccinated with high dose plus IFN were completely protected from

disease and only one animal in this group had a mild vesicular lesion restricted to the site 

of inoculation of challenge virus. Our results indicate that IFNα enhances the level of 

protection induced by the adenovirus-FMD vaccine against homologous FMDV, 

supporting the use of IFNα as a potential adjuvant in FMD vaccination strategies. 

2003 ARS Immunology Research Worskshop 79 
Swine vaccine trials with replication-defective human adenovirus serotype 5 

containing the capsid and 3C proteinase coding regions of Foot-and-Mouth

Disease Virus O1 Campos in the presence or absence Ad5-porcineGM-CSF

Luizinho Caron, Mario C.S. Brum, Mauro P. Moraes, Clarice W. Arns, William T. Golde 

& Marvin J. Grubman*. 

USDA, ARS, Plum Island Animal Disease Center, PO Box 848, Greenport, NY 11944-

0848. 

 *e-mail address: mgrubman@piadc.ars.usda.gov 

Granulocyte-macrophage colony stimulating factor (GM-CSF) has been widely used to 

successfully stimulate the immune response in vaccine trials involving HIV, hepatitis C 

and B, cancer etc. Because of these promising results, we constructed a recombinantdefective human adenovirus 5 (Ad5) containing porcine GM-CSF gene (Ad5-pGMCSF) 

to examine its effect on an experimental FMDV serotype O1 Campos vaccine that is 

poorly immunogenic. The recombinant virus was grown in 293 cells, cesium chloride 

purified and expression verified in IBRS2 cells by radio-immunoprecipitation and 

Western blotting. The activity of pGM-CSF expressed in IBRS2 cells was demonstrated 

in TF-1 cells, a cell line dependent on GM-CSF for growth. 

Eighteen pigs were divided into 5 groups: Groups 1 and 2, containing 3 animals each, 

were vaccinated with control virus, Ad5-Blue, or Ad5-pGMCSF and three other groups, 

4 animals per group, were vaccinated with Ad5 containing the capsid (P1-2A) and 3C 

proteinase coding regions of FMDV O1 Campos (Ad5-O1C) in the absence or presence 

of 2 different amounts of Ad5-pGMCSF. All animals were challenged 21 days postvaccination with a field strain of homologous virus. The results indicate that the presence 

of pGMCSF induced a more rapid FMDV-specific neutralizing antibody response but the 

titers declined faster then in the group given vaccine alone. Furthermore, after challenge, 

the groups administered both Ad5-pGMCSF and Ad5-O1C developed somewhat more 

severe disease when compared with the group given only Ad5-O1C. 

Development of an oral vaccine for Escherichia coli o157:h7

Evelyn A. Dean-Nystrom 

USDA, ARS, National Animal Disease Center, Ames, IA 

See abstract on page 39.

Oral dosing of mice with Escherichia coli expressing recombinant interferon 

gamma reduces Cryptosporidium parvum infection. 

James A. Harp*, Joan C. Oesper, Thomas A. Casey 

National Animal Disease Center, USDA-ARS, Ames, Iowa 50010. 

Cryptosporidium parvum is a protozoan parasite that causes intestinal infection in 

numerous mammalian species. Previous studies indicate that interferon gamma is critical 

to resistance to and recovery from C. parvum infection. We electroporated a gene for 

recombinant murine interferon gamma (rIFN) into a nontoxigenic E. coli strain that 

expresses the F41 adhesion pilus, allowing it to adhere in the mammalian small intestine 

(strain 4907.2.1). Groups of mice were treated at 1 day of age with an oral dose of 

2003 ARS Immunology Research Worskshop 80 
4907.2.1 expressing rIFN, either alone, or in combination with Lactobacillus brevis 

and/or killed C. parvum. Controls received phosphate buffered saline. All mice were 

orally inoculated with 103 viable C. parvum oocysts at 7 days of age. Mice were killed at 

2 weeks of age, and colon contents examined microscopically for the presence of C. 

parvum oocysts. The number of oocysts seen per microscopic field was compared 

between groups. Control mice had a mean of 2.1 oocysts per field. Mice receiving L 

.brevis, killed C. parvum, and 4907.2.1 had a mean of 1.75 oocysts per field. Mice 

receiving killed C. parvum and 4907.2.1 had a mean of 1.3 oocysts per field. Mice 

receiving 4907.2.1 alone had a mean of 1 oocyst per field. While these differences were 

not statistically significant, there was a clear trend that mice receiving E. coli expressing 

rIFN were less heavily infected with C. parvum. 

Stimulation of chicken leukocytes and reduction of Salmonella organ invasion 

in neonatal chicken by immunostimulatory CpG-oligodeoxynucleotide. 

Haiqi He and Michael H. Kogut 

USDA, ARS, Southern Plains Agricultural Research Center, College Station, TX77845 

See abstract on page 41.

Development of an inactivated Newcastle disease virosome vaccine that 

protects against challenge from velogenic Texas GB

Darrell R. Kapczynski and Terrence M. Tumpey 

USDA-ARS, Southeast Poultry Research Laboratory, Athens, GA 30605-2195 

While outbreaks of highly virulent Newcastle disease virus (NDV) are a major concern to 

the poultry industry internationally, economic losses caused by low virulent (lentogenic) 

strains continue to result from decreased egg production in layers and airsacculitis in 

broilers. Since lentogenic strains of NDV are widely used as vaccine strains, it is possible 

that these viruses, may be directly or indirectly responsible for some of the economic 

losses. In an effort to examine protection against NDV, we developed a non-replicating 

virosome vaccine that retains the ability to hemagglutinate chicken red blood cells and 

fuse with chicken embryo cells. Preliminary vaccine efficacy studies indicate that two 

intranasal doses containing 10 :g of virosomes protects chickens against lethal challenge 

from velogenic Texas GB. 

Development of immunotherapeutic recombinant chicken scFv antibodies 

reactive with an apical antigens of Eimeria. 

Lillehoj

1

, Hyun, Min1, Wongi and Kim

2

, Jin K. 

1

USDA, ARS, Animal Parasite Diseases Laboratory, ANRI, Beltsville, MD; 2Department 

of Microbiology, Changwon National University, Changwon, Korea. 

Slow progress in the identification of protective Eimeria antigens hinders the 

development of control strategies against avian coccidiosis. In order to identify the 

epitopes of Eimeria which are recognized by chicken B cells, novel immunization 

strategy was used that led to the development of several chicken monoclonal antibodies 

which blocked sporozoite invasion of host cells. While most antibodies recognized 

uncharacterized parasite surface proteins, two were identified with reactivity against 

2003 ARS Immunology Research Worskshop 81 
apical complex proteins located at the anterior tip of sporozoites. To express high 

concentration of these antibodies for potential application as immunotherapeutic reagents, 

a single chain fragment variable (ScFv) region gene was constructed by PCR 

amplification of the variable heavy (VH) and light (VL) chain genes with a flexible 

linker. By indirect immunofluorescence, recombinant ScFv antibody expressed in E. coli

showed identical binding specificity as the original monoclonal antibody against E. 

acervulina and E. tenella sporozoites, staining the apical complex and thus demonstrating 

reactivity against native parasite protein. These results demonstrate that recombinant 

6D12 ScFv recognizes the native conoid antigen on Eimeria. Potential application of this 

antibody in prophylactic therapy against avian coccidiosis is being explored using various 

delivery strategies including embryo injection. 

Adjuvant Effect of IL-1β, IL-2, IL-8, IL-15, IFN-α, IFN-γ, TGF-β4 and 

lymphotactin on DNA vaccination against Eimeria acervulina.

Wongi Min1

, Hyun S. Lillehoj

1

, and Peter Staeheli2. 1

 USDA, ARS, Animal Parasitic Diseases Laboratory, Animal and Natural Resources 

Institute, Building 1040, BARC-East, Beltsville, MD 20705 USA. 2

Department of 

Virology, University of Freiburg, D-79008, Freiburg, Germany. 

See abstract on page 40.

Comparison of immunologic responses to Brucella and Mycobacteruim

vaccines across species 

S.C. Olsen, A. Maue, M.V. Palmer, W.S. Stoffegren, and W.R. Waters. 

USDA, ARS, Bacterial Diseases of Livestock Research Unit, National Animal Disease 

Center, Ames, IA 50010. 

Eradication of brucellosis and tuberculosis from livestock populations is a high priority 

for regulatory personnel. Persistence of these diseases in wildlife reservoirs pose a 

substantial risk for transmission to livestock. In a series of studies conducted at the 

National Animal Disease Center, immunologic responses of wildlife species of interest to 

brucellosis and/or mycobacterium vaccines were characterized and compared to 

responses of cattle. As cell-mediated immunity is critical for long-term protection against 

diseases caused by intracellular bacteria, our studies particularly focused on proliferative 

reponses of peripheral blood mononuclear cells (PBMC), nitric oxide production, (-

interferon production, and flow cytometric analysis of proliferating PBMC subsets. Our 

data suggest that immunologic responses to Brucella or Mycobacterium vaccines may 

differ by species. Species that are considered to be closely related may demonstrate 

significant differences in immunologic responses to vaccination. Our data also suggest 

that immunologic responses to vaccination cannot be predicted but must be evaluated in 

the species of interest. Lastly, some wildlife species may have value as models for 

characterizing the role of humoral responses in ruminants as their immunologic responses 

to Brucella and Mycobacterium vaccines include robust antibody and deficient cellmediated responses. 

Does antigenic drift occur with avian influenza in poultry?

David Suarez and Chang-Won Lee 

2003 ARS Immunology Research Worskshop 82 
USDA, ARS, Southeast Poultry Research Laboratory, Athens, GA 30605-2195 USA 

Avian influenza in poultry and human influenza are both type A influenza viruses and 

both originated from an influenza reservoir found in wild birds, including ducks and 

gulls. Human influenza has been well characterized for antigenic drift, which is the 

accumulation of point mutations in the viral genome. These changes in the 

hemagglutinin gene can slowly create antibody escape mutants that make current 

vaccines less effective. Therefore the strains of virus in these killed vaccines must be 

updated on a yearly basis to provide optimal protection against the prevailing strains of 

circulating virus. In general when the predicted circulating strain of virus elicits an 

antibody titer with a fourfold loss in hemagglutination inhibition (HI) cross neutralization 

activity to the current vaccine, then the vaccine is changed. The changes in nucleotide 

similarity between the old and new vaccine strains are often less than 2%. For highly 

pathogenic avian influenza (HPAI), antigenic drift as it affects vaccine selection has not 

been a consideration in the past. Vaccine strains with differences of over 15% to the 

challenge strain can still provide good protection from clinical disease. There is a 

correlation with nucleotide sequence similarity and viral shed, with the closer a challenge 

strain is to the vaccine the greater reduction in viral shed. The disconnect between 

human influenza and HPAI vaccines may be that human influenza is a mucosal infection, 

but HPAI is a mucosal and a systemic infection. Vaccines for influenza do not prevent 

infection, but they do reduce viral shed and prevent or lessen clinical signs. Antibodies in 

the blood stream may effectively prevent the systemic phase of disease with HPAI 

viruses. However, recent studies of low pathogenic avian influenza from Mexico and the 

U.S. may present a different view of antigenic drift in poultry. In Mexico H5N2 LPAI has 

circulated for 9 years and vaccination with two different vaccines have been used widely. 

When recent isolates are examined by HI cross neutralization to the killed vaccine, large 

decreases in titers are observed. However, when examining H7N2 LPAI viruses that have 

been circulating in the U.S. since 1994, large decreases in HI cross neutralization titers 

are not observed. In both cases nucleotide sequence differences between 3-5% are 

observed. One major difference in the two groups of isolates is whether vaccination is

used or not. The possible effect that antigenic drift could have on vaccination practices 

will be discussed. 

2003 ARS Immunology Research Worskshop 83 
Recombinant Paramyxovirus type 1-Avian Influenza-H7 virus as a vaccine 

for protection of chickens against influenza and Newcastle disease

D.E. Swayne

1

, D.L. Suarez

1

, S. Schultz-Cherry1, T.M. Tumpey 

1

, D.J. King1, T. Nakaya2, 

P. Palese2

 and A. Garcia-Sastre2

1

USDA, ARS, Southeast Poultry Research Laboratory, Athens, GA 30605-2195 USA 

2

Department of Microbiology, Mount Sinai School of Medicine, New York, NY 10029 

USA 

Current vaccines to prevent avian influenza rely upon labor-intensive parenteral injection. 

A more advantageous vaccine would be capable of administration by mass immunization 

methods such as spray or water vaccination. A recombinant vaccine (rNDV-AIV-H7) 

was constructed by using a lentogenic Paramyxovirus type 1 vector (Newcastle disease 

virus [NDV] B1 strain) with insertion of the hemagglutinin (HA) gene from avian 

influenza virus (AIV) A/chicken/NY/13142-5/94 (H7N2). The recombinant virus had 

stable insertion and expression of the H7 AIV HA gene as evident by detection of HA 

expression via immunofluorescence in infected Vero cells. The rNDV-AIV-H7 replicated 

in 9-10 day embryonating chicken eggs and exhibited hemagglutinating activity from

both NDV and AI proteins that was inhibited by antisera against both NDV and AIV H7. 

Groups of 2-week-old white Leghorn chickens were vaccinated with transfectant NDV 

vector (tNDV), rNDV-AIV-H7 or sterile allantoic fluid and were challenged 2 weeks 

later with viscerotropic velogenic NDV (vvNDV) or highly pathogenic (HP) AIV. The 

sham-vaccinated birds were not protected from vvNDV or HP AIV challenge. The 

transfectant NDV vaccine provided 70% protection for NDV challenge but did not 

protect against AIV challenge. The rNDV-AIV-H7 vaccine provided partial protection 

(40%) from vvNDV and HP AIV challenge. The serologic response was examined in 

chickens that received one or two immunizations of the rNDV-AIV-H7 vaccine. Based 

on hemagglutination inhibition and ELISA tests, chickens that received a vaccine boost 

seroconverted to AIV H7, but the serologic response was weak in birds that received only 

one vaccination. This demonstrates the potential for NDV for use as a vaccine vector in 

expressing AIV proteins. 

Evaluation of a commercial avian influenza (H7N2) vaccine for protection in 

turkeys against an avian influenza virus (H7N2) isolated from turkeys in 

Virginia during 2002

Terrence M. Tumpey, Darrell R. Kapczynski, Rashida Horton and David E. Swayne

Southeast Poultry Research Laboratory, USDA, Agricultural Research Service, Athens, 

GA 30605. 

During the spring of 2002, a low pathogenic avian influenza (LPAI) A (H7N2) virus 

caused a major outbreak in commercial poultry in Virginia and adjacent states. The low 

pathogenic avian influenza (LPAI) virus primarily affected turkey flocks causing 

respiratory distress and decreased egg production. Experimentally, turkeys were more

susceptible than chickens to H7N2 virus infection and higher titers of infectious virus 

could be recovered from the oropharynx in comparison to the cloaca. The outbreak of 

H7N2 virus raised concerns regarding the availability of vaccines that could be used for 

the prevention and control of this virus in poultry. We sought to determine if an existing 

2003 ARS Immunology Research Worskshop 84 
commercial AI vaccine prepared from a 1997 seed stock virus could provide protection 

against a 2002 LPAI H7N2 virus isolated from a turkey (A/Turkey/Virgina/158512/02 

[TV/02]) in Virginia that was from the same lineage as the vaccine virus. The inactivated 

AI vaccine, prepared from A/Chicken/Pennslyvania/21342/97 (CP/97) virus, significantly 

reduced viral shedding from vaccinated turkeys in comparison to sham controls, but did 

not prevent infection. The protective effect of vaccination correlated with the level of

virus-specific antibody as a second dose of vaccine increased antiviral serum IgG and 

hemagglutination inhibition (HI) reactivity titers in two different turkey age groups. 

Serum from CP/97-vaccinated turkeys reacted equally well to CP/97 and TV/02 antigens 

by HI and ELISA. These results demonstrate the potential benefit of using an 

antigenicly-related 1997 H7N2 virus as a vaccine candidate for protection in poultry 

against a H7N2 virus isolate from 2002. 

Antigen-induced IFN-γ, nitric oxide, and TNF-α production by blood 

mononuclear cells from Mycobacterium bovis-infected cattle: diagnostic 

implications.

W. R. Waters

1

, M. V. Palmer

1

, D. L. Whipple1, M. P. Carlson2, and B. J. Nonnecke1

1

 USDA, ARS, National Animal Disease Center, Ames, IA 50010; 2 University of 

Nebraska Veterinary Diagnostic Center, Toxicology Laboratory, Lincoln, NE 68583-

0907 

Bovine tuberculosis in the United States has proven costly to cattle producers as well as 

government regulatory agencies. While in vivo responsiveness to mycobacterial antigens 

is the current standard for the diagnosis of tuberculosis, in vitro assays are gaining 

acceptance, especially as ancillary or complimentary tests. To evaluate in vitro indices of 

cellular sensitization, antigen induced interferon (IFN)-γ, nitric oxide (NO), and tumor 

necrosis factor (TNF)-α responses by blood mononuclear cells from Mycobacterium 

bovis-infected cattle were quantified and compared. Using an aerosol model of infection, 

two doses each of two strains of M. bovis (95-1315 and HC-2045T) were used to induce a 

range of IFN-γ, NO, and TNF-α responses. Infection specific increases in NO, but not 

IFN-γ or TNF-α, were detected in nonstimulated cultures at 48h, indicative of 

nonspecific activation and spontaneous release of NO. The infective dose of M. bovis 

also influenced responses. At 34d postinfection, IFN-γ, NO, and TNF-α responses in 

antigen-stimulated cells from cattle receiving 105 cfu M. bovis were greater than 

responses of cells from cattle infected with 103

 cfu M. bovis. The NO response, but not 

IFN-γ and TNF-α responses, was influenced by infective strain of M. bovis. TNF-α, NO, 

and IFN-γ responses followed similar kinetics with strong positive associations between 

the three readouts. Overall, these findings indicate that NO and TNF-α, like IFN-γ, may 

prove useful as indices for the diagnosis of bovine tuberculosis. 

2003 ARS Immunology Research Worskshop 85 
Working Group Reports 

Immunity and Disease Resistance 

Immunointervention 

Immunity and the Pathogenesis of Diseases 

Nutrition and Immune Function Interations 

2003 ARS Immunology Research Worskshop 86 
Immunity and Disease Resistance 

 

Working Group Leaders:

• Lanie Bilodeau, Immunologists, Catfish Genetics Research Unit, ARS, 

Stoneville, MS 

• Lou Gasbarre, Research Leader, Bovine Functional Genomics Research Unit, Animal 

and Natural Resources Institute, ARS, Beltsville, MD 

• Michael H. Kogut, Immunologist, Southern Plains Agricultural Research Center, 

ARS, College Station, TX 

• Joan Lunney, Immunologist, Animal Parasite Diseases Laboratory, Animal and 

Natural Resources Institute, ARS, Beltsville, MD

Research Priority 1

Identification of genes defining host disease resistance 

1. Background: There is a need for identification and characterization of genes that 

confer host resistance to important production and foodborne diseases. Studies must 

determine whether it is possible to identify food animal species that have the potential to 

be “healthier.”

2. Issue: Identification and characterization of genes defining host disease resistance will 

enable producers to reduce disease incidence using genomic approaches. Alternately, 

identifying disease resistant animals may reveal new approaches for disease control. 

3. Research Objectives: 

A Identify/develop commercial/experimental animal resource populations for 

identification and mapping of immunologically relevant genes 

B. Identify and characterize host genes that control important immunologic 

responses; determine whether alleles of certain genes would enhance animal 

health 

C. Define important genetic polymorphisms that affect host resistance/susceptibility 

D. Define gene expression patterns and regulatory phenomena that affect the level of 

host resistance 

E. Utilize genomic information to reveal new approaches for disease control or 

biotherapeutic production 

4. Proposed Accomplishments:

A. Mapping projects will identify informative populations for disease studies. These 

can then be linked with known populations and commercial breeding stock. 

Information gained can be used to plan modification of populations for specific 

production/health issues 

B. Information will be developed on correlation of disease resistance phenotype with 

desired production traits by phenotyping for disease resistance traits using high 

throughput (HTS) immune and disease screens coordinated with production 

parameters. This will prove relevance for utilization of disease resistance in 

commercial populations/genomic stock 

2003 ARS Immunology Research Worskshop 87 
C. Expansion of gene expression/regulation approaches, includes microarray 

[targeted and full genomic], real-time RT-PCR, proteomic studies, RNAi 

technology, leading to a more thorough understanding of disease resistance 

mechanisms

D. Expanded knowledge of targeted genomic areas, Immunoglobulin (Ig), T cell 

receptor (TCR), major histocompatibility (MHC), pattern recognition receptors 

(PRR), Toll-like receptor (TLR) pathways, and their impact on health and disease 

5. Proposed Collaborations: Work with scientists in NP101 and NP106 programs to 

utilize genomics tools developed; similarly; work with national (NRSP008) and 

international genome efforts, e.g. sequencing and microarray projects; work with national 

swine PRRS initiative to target specific disease. 

6. Anticipated project duration: Significant progress should be made across taxa for 

gene identification/characterization studies within 5 years.

7. Impact: 

A. Markers/genes regulating disease resistance will be identified. This information 

will be used for developing allele tests, e.g., single nucleotide polymorphisms 

(SNPs) for fast, effective identification of resistant animals; for marker assisted 

selection (MAS) approaches to increase frequency in target populations; or for 

development of transgenic or clone lines of disease resistant animals 

B. Definition of novel mechanisms of protective immunity. Apply information 

gained from genomics to developing new biotherapeutics, vaccine approaches 

(with APHIS). 

C. Develop approaches for diagnostics and new rationales for immune control of 

diseases 

Research Priority 2

Induction/regulation of immune responses through mucosal surfaces 

1. Background: Many infectious diseases enter through respiratory and gastrointestinal 

tissues and infect though mucosal surfaces. There is a need for a better understanding of 

what drives immune responses to infection/invasion of the host. Studies focused on 

host/pathogen interactions at mucosal surfaces are essential to define early events 

relevant to disease resistance. 

2. Issue: Identification of pathogen uptake sites/mechanisms will enhance understanding 

of immune responses. Pathogen entry occurs largely across mucosal membranes. In 

addition to uptake, the study of induction/regulation of immune responses through 

mucosal surfaces are needed

3. Research Objectives: 

A. Define cell populations/subsets that affect resistance at mucosal surfaces; 

determine how neonatal development affects these populations. 

B. Define gene expression patterns and regulatory phenomena that affect the level of 

host resistance at mucosal surfaces. 

C. Use information from microbial genomics to define substances from pathogens 

that stimulate/regulate immune responses at mucosal surfaces. 

D. Define the affects of stress/diet/age on immune responses at mucosal surfaces. 

2003 ARS Immunology Research Worskshop 88 
4. Proposed Accomplishments:

A. Identification of regional (respiratory, intestinal, reproduction, skin) areas of 

infection/points of uptake; Determination of mucosal cell populations involved in 

disease processes; Development of immune tools to assess immune cell subsets 

regulating responses; Determination of age-related development associations with 

infection; Development of gene expression/regulation approaches and reagents to 

quantitate chemokines and cytokines. 

B. Determine relationship of infection kinetics (in blood/specific mucosal tissues) 

and mucosal immune function; Identification of relevant genes controlling 

infection processes, e.g., TLR, anti-microbial peptides; MHC genes involved. 

C. Application of microbial genomics information to probe critical microbial genes 

influencing host disease response and means, or mechanisms, of pathogen 

invasion/evasion/infection. 

5. Proposed Collaborations: Interactions with university scientists through NC projects; 

with international researchers through UK BBSRC, Spain INIA; with commercial 

producers/animal health companies 

6. Anticipated project duration: Significant progress should be made across taxa for 

gene identification/characterization studies within 5 years.

7. Impact: 

A. Definition of local protective defense mechanisms; Definition of pathways for 

interventions, for development of both traditional and therapeutic vaccines, and 

for preparation of novel mucosally targeted biotherapeutics 

B. Improved vaccine efficacy; verification of efficacy of mucosal delivery vectors 

and adjuvants. 

C. (with APHIS) Develop approaches for diagnostics and rationale for immune 

control of diseases 

Research Priority 3

Induction/regulation of innate responses; influence/regulation of 

acquired immunity 

1. Background: The role of innate immunity, and its influence on acquired immunity, for 

disease control are still undefined for most agricultural species. Research in this area will 

influence development of treatments/vaccines/therapeutics, and improve management 

practices. 

2. Issue: The induction/regulation of innate immunity affects disease kinetics, disease 

resistance, and ultimately animal production. Determination of induction/regulation 

mechanisms of innate responses, and their influence on the effectiveness of acquired and 

protective anti-disease immunity, is needed. 

3. Research Objectives: 

A. Define cell populations/subsets that affect innate immunity. 

B. Define gene expression patterns and regulatory phenomena that affect the level of 

host innate immunity. 

C. Use information from microbial genomics to define substances from pathogens 

that stimulate/regulate innate immune responses; identify the evolutionarily 

conserved molecular constituents (PAMPs) of infectious microbes. 

D. Define the affects of stress/diet/age on innate immune responses 

2003 ARS Immunology Research Worskshop 89 
4. Proposed Accomplishments: 

A. Knowledge of genomic organization of loci involved in the germ-line encoded 

pattern recognition receptors (PRR), e.g., TLRs, mannose receptors, scavenger 

receptors, and complement receptors. Expression levels of PRR on effector cells 

of innate immunity (macrophages, NK cells, granulocytes, epithelial). 

B. Identification of pathogen-associated molecular patterns (PAMPs) and their 

interaction with PRR will inform pathogen control measures and help identify 

new biotherapeutics. 

C. Effect of innate responses on intensity and bias of pathogen-specific effector 

acquired immune responses, and activation of the co-stimulatory molecules 

required for T cell responses; determine role of innate and acquired immunity in 

disease kinetics. 

D. Knowledge of development/regulation of immunity in neonates/early 

developmental stages 

E. Effect of stress and nutritional status on innate immune function. 

5. Proposed Collaborations: Interactions with university scientists through NC projects; 

with international researchers through UK BBSRC, Spain INIA; with commercial 

producers/animal health companies

6. Anticipated project duration: Significant progress should be made across taxa for 

gene identification/characterization studies within 5 years. 

7. Impact: Identification of animals with broader spectrum disease resistance 

Development of Biotherapeutics; Definition of Local protective defense mechanisms; 

Identification of innate immune pathways for interventions/traditional and therapeutic 

vaccines and for development of new adjuvants and improvement of vaccine efficacy 

Research Priority 4

Bioinformatics 

1. Background: Improvements in genomics technologies have led to high-throughput 

sequencing which generates large data sets that require bioinformatics tools and 

computing power necessary for analysis. Currently, the infrastructure does not exist for 

handling agency-wide, or in some cases unit-level, collation of data and extensive 

analyses. These resources need to be developed at the unit, area, agency and national 

levels. 

2. Issue: Large volumes of data are generated that are difficult to analyze. For this reason, 

bioinformatics and genomics resources need to be developed and integrated into ARS 

programs and research units. 

3. Research Objective: 

A Develop standardized bioinformatics tools and resources that can be utilized by 

multiple programs and research units. 

B. Develop tools/databases for species sequence, data mining 

4. Proposed Accomplishments:

A. Improve high-powered computer support and information exchange/accessibility. 

Tools/Statistical analyses of disease mapping info that adjust to population 

structure and enable efficient data management. Analysis tools and MAS 

approaches 

2003 ARS Immunology Research Worskshop 90 
B. Improved in silico study tools, e.g., comparative analysis tools; improved 

annotation tools 

D. Proteomics-based tools 

5. Proposed Collaborations: Nationally centered bioinformatics resources but species 

relevant NSRP-8 and TIGR as potential resources; Need for Long term funding for 

maintenance and updating of websites 

6. Anticipated project duration: Significant progress should be made across taxa for 

gene identification/characterization studies within 5 years. 

7. Impact: Utilize genome sequencing information (animals, microbes); Eliminate 

redundancy;Improved comparative bioinformatics; Improved information exchange. Tool 

availability for broader animal health community; In silico tools for efficient vaccine 

development

Research Priority 5

Enhance Immune Toolkit for major food animal species 

1. Background: Basic tools to measure details of immune responses in major food 

animal species are not available or critical components are missing. For example, there 

are insufficient panels of monoclonal antibodies (mAb) that identify and quantitate CD

antigens on immune cell surfaces, chemokines and cytokines; molecular probes and 

assays for immune markers; cloned and expressed bioactive chemokines and cytokines. 

Availability of quality targeted immune gene microarrays will be another important 

resource. 

2. Issue: Because of limited markets for these products, production of high quality 

reagents is often left to researchers; for some species commercial sources of these 

reagents are limited or non-existent.

3. Research Objectives: 

A. Establish national and international networks to develop priorities for toolkit needs 

for each species. 

B. Coordinate tool development so that priority reagents become available. Work 

with commercial sources to market products. Simultaneously work with 

repositories to have independent sources of hybridoma cell lines and expression 

vectors for cloned and expressed bioactive chemokines and cytokines so that 

researchers can produce large quantities for research efforts. 

4. Proposed Accomplishments: 

A. Panels of reagents, mAb, cloned and expressed bioactive chemokines and 

cytokines, immune gene microarrays, will be available to researchers for their 

disease resistance studies 

B. Critical control points in disease processes, and in immune events controlling 

these processes, will be revealed 

C. Technology transfer opportunities will be expanded 

5. Proposed Collaborations: Interactions with university scientists through NC projects; 

with international researchers through UK BBSRC, Spain INIA; with commercial 

producers/animal health companies. Need for long term funding for maintenance and 

updating of immune toolkit websites 

6. Anticipated project duration: Significant progress should be made across taxa for 

gene identification/characterization studies within 5 years.

2003 ARS Immunology Research Worskshop 91 
7. Impact: Effective tools to explore phenotyping animal immunity, to determine 

protective immune responses, to confirm bioactive therapeutics. Tools to quantitate 

immune responses, to validate models of immunity, to assess differential genetic 

responses of resistant/susceptible to infectious disease challenges 

Note: Specific disease targets will be based on stakeholders’ priorities. 

Research Priorities have been listed but numbers do not indicate relative priorities since 

all approaches are needed for maximal impact on improving disease resistance. 

2003 ARS Immunology Research Worskshop 92 
Immunointervention 

Working Group Leaders: • Marcus Kehrli, Research Leader, Virus and Prion Diseases Research Laboratory, 

National Animal Disease Center, ARS, Ames, IA

• Hyun Lillehoi, Immunologist, Animal Parasite Diseases Laboratory, Animal and 

Natural Resources Institute, ARS, Beltsville, MD

• David White, Veterinary Medical Officer, Arthropod-borne Animal Diseases 

Research Laboratory, ARS, Laramie, WY

Research Priority 1

Identify mechanisms of innate immunity and their regulation critical in 

control of animal diseases. 

1. Background: Many diseases are not amenable to control by traditional vaccination 

approaches because they are caused by an array of antigenically distinct opportunistic 

pathogens. Moreover, these opportunistic diseases are often associated with stress, 

broad-based immune impairment and/or overwhelming pathogen exposure. Some of 

these diseases may be better controlled by modulation of the innate immune system at 

critical growth/production stages to improve the host’s capacity to resist opportunistic 

infections and disease. 

2. Issue: There is a lack of knowledge about factors regulating humoral and cellular 

components of innate immunity. 

3. Research Objectives: 

A. Identify critical components of innate immunity in cattle, pigs and poultry and 

how they are regulated.

B. Develop effective strategies to evaluate the innate immune response in domestic 

livestock and poultry, including neonatal and embryonic immunity, innate 

immune cell function (neutrophils, NK cells, etc.), accessory cell function, and 

assay development for immunological reagents. 

C. Identify inter-relationships between innate immunity and adaptive immunity in 

cattle, pigs and poultry, and how innate immunity can impact host adaptive 

immune responses to vaccination or natural infection. 

4. Proposed Accomplishments:

A. Identification of various receptors and cell signaling pathways in livestock that 

trigger innate immunity. This should also offer insights on how to regulate the 

activation of innate immunity. 

B. More effective methods to evalute methods of immunointervention of the innate 

immune system. 

C. Identification of key regulatory relationships between the innate and adaptive 

immune systems. This will provide insight on what critical innate costimulatory factors are needed to generate an efficacious immune response. 

2003 ARS Immunology Research Worskshop 93 
5. Proposed Collaborations: Appropriate ARS CRIS projects, various university 

collaborations, and pharmaceutical and biologics companies. 

6. Anticipated project duration: 5+ years

7. Impact: 

A. Increased understanding of critical innate immunity components in livestock 

will provide a knowledge base that can be exploited to make science-based 

decisions to reduce disease incidence and severity through targeted 

immunointervention techniques. 

B. Increased ability to screen modulators of innate immunity against specific 

pathogens. 

C. Increased understanding of how innate immunity impacts vaccine responses in 

livestock will provide a knowledge base for better adjuvant and costimulatory 

molecule inclusion in vaccines, thus improving vaccine efficacy. 

Research Priority 2

Identify vaccines that elicit protective cell-mediated immunity 

1. Background: Traditional approaches to disease control have been through vaccination 

to generate protective antibody responses. Many diseases have been effectively 

controlled by this approach. However, most animal diseases that persist as problems in 

the U.S. today have not been amenable to control by these traditional approaches. One 

explanation of how these pathogens remain uncontrolled is the absence of an appropriate 

protective immune response. Some pathogens may evade the immune system through 

many mechanisms and may require generation of antigen-specific cytotoxic T-cells to be 

appropriately controlled. Identification of protective immune responses to the proper 

antigens requires immunological tools not currently available for widespread use in 

livestock or wildlife that are important for us to control. 

2. Issue: Traditional methods of assessing vaccine efficacy are to measure antibody 

responses and protection against disease challenge. Unfortunately antibody responses are 

not always predictive of a protective immune response against live pathogen challenge. 

We need better tools to assess cell-mediated immune responses in all livestock species. 

3. Research Objectives: 

A. Develop new methods in livestock for detecting and measuring cell-mediated 

immunity. 

B. Identify antigens critical for the development of cell-mediated immunity. 

C. Identify methods to regulate the development of cell-mediated immunity. 

D. Develop antigen delivery systems (expression vectors for various host animal 

species to be used for emerging diseases) that target immune system

compartments (mucosal, peripheral, reproductive tract and other privileged 

sites). 

E. Explore effects of modulation or elimination of T-regulatory cell subsets on 

induction of protective responses in domestic animals and poultry. Develop 

novel immunomodulators to suppress regulatory cell. 

4. Proposed Accomplishments:

A. Tools for measuring cell-mediated immunity will enable discovery, development 

and approval of new effective vaccines with cell-mediated immunity label 

claims. 

2003 ARS Immunology Research Worskshop 94 
B. Methods to predict and select appropriate pathogen antigens for incorporation 

into vaccines driving cell-mediated immune responses will enhance our response 

time in the event of emerging diseases. 

C. Understanding the regulatory factors that contribute to an effective cell-mediated 

immune response will allow us to create vaccines that do a better job of 

stimulating immunity than the pathogen does by itself. 

D. An improved capacity to rapidly develop vaccines against new and emerging 

pathogens. 

5. Proposed Collaborations:

6. Anticipated project duration: 5 years + 

7. Impact: 

A. Vaccines that aid in the control of diseases not currently effectively controlled. 

B. More rapid vaccine discovery and approval. 

C. Better methods to elicit and control immune responses requiring cell-mediated 

immunity. 

D. An increased capacity to rapidly respond to emerging disease outbreaks. 

Research Priority 3

Identify vaccines that elicit protective humoral immunity 

1. Background: Some pathogens elicit strong humoral immune responses that are not 

highly protective. The survival of these pathogens may depend on their ability to divert 

the efforts of the immune system towards antigens that do not facilitate their clearance 

from the host. 

2. Issue: There is a lack of basic knowledge in what drives successful humoral immune 

responses against certain pathogens. More indepth knowledge on regulation of immune 

responses in different compartments (e.g., respiratory tract, intestinal tract, reproductive 

tract, mammary gland) of the body is needed to better guide vaccine discovery and 

development into commercial products. 

3. Research Objectives: 

A. Identify factors regulating antibody responses to antigens. 

B. Develop computational methods to increase the efficiency of the design and 

testing of vaccines. 

C. Develop allied testing methods to distinguish between vaccinated and naturally 

infected animals. 

D. Develop broad spectrum, targeted delivery systems (target dendritic cells or

others) for immunomodulator and/or peptide and/or DNA delivery to effector 

cells. 

E. Elucidate factors affecting efficacy of current adjuvant systems in domestic 

animals (i.e. optmized CpG sequences for all livestock species) and develop new 

immunoadjuvants from novel sources (snake venoms, modified superantigens, 

probiotics as adjuvants, cytokines – IL-15, hormones, vitamins and trace 

minerals, carotenoids, phenolics, etc.). 

F. Delineate the specific conditions required for several different broad-based 

protein or nucleic acid delivery systems – i.e. needle-less systems, microgels, 

biodegradable depot systems, in ovo injector systems – while concentrating on 

important factors such as multiple use strategies, limiting the injected volume, 

2003 ARS Immunology Research Worskshop 95 
and livestock or wildlife behavioural responses (fear and/or aggression in 

response to noise or pain) to ensure field practicality. Evaluate natural delivery 

agents (i.e. viruses that cross the placenta naturally to deliver nucleic acids to the 

developing fetus). 

4. Proposed Accomplishments:

A. A better knowledge base of how antibody genes are selected and arranged for 

various classes of antigens, we can more effectively design next generation 

vaccines. 

B. By leveraging the knowledge database from human disease research, we will 

build a comparative immunology database that can be a guide to antigen 

selection for use in vaccines. 

C. Marked vaccines to distinguish vaccinates from natural infection. 

D. Adjuvant development (i.e. CpG) and/or delivery, peptide or DNA delivery (i.e. 

microgels, biodegradable depot systems). 

E. Adjuvants and immune modulators that target more effective immune responses. 

F. Broad-based delivery systems applicable to both livestock and wildlife that 

account for the practical realities of immunization in the field – especially in the 

case of wildlife, where very little information exists on the efficacy and 

reliability of strategies used in domestic livestock and poultry. 

5. Proposed Collaborations:

6. Anticipated project duration: 5 years + 

7. Impact: 

A. Vaccines that aid in the control of diseases not currently effectively controlled. 

B. More rapid vaccine discovery and approval. 

C. Marked vaccines will enable eradication programs once disease prevalence is 

below a certain economic threshold. 

D. By partnering with industry early in the antigen selection process we will better 

ensure viability of developed products. 

E. Effective strategies to immunize and modulate the responses of domestic animals 

and wildlife benefit agriculture by lesser manipulation of animals and lesser 

exposure to adjuvants that may impact meat or hide quality. 

F. Effective strategies to immunize wildlife will significantly decrease the risk of

disease in domestic animals because of a reduction in the size of the reservoir, 

both in the U.S. and abroad. 

Research Priority 4

Develop immunological reagents for domestic animals, poultry, and 

wildlife that are necessary to aid in the control of diseases we face today. 

1. Background: Although many significant advances have been made in the past 3 

decades, cutting-edge research into host immune responses in domestic animals and 

wildlife has been hampered by the cost of developing the necessary reagents and tools of 

immunological research. These include but are not limited to species-specific cytokines, 

effector molecules, and receptors, and monoclonal antibodies against each. These tools 

can be identified and made available with the necessary resources and investment to 

produce them. However, mechanisms need to be in place to allow production and 

sharing of these reagents at reasonable costs. 

2003 ARS Immunology Research Worskshop 96 
2. Issue: Reagents for the evaluation of immunological parameters in domestic animals 

and wildlife are severely lacking. Moreover, syngeneic animals or protocols for culture 

and preservation of immunological cells are necessary for many studies of basic 

immunological function. 

3. Research Objectives: 

A. Identify an appropriate infrastructure with the capacity to express various 

recombinant proteins and produce monoclonal antibodies against them. This 

may represent a government “subsidized” company that in turn recovers some of 

their manufacturing costs through the sale of these reagents to researchers 

around the world. 

B. Clone, express in native form, characterize, and develop antibodies to MHC 

Class I and II, acute phase proteins, complement proteins, cytokines, antibody 

isotypes, innate immune system receptors (TLR’s and others), effector 

molecules (i.e. perforin, granzyme, etc.), adhesion molecules, and cell surface 

markers of lymphocyte subsets for relevant domestic livestock and poultry 

species. House these reagents in some sort of centralized facility (either 

government or corporate) while balancing the free exchange of reagents for 

research purposes with the patent and intellectual property rights that will make 

them viable commercial products. Use the NIH Tetramer core facility as a 

model of a government-run facility for vital research reagents. 

C. Develop strategies to produce syngeneic domestic livestock strains (inbreeding) 

or alternatives to fully syngeneic animal development through in vitro 

manipulation immune cells and/or developing MHC-matched animal lines. 

D. Improve production and reduce cost of protein or nucleic acid substrates to make 

these strategies viable to animal production systems through high efficiency, 

scalable fermentation or synthesis processes that will produce native (or at least 

bioactive) protein. 

4. Proposed Accomplishments:

A. A centrally-maintained research facility that provides high-quality reagents to 

researchers worldwide for the evaluation of immunological parameters in 

domestic livestock and poultry. 

B. A reasonably priced supply of new immunological reagents for use in livestock, 

poultry and wildlife research. 

5. Proposed Collaborations:

6. Anticipated project duration: 5 years+ (ongoing supplier) 

7. Impact: 

A. Availability of new immunological reagents will increase the number of 

laboratories capable of investigating host immune responses against disease. 

B. Increasing the number of laboratories investigating immunity against disease 

with modern immunological reagents and tools will increase the probability of 

identifying new and more efficacious means of immunointervention. 

C. New avenues of research not currently possible will be opened up for scientists to 

follow. 

2003 ARS Immunology Research Worskshop 97 
Research Priority 5

Disease challenge models are needed for many diseases affecting 

domestic animals and wildlife. 

1. Background: Appropriate disease challenge models will be needed to accurately 

predict the success of any method of immunointervention in livestock or wildlife. It is 

possible to not recognize successful methods of immunointervention in instances where 

the model has an overwhelming pathogen challenge not representative of the natural 

disease exposure. Animal disease researchers have the unique opportunity of working in 

the natural host for most diseases that are studied, but reliable evaluation of intervention 

strategies requires standardized models and conditions that allow for the delineation of 

intervention effects. Make protocols and strains of pathogens used in models freely 

available to any interested researcher. Use models to elucidate immune functions 

relevant to protection from infection or disease, duration of immunity from different 

intervention protocols (dose, route, formulation, and timing vs. generation of lifelong 

immunity), and biomarkers of relevant clinical endpoints for testing and future efficacy 

evaluation. Finally, use models to define immunogenic relationships within and between 

pathogens to allow for the development and evaluation of cross-protection between 

vaccine components or interference between immunomodulatory agents.

2. Issue: Models may fail to accurately predict the outcome of immunointervention 

strategies in the “real world”. 

3. Research Objectives: 

A. Develop challenge models for diseases of domestic animals and wildlife. 

B. Develop generalist in vivo tests of immune function that are predictive of disease 

outcome. 

4. Proposed Accomplishments:

A. Models of disease will help both in the development and continuing evaluation 

of immunointervention strategies. 

B. Biomarkers of immunointervention success will enhance our capacity to screen 

for the most efficacious method to elicit a protective immune response. 

5. Proposed Collaborations:

6. Anticipated project duration: 5 years 

7. Impact: 

A. More rapid and uniform testing of immunointervention strategies across 

laboratories. 

B. Biomarkers that may be used in large scale studies on efficacious 

immunointervention strategies will reduce the cost of identifying new disease 

control methods. 

Research Priority 6

Vaccine vigilance 

1. Background: There are consequences of vaccination that we need to assess to ensure 

we are not adversely affecting livestock performance and well-being. 

2. Issue: There is a need to investigate and characterize immunopathological 

mechanisms in domestic livestock and poultry. 

2003 ARS Immunology Research Worskshop 98 
3. Research Objectives: 

A. Characterize hyper-responsiveness of the immune system, as an allergic reaction 

(i.e. peanut allergy), an adverse response to immunointervention, or as a part of 

other physiopathological processes (i.e. pulmonary hypertension syndrome in 

chickens, autoimmune utiligo in chickens). 

B. Create a database of field response rates and adverse event rates for evaluation of 

current and future vaccine efficacy. 

C. Characterize field variation of important pathogens and assess its impact on 

vaccine failure. 

D. Develop strategies for inclusion of multiple strains of the same pathogen, or 

multiple pathogens, in a single vaccine dose while avoiding the problems of 

immunodominance/original antigenic sin, antigen overload, and induction of 

tolerance. 

E. Optimize use of existing vaccines and immunomodulators for intervention at 

embryogenesis, birth, weaning, or prior to stress (shipping, comingling, 

slaughter). 

F. Develop in vitro assays for efficacy and safety of immunointervention agents 

and/or insults to the immune system. 

4. Proposed Accomplishments:

A. Fewer adverse event reports. 

B. Centralized database of vaccine usage and adverse events will lead to better 

monitoring of efficacy and failure. This database could be developed in 

conjunction with a broader database of “animal passports/ID”. 

C. Vaccines that are more producer-friendly through reduced animal manipulation 

while retaining the efficacy of individual preparations. Vaccines would also be 

of great use in wildlife management because of decreased need for contact and 

manipulation of wildlife. 

D. Ensure efficacy of modulation of relevant immunological endpoints in wellcontrolled experiments. 

E. Assays would be developed that not only evaluate the immunogenicity of 

relevant immunity in different systems, but that can also be used to predict those 

factors for screening of novel compounds for continuing development of 

immunointervention agents. Assays would concentrate on markers of relevant 

immunity and/or clinical endpoints and not on simple antigen quantitation. 

5. Proposed Collaborations: Industrial partners, Food and Drug Administration 

6. Anticipated project duration: 5 years+ 

7. Impact: 

A. Reduction in adverse reactions to vaccines. 

B. More optimal use of vaccines. 

2003 ARS Immunology Research Worskshop 99 
Immunity and the Pathogenesis of Diseases 

Working Group Leaders:

• Don Knowles, Research Leader, Animal Disease Research Unit, ARS, Pullman, WA 

• David Swayne, Research Leader, Southeast Poultry Research Laboratory, ARS, 

Athens, GA 

• Ray Waters, Veterinary Medical Officer, Bacterial Diseases of Livestock Research 

Unit, National Animal Disease Center, ARS, Ames, IA 

Research Priority 1

Characterization of immunity in the newborn 

1. Background: Immunointervention strategies for food producing animals are primarily 

targeted at neonatal/juvenile animals. Additionally, optimal development during the 

neonatal period is critical for attainment of full production of the animal. Immune 

development (ontogeny) and host/pathogen interactions of newborn livestock and 

poultry, however, are poorly characterized. 

2. Issue: Numerous developmental, maternal, environmental, and infectious factors 

influence immune responses elicited by neonates. Characterization of these influences on 

immune development would enhance strategies to improve immunity. 

3. Research Objectives: 

A. Determine the impact of maternal immunity on neonatal protection and immune 

response 

B. Determine the responses of neonates to vaccines 

C. Determine the impact of animal husbandry and environmental factors on neonatal 

immunity 

4. Proposed Accomplishments: 

A. Identified parameters of immune competence in the newborn 

B. Optimized vaccine strategies for level and duration of immunity 

C. Develop new tools for evaluation of immune competence 

5. Proposed Collaborations: Collaborations within ARS – across facilities, CRIS 

projects, and disciplines are in place as well as collaborations with industry and academic 

institutions. Additional collaborations should be added as needed. Certain aspects of the 

project will require international collaborations (e.g., Asia, United Kingdom, Australia, 

New Zealand, South Africa), of which many are already established. 

6. Anticipated project duration: Considering the breadth and impact of this project, it 

will likely be an on-going project. 

7. Impact: Improved strategies to enhance neonatal immunity will result and thereby 

improving production. More specifically, improved methods to enhance vaccine 

strategies targeting the neonate and maternal immunity as well as methods to limit factors 

2003 ARS Immunology Research Worskshop 100 
(e.g., environmental, husbandry, nutritional) that negatively impact neonatal immune 

development will result.

Research Priority 2

Determination of protective immunity to infectious diseases 

1. Background/Issue: An understanding of protective immunity is often required for 

development of successful strategies to limit disease. Unfortunately, specific protective 

mechanisms for most infectious diseases of food producing animals are not known. 

2. Issue: Our understanding of protective immunity and how to assess it is limited. As a 

result, this limits advances in development of immunity and immune strategies to control 

infectious diseases.

3. Research Objectives: 

A. Develop and validate challenge models for vaccine efficacy 

B. Determination of mechanisms of protection 

4. Proposed Accomplishments: 

A. Develop validated vaccine challenge models 

B. Identify mechanisms of immune protection 

5. Proposed Collaborations: Collaborations within ARS – across facilities, CRIS 

projects, and disciplines are in place as well as collaborations with industry and academic 

institutions. Certain aspects of the project will require international collaborations (e.g., 

Asia, United Kingdom, Australia, New Zealand, South Africa), of which many are 

already established. 

6. Anticipated project duration: In general, these are on-going projects with specific 

program funding. 

7. Impact: Establishment of infection model systems and correlates of protection will 

enhance our ability to develop effective immunointervention strategies. 

Research Priority 3

Regulation of the immune response in control of disease 

1. Background/Issue: Exacerbation or induction of disease often results from

inappropriate or excessive host responses. Understanding mechanisms of how immunity 

impacts disease development and resolution will improve our ability to control disease.

2. Issue: Some diseases result from the host immune response. Lack of understanding of 

this phenomenon for specific diseases has limited progress in prevention and treatment 

strategies. Important to this issue is the development of and/or sharing of reagents (such 

as monoclonal antibodies) which allow measurement and characterization of specific 

components of the immune response. 

3. Research Objectives: 

A. Determine the mechanisms of immunosuppression 

B. Determine the mechanisms of pathogen persistence 

C. Determine the role of autoimmunity in disease pathogenesis 

2003 ARS Immunology Research Worskshop 101 
4. Proposed Accomplishments: 

A. A determination of the role of pathogen persistence in morbidity and mortality 

B. An understanding of the role of immunosuppression in pathogenesis of disease 

C. New strategies to minimize autoimmunity in infectious disease following 

pathogen exposure 

5. Proposed Collaborations: Collaborations within ARS – across facilities, CRIS 

projects, and disciplines are in place as well as collaborations with industry and academic 

institutions. Additional collaborations should be added as needed. Certain aspects of the 

project will require international collaborations (e.g., Asia, United Kingdom, Australia, 

New Zealand, South Africa), of which many are already established. Develop a 

collaborative network for reagent development and exchange. 

6. Anticipated project duration: Due to this projects inherent complexity, the project 

duration is difficult to define. 

7. Impact: Improved methods to limit immune-associated disease and persistence of 

pathogens thereby increasing production and decrease spread of disease, respectively. 

2003 ARS Immunology Research Worskshop 102 
Nutrition and Immune Function Interactions 

Working Group Leaders: • Theodore Elsasser, Research Animal Scientist, Growth Biology Laboratory, Animal 

and Natural Resources Institute, ARS, Beltsville, MD 

• Simin Meydani, Chief, Nutritional Immunology Laboratory, Jean Mayer USDA Human 

Nutrition Research Center on Aging, ARS, Tufts University, Boston, MA 

• Joe Urban, Research Leader, Nutrient Requirements and Functions Laboratory, Beltsville 

Human Nutrition Research Center, ARS, 

 Beltsville, MD 

Research Priority 1

Defining nutrition related biomarkers that measure risk of disease. 

1. Background: Risk of disease during periods of physiological stress, such as birth, 

lactation, weaning, vaccination, ageing, metabolic disease, etc, can be ameliorated by 

appropriate nutritional support and intervention strategies. 

2. Issue: Nutritional biomarkers to assess optimal health, recommendations for nutritionally 

based interventions and monitoring of biomarkers to predict clinical outcome are needed. 

3. Research Objective: 

A. Evaluate markers of immune function and status that respond to nutritional 

modification. 

B. Identify dietary components that improve immune function and support optimal 

health. 

4. Proposed Accomplishments:

A. Develop reagents to identify immune markers for food production animals and 

man that respond to diet. 

B. Validate animal models to test nutrient and immune interactions in man. 

C. Quantitative risk/benefit evaluation of the dietary strategy to the appropriateness 

of the response. 

D. Identify specific nutrients with beneficial immune modulating properties that 

optimize immune function. 

5. Proposed Collaborations: Develop ARS Nutrition Network to share information on 

animal and human models of nutrient interaction with immune function. Need for webbased information sharing. 

6. Anticipated project duration: Significant progress should be made across species for 

identification/characterization of biomarkers within 5 years.

7. Impact: Sharing information on the variety of ARS-directed studies on animal models 

that measure the effect of nutrition on infectious disease and physiological stressors with 

human models of chronic inflammatory disease will provide a robust evaluation of multifunctional aspects of immune function and eliminate redundancy. This will improve 

2003 ARS Immunology Research Worskshop 103 
information exchange and identify novel markers that indicate the status of animal and 

human health related to diet. 

Research Priority 2

Identification of nutrient and gene interactions. 

1. Background: Novel dietary biomarkers and their associated genes could identify 

subpopulations of healthy or animals and humans at risk for disease. Genetic 

identification in animals of host resistance to important production and foodborne 

diseases and in humans of risk factors for chronic and genetic diseases is increasing. 

Collating these data will assist in designing dietary strategies that promote a healthy 

phenotype.

2. Issue: Identify and characterize genes that define disease risk factors in humans and 

determine if gene expression is responsive to dietary interventions. Evaluate the effect of 

nutrition on promoting optimal health in genetically defined disease resistant livestock. 

3. Research Objectives: 

A. Identify and characterize host genes that control important immunologic 

responses and determine responsiveness to diet. 

B. Define gene expression patterns and regulatory phenomena that are sensitive to 

diet and affect the level of host resistance 

C. Utilize genomic information from human and animal projects to reveal novel 

nutrient and immune interactions. 

D. Evaluate nutrients that activate immune gene transcription elements as modifiers 

of immune function. 

4. Proposed Accomplishments:

A. Mapping projects will identify pathogen and stress resistant livestock populations 

that can be targeted for nutritional studies related to health outcomes. 

B. Technical advances in gene expression/regulation approaches, includes 

microarray, real-time RT-PCR, proteomic studies, RNAi technology, etc. can be 

used to evaluate nutrient and gene interactions related to disease, inflammation 

and stress. 

C. Dietary interventions with genetically identified populations at risk for 

immunologically based diseases such as asthma and allergy, inflammatory bowel 

diseases, type-2 diabetes, obesity, etc. can be evaluated for efficacy. 

5. Proposed Collaborations: Work with scientists in NP101, NP106 and NP107 

programs to utilize genomics tools or identify animal models that can be used for 

nutritional studies. Partner with NIH and the human genome project to identify at risk 

populations for nutritional studies. 

6. Anticipated project duration: Significant progress should be made on gene 

identification, characterization and the role of nutrition in functional expression studies 

within 5 years. 

7. Impact: 

A Integrated strategies for control of pathogens and stress in livestock as well as 

inflammatory diseases in humans will include dietary recommendations. 

2003 ARS Immunology Research Worskshop 104 
B. The emergence of genetic profiles will include dietary recommendations for 

human life style and animal management to optimize health. 

Research Priority 3

Nutrient-dependent regulation of innate and acquired immunity. 

1. Background: Immunological stress affects nutrient partitioning and nutrient utilization 

affects immune system efficacy. Appropriate nutritional interventions will affect most 

components of innate and acquired immunity and influence disease susceptibility, 

vaccination, immunological memory, and the appropriateness and regulation of a 

balanced immune response. 

2. Issue: Maintenance of optimal health and its restoration following stress from

infection or inflammation are dependent on adequate nutrition. Improved understanding 

of the interaction between innate and acquired immunity and the mechanisms of immune 

regulation will provide an opportunity to examine specific nutrient requirements for these 

responses. 

3. Research Objectives: 

A. Define the affects of diet on stress and age-related changes in innate and acquired 

immunity. 

B. Define the effect of diet on the ontogeny of the immune system and interventions 

that provide a balanced response to stress and disease. 

C. Evaluate the role of immunity in specific nutrient uptake from the intestine.

D. Define specific nutrient requirements for effective and appropriate vaccination 

and immunological memory, and for amelioration of periods of immune 

deficiency such as during pregnancy, parturition, pediatric development, ageing, 

stress, etc. 

F. Evaluate the role of nutrition in the amelioration of inflammation and immune 

pathogenesis. 

4. Proposed Accomplishments: 

A. Knowledge of nutrient requirements for the development and regulation of 

immunity in mothers, neonates and the elderly. 

B. Shared discoveries on the role of specific nutrients in cell and tissue function 

including receptor mobility, cell signaling pathways, regulation of transcription, 

mucosal barrier function and physiology, trace mineral-dependent enzymatic 

activity, reactive oxygen and nitrogen species, lipid inflammatory mediators, 

carbohydrate glycosyltransferases, oral re hydration solutions, neural transmitters, 

etc. 

C. More effective vaccines and bioprophylactics. 

D. Nutritional recommendations to improve immune efficacy during periods of stress 

and disease.

5. Proposed Collaborations: Partnerships with food industry and animal health 

companies; ARS plant scientists to provide modified and functional food sources; 

nutritionists involved with nutrient bioavailability. 

2003 ARS Immunology Research Worskshop 105 
6. Anticipated project duration: Significant progress in immune regulation and nutrient 

interactions are assured given the high level of interest. 

7. Impact: 

A. Provide recommendations for specific nutrients that contribute to reduced stress 

and enhanced disease resistance. 

B. Identification of nutrients required for improved vaccine efficacy. 

C. Provide identification of diets that enhance immune development and reduce 

inflammation. 

Research Priority 4

Nutrition and applied immunology 

1. Background: Food ingestion and the nutrients provided affect resident microflora in 

the oral, urogenital and gastrointestinal track, and interact with neural and endocrine 

pathways to reduce stress. Functional nutrient deficiencies or excess affect the balance of 

the immune system leading to inflammation or inappropriate responses to infection or 

stress. 

2. Issue: Extend the matrix of information to include indirect interactions between 

nutrients and the immune system as well as the role of nutrients as modulators of immune 

function.

3. Research Objectives: 

A. Define probiotic microbial populations that affect immune development and 

functional resistance at mucosal surfaces and on the efficacy of systemic 

immunity, and the diets (prebiotics) that support appropriate resident or ingested 

microbial populations. 

B. Establish a repository of microbial strains that benefit host species and apply 

genetic or selection tools to improve their application. 

C. Study the role of specific nutrients as modulators of immune function, regulators 

of inflammation, and as adjuvants. 

D. Evaluate the interaction between diet, environment, behavior and stress on the 

efficacy of immune function. 

4. Proposed Accomplishments:

A. Identification of functional dietary deficiencies or super nutritional intake of 

nutrients that result from behavior, environment or stress that affect optimal 

immune function. 

B. Establishment of animal models that evaluate the long term consequences of diet 

on immune function and chronic disease. 

C. Definition of the affect of inflammation and immune pathogenesis (obesity, 

chronic disease, infection, stress) on nutrient uptake and partitioning. 

2003 ARS Immunology Research Worskshop 106 
C. Application of genomics information to characterize microbial genes that 

improve host health by the production of useful metabolics, immune stimulators, 

and competition for sites of pathogen invasion. 

5. Proposed Collaborations: Interactions with food industry and animal health 

companies; food survey groups; microbiologists; food technologists; EPA; FDA. 

6. Anticipated project duration: Significant progress should be visible within 5 years

with support of intra structure and cooperation. 

7. Impact: 

A. Definition of patentable food products that affect human and animal immune 

function. 

B. Better understanding of the interaction between dietary choices and environment 

on immune function and optimal health. 

2003 ARS Immunology Research Worskshop 107 
Bibliography of ARS Immunology Research 2000-2003 

This bibliography contains selected citations of immunology research being performed at 

ARS Research Centers throughout the United States. The citations were drawn from the 

CRIS database maintained by the USDA, Cooperative State Research, Education, and 

Extension Service (http://cris.csrees.usda.gov/), AgricOLA database at the National 

Agricultural Library, CAB Abstracts in the United Kingdom, and MedLine at the 

National Library of Medicine. It is provided to give an overview of the many facets of 

ARS immunology research. 

It is organized alphabetically by State; within each State it is organized alphabetically by 

the last name of the staff listed. 

2003 ARS Immunology Research Worskshop 108 
Auburn, Alabama 

Aquatic Animal Health Research 

http://www.ars.usda.gov/pandp/locations/locations.htm?modecode=64-20-15-00

Evans JJ

Evans, J.J., Shoemaker, C.A., Klesius, P. H. Distribution of Streptococcus iniae 

in hybrid striped bass (Morone chrysops X Morone saxatilis) following nare 

inoculation. Aquaculture. 2001. v. 194. p. 233-243. 

Klesius, P.H., Shoemaker, C.A., Evans, J.J. Vaccination: A health management 

practice for preventing diseases caused by Streptococcus in tilapia and other 

cultured fish. Fitzsimmons, K., Filho, Jomar Carvalho, editors. Panorama da 

AQUICULTURA, Brazil. Tilapia Aquaculture in the 21st Century Proceedings 

from the Fifth International Symposium on Tilapia Aquaculture. 2000. p. 558- 

564. 

Klesius, P.H., Shoemaker, C.A., Evans, J.J. Streptococcus iniae vaccine. Patent 

allowance given February 28, 2001. Application No. 09/513,143. 

Klesius, P.H., Evans, J.J., Shoemaker, C.A. Communicable bacterial diseases in 

fish farming: Prevention by vaccination and immunostimulation. Obaldia III, N., 

editor. Panama. Proceedings of the XVII Panamerican Congress of Veterinary 

Sciences. 2000. p. 13-14. 

Shelby-RA; Klesius-PH; Shoemaker-CA; Evans-JJ Passive immunization of 

tilapia, Oreochromis niloticus (L.), with anti-Streptococcus iniae whole sera. 

Journal-of-Fish-Diseases. 2002, 25: 1, 1-6. 

Shelby,-R.A.; Shoemaker,-C.A.; Evans,-J.J.; Klesius,-P.H. Development of an 

indirect ELISA to detect humoral response to Streptococcus iniae infection of 

Nile tilapia, Oreochromis niloticus. J-appl-aquac. Binghamton, NY : Food 

Products Press, 1991-. 2001. v. 11 (3) p. 35-44. 

Shoemaker-CA; Klesius-PH; Evans-JJ. In ovo methods for utilizing the modified 

live Edwardsiella ictaluri vaccine against enteric septicemia in channel catfish. 

Aquaculture. 2002, 203: 3-4, 221-227. 

Shoemaker, C.A., Klesius, P.H., Evans, J.J. Prevalence of Streptococcus iniae in 

tilapia, hybrid striped bass and channel catfish from fish farms in the US. 

American Journal of Veterinary Research. 2001. v. 62(2). p. 174-177. 

Shoemaker, C.A., Klesius, P.H., Evans, J.J. Diseases of tilapia with emphasis on 

economically important pathogens. Fitzsimmons, K., Filho, Jomar Carvalho, 

2003 ARS Immunology Research Worskshop 109 
editors. Panorama da AQUICULTURA, Brazil. Tilapia Aquaculture in the 21st 

Century Proceedings from the Fifth International Symposium on Tilapia 

Aquaculture. 2000. p. 565-572. 

Xu, D.H., Klesius, P.H., Shoemaker, C.A., Evans, J.J. The early development of 

Ichthyopthirius multifiliis in channel catfish tissue in vitro. Journal of Aquatic 

Animal Health. 2000. v. 12. p. 290-296. 

Klesius PH

Barros, M.M.; Lim,-C.; Klesius,-P.H. Effect of soybean meal replacement by 

cottonseed meal and iron supplementation on growth, immune response and 

resistance of Channel Catfish (Ictalurus puctatus) to Edwardsiella ictaluri 

challenge.Aquaculture. Amsterdam : Elsevier Pub. Co., c1972-. May 24, 2002. v. 

207 (3/4) p. 263-279. 

Evans, J.J., Shoemaker, C.A., Klesius, P. H. Distribution of Streptococcus iniae 

in hybrid striped bass (Morone chrysops X Morone saxatilis) following nare 

inoculation. Aquaculture. 2001. v. 194. p. 233-243. 

Klesius, P.H., Shoemaker, C.A., Evans, J.J. Streptococcus iniae vaccine. Patent 

allowance given February 28, 2001. Application No. 09/513,143. 

Klesius, P.H., Shoemaker, C.A., Evans, J.J. and Lim, C. Vaccines: Prevention of 

diseases in aquatic animals. Lim, C., Webster, C., editors. Haworth Press, Inc., 

New York, NY. Nutrition and Fish Health. 2001. p. 317-335. 

Lim-C; Klesius-PH Influence of dietary levels of magnesium on growth, tissue 

mineral content, and resistance of channel catfish Ictalurus punctatus challenged 

with Edwardsiella ictaluri. Journal-of-the-World-Aquaculture-Society. 2003, 34: 

1, 18-28. 

Lim, C., Shoemaker, C.A., Klesius, P.H. The effect of ascorbic acid on immune 

response in fish. Dabrowski, K., editor. CRC Press, Boca Raton, FL. Ascorbic 

Acid in Aquatic Organisms. 2001. p. 149-166. 

Lim, C., Klesius, P.H., Webster, C.D. The role of dietary phosphorus, zinc and 

selenium in fish health. Lim, C., Webster, C., editors. Haworth Press, Inc., New 

York, NY. Nutrition and Fish Health. 2001. p. 201-212. 

Lim, C., Klesius, P.H., Shoemaker, C.A. Dietary iron and fish health. Lim, C., 

Webster, C., editors. Haworth Press, Inc., New York, NY. Nutrition and Fish 

Health. 2001. p. 189-199. 

2003 ARS Immunology Research Worskshop 110 
Shelby-RA; Klesius-PH; Shoemaker-CA; Evans-JJ Passive immunization of 

tilapia, Oreochromis niloticus (L.), with anti-Streptococcus iniae whole sera. 

Journal-of-Fish-Diseases. 2002, 25: 1, 1-6. 

Shelby,-R-A; Shoemaker,-C-A; Klesius,-P-H Development of an immunoassay 

to measure the humoral immune response of hybrid striped bass Morone 

chrysopsxM. saxatilis. Veterinary-immunology-and-immunopathology 2003 Feb 

10; 91(3-4): 217-25. 

Shelby,-R.A.; Shoemaker,-C.A.; Klesius,-P.H. Detection of humoral response to 

Streptococcus iniae infection of Nile tilapia, Oreochromis niloticus, by a 

monoclonal antibody-based ELISA. Journal of Appl Aquaculture Binghamton, 

NY : Food Products Press, 1991-. 2002. v. 12 (3) p. 23-31. 

Shelby,-R.A.; Shoemaker,-C.A.; Evans,-J.J.; Klesius,-P.H. Development of an 

indirect ELISA to detect humoral response to Streptococcus iniae infection of 

Nile tilapia, Oreochromis niloticus. J-appl-aquac. Binghamton, NY : Food 

Products Press, 1991-. 2001. v. 11 (3) p. 35-44. 

Shoemaker, C.A., Klesius, P.H., Lim, C. Immunity and disease resistance in fish. 

Lim, C., Webster, C., editors. Haworth Press, Inc., New York, NY. Nutrition and 

Fish Health. 2001. p. 149-162. 

Shoemaker-CA; Klesius-PH; Evans-JJ. In ovo methods for utilizing the modified 

live Edwardsiella ictaluri vaccine against enteric septicemia in channel catfish. 

Aquaculture. 2002, 203: 3-4, 221-227. 

Shoemaker, C.A., Klesius, P.H., Evans, J.J. Prevalence of Streptococcus iniae in 

tilapia, hybrid striped bass and channel catfish from fish farms in the US. 

American Journal of Veterinary Research. 2001. v. 62(2). p. 174-177. 

Xu,-D-H; Klesius,-P-H. Protective effect of cutaneous antibody produced by 

channel catfish, Ictalurus punctatus (Rafinesque), immune to Ichthyophthirius 

multifiliis Fouquet on cohabited non-immune catfish. J-Fish-Dis. 2003 May; 

26(5): 287-91. 

Xu, D.H., Klesius, P.H., Shoemaker, C.A. Effect of lectins on the invasion of 

Ichthyophthirius to channel catfish tissues. Diseases of Aquatic Organisms. 2001. 

v. 45(2). p. 115-120. 

Yildirim-M; Lim-C; Wan-PJ; Klesius-PH Growth performance and immune 

response of channel catfish (Ictalurus puctatus) fed diets containing graded levels 

of gossypol-acetic acid.Aquaculture. 2003, 219: 1-4, 751-768. 

2003 ARS Immunology Research Worskshop 111 
Lim CE

Barros, M.M.; Lim,-C.; Klesius, P.H. Effect of soybean meal replacement by 

cottonseed meal and iron supplementation on growth, immune response and 

resistance of Channel Catfish (Ictalurus puctatus) to Edwardsiella ictaluri 

challenge.Aquaculture. Amsterdam : Elsevier Pub. Co., c1972-. May 24, 2002. v. 

207 (3/4) p. 263-279. 

Klesius, P.H., Shoemaker, C.A.; Evans, J.J.; and Lim, C. Vaccines: Prevention of 

diseases in aquatic animals. Lim, C., Webster, C., editors. Haworth Press, Inc., 

New York, NY. Nutrition and Fish Health. 2001. p. 317-335. 

Lim, C.; Klesius, P.H. Influence of dietary levels of magnesium on growth, tissue 

mineral content, and resistance of channel catfish Ictalurus punctatus challenged 

with Edwardsiella ictaluri. Journal-of-the-World-Aquaculture-Society. 2003, 34: 

1, 18-28. 

Lim, C.; Klesius, P.H.; Shoemaker, C.A. Dietary iron and fish health. Lim, C., 

Webster, C., editors. Haworth Press, Inc., New York, NY. Nutrition and Fish 

Health. 2001. p. 189-199. 

Lim, C.; Webster, C.D.; editors. Nutrition and Fish Health. Haworth Press, Inc., 

New York, NY. 2001. 365 pp. 

Lim, C.; Klesius, P.H.; Webster, C.D. The role of dietary phosphorus, zinc and 

selenium in fish health. Lim, C.; Webster, C.; editors. Haworth Press, Inc., New 

York, NY. Nutrition and Fish Health. 2001. p. 201-212. 

Lim, C.; Shoemaker, C.A.; Klesius, P.H. The effect of ascorbic acid on immune 

response in fish. Dabrowski, K., editor. CRC Press, Boca Raton, FL. Ascorbic 

Acid in Aquatic Organisms. 2001. p. 149-166. 

Shoemaker, C.A.; Klesius, P.H.; Lim, C. Immunity and disease resistance in fish. 

Lim, C.; Webster, C.; editors. Haworth Press, Inc., New York, NY. Nutrition and 

Fish Health. 2001. p. 149-162. 

Yildirim, M.; Lim, C.; Wan, P.J.; Klesius,P.H. Growth performance and immune 

response of channel catfish (Ictalurus puctatus) fed diets containing graded levels 

of gossypol-acetic acid.Aquaculture. 2003, 219: 1-4, 751-768. 

Shelby RA

Shelby, R.A.; Klesius, P.H.; Shoemaker, C.A.; Evans, J.J. Passive immunization 

of tilapia, Oreochromis niloticus (L.), with anti-Streptococcus iniae whole sera. 

Journal-of-Fish-Diseases. 2002, 25: 1, 1-6. 

2003 ARS Immunology Research Worskshop 112 
Shelby, R.A.; Shoemaker, C.A.; Klesius, P.H. Development of an immunoassay 

to measure the humoral immune response of hybrid striped bass Morone 

chrysopsxM. saxatilis. Veterinary-immunology-and-immunopathology 2003 Feb 

10; 91(3-4): 217-25. 

Shelby, R.A.; Shoemaker, C.A.; Klesius, P.H. Detection of humoral response to 

Streptococcus iniae infection of Nile tilapia, Oreochromis niloticus, by a 

monoclonal antibody-based ELISA. Journal of Appl Aquaculture Binghamton, 

NY : Food Products Press, 1991-. 2002. v. 12 (3) p. 23-31. 

Shelby, R.A.; Shoemaker, C.A.; Evans, J.J.; Klesius, P.H. Development of an 

indirect ELISA to detect humoral response to Streptococcus iniae infection of 

Nile tilapia, Oreochromis niloticus. J-appl-aquac. Binghamton, NY : Food 

Products Press, 1991-. 2001. v. 11 (3) p. 35-44. 

Shoemaker CA

Evans, J.J.; Shoemaker, C.A.; Klesius, P. H. Distribution of Streptococcus iniae 

in hybrid striped bass (Morone chrysops X Morone saxatilis) following nare 

inoculation. Aquaculture. 2001. v. 194. p. 233-243. 

Klesius, P.H.; Shoemaker, C.A.; Evans, J.J. Streptococcus iniae vaccine. Patent 

allowance given February 28, 2001. Application No. 09/513,143. 

Klesius, P.H.; Shoemaker, C.A.; Evans, J.J. Vaccination: A health management 

practice for preventing diseases caused by Streptococcus in tilapia and other 

cultured fish. Fitzsimmons, K., Filho, Jomar Carvalho, editors. Panorama da 

AQUICULTURA, Brazil. Tilapia Aquaculture in the 21st Century Proceedings 

from the Fifth International Symposium on Tilapia Aquaculture. 2000. p. 558-

564. 

Klesius, P.H.; Evans, J.J.; Shoemaker, C.A. Communicable bacterial diseases in 

fish farming: Prevention by vaccination and immunostimulation. Obaldia III, N., 

editor. Panama. Proceedings of the XVII Panamerican Congress of Veterinary 

Sciences. 2000. p. 13-14. 

Klesius, P.H.; Shoemaker, C.A.; Evans, J.J.; and Lim, C. Vaccines: Prevention 

of diseases in aquatic animals. Lim, C., Webster, C., editors. Haworth Press, Inc., 

New York, NY. Nutrition and Fish Health. 2001. p. 317-335. 

Lim, C.; Klesius, P.H.; Shoemaker, C.A. Dietary iron and fish health. Lim, C., 

Webster, C., editors. Haworth Press, Inc., New York, NY. Nutrition and Fish 

Health. 2001. p. 189-199. 

2003 ARS Immunology Research Worskshop 113 
Lim, C.; Shoemaker, C.A.; Klesius, P.H. The effect of ascorbic acid on immune 

response in fish. Dabrowski, K., editor. CRC Press, Boca Raton, FL. Ascorbic 

Acid in Aquatic Organisms. 2001. p. 149-166. 

Shelby, R.A.; Klesius, P.H.; Shoemaker, C.A.; Evans, J.J. Passive immunization 

of tilapia, Oreochromis niloticus (L.), with anti-Streptococcus iniae whole sera. 

Journal-of-Fish-Diseases. 2002, 25: 1, 1-6. 

Shelby, R.A.; Shoemaker, C.A.; Evans, J.J.; Klesius, P.H. Development of an 

indirect ELISA to detect humoral response to Streptococcus iniae infection of 

Nile tilapia, Oreochromis niloticus. J-appl-aquac. Binghamton, NY : Food 

Products Press, 1991-. 2001. v. 11 (3) p. 35-44. 

Shelby, R.A.; Shoemaker, C.A.; Klesius, P.H. Development of an immunoassay 

to measure the humoral immune response of hybrid striped bass Morone chrysops 

x M. saxatilis. Veterinary-Immunology-and-Immunopathology. 2003, 91: 3-4, 

217-225. 

Shelby, R.A.; Shoemaker, C.A.; Klesius, P.H. Detection of humoral response to 

Streptococcus iniae infection of Nile tilapia, Oreochromis niloticus, by a 

monoclonal antibody-based ELISA. Journal of Applied Aquaculture 2002. v. 12 

(3) p. 23-31. 

Shoemaker, C.A.; Klesius, P.H.; Evans, J.J. In ovo methods for utilizing the 

modified live Edwardsiella ictaluri vaccine against enteric septicemia in channel 

catfish. Aquaculture. 2002, 203: 3-4, 221-227. 

Shoemaker, C.A.; Klesius, P.H.; Evans, J.J. Prevalence of Streptococcus iniae in 

tilapia, hybrid striped bass and channel catfish from fish farms in the US. 

American Journal of Veterinary Research. 2001. v. 62(2). p. 174-177. 

Shoemaker, C.A.; Klesius, P.H.; Evans, J.J. Diseases of tilapia with emphasis on 

economically important pathogens. Fitzsimmons, K., Filho, Jomar Carvalho, 

editors. Panorama da AQUICULTURA, Brazil. Tilapia Aquaculture in the 21st 

Century Proceedings from the Fifth International Symposium on Tilapia 

Aquaculture. 2000. p. 565-572. 

Shoemaker, C.A.; Klesius, P.H.; Lim, C. Immunity and disease resistance in fish.

Lim, C., Webster, C., editors. Haworth Press, Inc., New York, NY. Nutrition and 

Fish Health. 2001. p. 149-162. 

Xu, D.H.; Klesius, P.H.; Shoemaker, C.A.; Evans, J.J. The early development of 

Ichthyopthirius multifiliis in channel catfish tissue in vitro. Journal of Aquatic 

Animal Health. 2000. v. 12. p. 290-296. 

2003 ARS Immunology Research Worskshop 114 
Xu, D.H.; Klesius, P.H.; Shoemaker, C.A. Effect of lectins on the invasion of 

Ichthyophthirius to channel catfish tissues. Diseases of Aquatic Organisms. 2001. 

v. 45(2). p. 115-120. 

Xu D

Xu, D.H.; Klesius, P.H.; Protective effect of cutaneous antibody produced by 

channel catfish, Ictalurus punctatus (Rafinesque), immune to Ichthyophthirius 

multifiliis Fouquet on cohabited non-immune catfish. J-Fish-Dis. 2003 May; 

26(5): 287-91. 

Xu, D.H.; Klesius, P.H.; Shoemaker, C.A. Effect of lectins on the invasion of 

Ichthyophthirius to channel catfish tissues. Diseases of Aquatic Organisms. 

2001.v. 45(2). p. 115-120. 

Xu, D.H.; Klesius, P.H.; Shoemaker, C.A.; Evans, J.J. The early development of 

Ichthyopthirius multifiliis in channel catfish tissue in vitro. Journal of Aquatic 

Animal Health. 2000. v. 12. p. 290-296. 

Fayetteville, Arkansas 

Poultry Production and Products Safety Research 

http://www.ars.usda.gov/pandp/locations/locations.htm?modecode=62-26-05-00

Balog JM

Huff, G.R.; Huff, W.E.; Balog, J.M.; Rath, N.C. The effects of behavior and 

environmental enrichment on disease resistance of turkeys. Brain, Behavior, and 

Immunity 2003 Oct; 17(5): 339-49. 

Huff, G.R.; Huff, W.E.; Balog, J.M.; Rath, N.C.; Xie, H.; Horst, R.L. Effect of 

dietary supplementation with vitamin D metabolites in an experimental model of 

turkey osteomyelitis complex. Poultry Science July 2002. v. 81 (7) p. 958-965. 

Huff, G.R.; Huff, W.E.; Rath, N.C.; Balog, J.M. Turkey osteomyelitis complex. 

Poultry Science July 2000. v. 79 (7) p. 1050-1056. 

Xie, H.; Huff, G.R.; Huff, W.E.; Balog, J.M.; Holt,P.; Rath, N.C. Identification of

ovotransferrin as an acute phase protein in chickens. Poultry Science 2002 Jan; 

81(1): 112-20. 

2003 ARS Immunology Research Worskshop 115 
Donoghue AM

Huff, W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Xie, H.; Moore, P.A. Jr.; 

Donoghue, A.M. Prevention of Escherichia coli respiratory infection in broiler 

chickens with bacteriophage (SPR02). Poultry-science Apr 2002. v. 81 (4) p. 437-

441. 

Huff, W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Xie, H.; Moore, P.A., Jr.; 

Donoghue, A.M. Efficacy of bacteriophage to prevent Escherichia coli 

respiratory infection in broiler chickens when administered in the drinking water 

prior to challenge. Poultry Science. 2001. v. 80(Suppl.1): Abstract p. 238. 

Huff W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Xie, H.; Moore, P.A.; Jr., 

Donoghue, A.M. Bacteriophage prevents Escherichia coli respiratory infection in 

broiler chickens. Poultry Science. 2001. v. 80(Suppl. 1): Abstract p. 237. 

Huff GR

Huff, G.R.; Huff, W.E.; Balog, J.M.; Rath, N.C. The effects of behavior and 

environmental enrichment on disease resistance of turkeys. Brain, Behavior, and 

Immunity 2003 Oct; 17(5): 339-49. 

Huff, G.R.; Huff, W.E.; Balog, J.M.; Rath, N.C. Effect of early handling of 

turkey poults on later responses to a dexamethasone Escherichia coli challenge. 1. 

Production values and physiological response. Poultry Science. 2001. v. 80. p. 

1305-1313. 

Huff, G.R.; Huff, W.E.; Balog, J.M.; Rath, N.C. Effect of early handling of 

turkey poults on later responses to a dexamethasone Escherichia coli challenge. 2. 

Resistance to air sacculitis and turkey osteomyelitis complex. Poultry Science. 

2001. v. 80. p. 1314-1322. 

Huff, G.R.; Huff, W.E.; Rath, N.C.; Balog, J.M. Effect of feed supplementation 

with vitamin D metabolites in a dexamethasone-Escherichia coli challenge model 

of turkey osteomyelitis complex. Poultry Science. 2001. v. 80(Suppl. 1): Abstract 

p. 238. 

Huff, W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Donoghue, A.M. Prevention of 

Escherichia coli infection in broiler chickens with a bacteriophage aerosol spray. 

Poultry-Science. 2002, 81: 10, 1486-1491. 

Huff, W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Xie, H.; Moore, P.A. Jr.; 

Donoghue, A.M. Prevention of Escherichia coli respiratory infection in broiler 

chickens with bacteriophage (SPR02). Poultry-science Apr 2002. v. 81 (4) p. 437-

441. 

2003 ARS Immunology Research Worskshop 116 
Huff, W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Xie, H.; Moore, P.A. Jr.; 

Donoghue, A.M. Bacteriophage prevents Escherichia coli respiratory infection in 

broiler chickens. Poultry Science. 2001. v. 80(Suppl. 1): Abstract p. 237. 

Huff, W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Xie, H.; Moore, P.A. Jr.; 

Donoghue, A.M. Efficacy of bacteriophage to prevent Escherichia coli respiratory 

infection in broiler chickens when administered in the drinking water prior to 

challenge. Poultry Science. 2001. v. 80(Suppl.1): Abstract p. 238. 

Xie, H.; Huff, G.R.; Huff, W.E.; Balog, J.M.; Rath, N.C. Effects of ovotransferrin 

on chicken macrophages and heterophil-granulocytes. Developmental and 

Comparative Immunology. 2002, 26: 9, 805-815. 

Xie, H.; Rath, N.C.; Huff, G.R.; Balog, J.M.; Huff, W.E. Inflammation-induced 

changes in serum modulate chicken macrophage function. Vet-immunolimmunopathol. Amsterdam : Elsevier. Aug 10, 2001. v. 80 (3/4) p. 225-235. 

Huff WE

Huff, G.R.; Huff, W.E.; Rath, N.C.; Balog, J.M. Effect of feed supplementation 

with vitamin D metabolites in a dexamethasone-Escherichia coli challenge model 

of turkey osteomyelitis complex. Poultry Science. 2001. v. 80(Suppl. 1): Abstract 

p. 238. 

Huff, G.R.; Huff, W.E.; Balog, J.M.; Rath, N.C. Effect of early handling of 

turkey poults on later responses to a dexamethasone Escherichia coli challenge. 1. 

Production values and physiological response. Poultry Science. 2001. v. 80. p. 

1305-1313. 

Huff, G.R.; Huff, W.E.; Balog, J.M.; Rath, N.C. Effect of early handling of 

turkey poults on later responses to a dexamethasone Escherichia coli challenge. 2. 

Resistance to air sacculitis and turkey osteomyelitis complex. Poultry Science. 

2001. v. 80. p. 1314-1322. 

Huff, W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Donoghue, A.M. Evaluation of 

aerosol spray and intramuscular injection of bacteriophage to treat an Escherichia 

coli respiratory infection. Poultry Science 2003 Jul; 82(7): 1108-12. 

Huff, W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Donoghue, A.M. Prevention of 

Escherichia coli infection in broiler chickens with a bacteriophage aerosol spray. 

Poultry-Science. 2002, 81: 10, 1486-1491. 

Huff, W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Xie, H.; Moore, P.A. Jr.; 

Donoghue, A.M. Prevention of Escherichia coli respiratory infection in broiler 

chickens with bacteriophage (SPR02). Poultry-science Apr 2002. v. 81 (4) p. 437-

441. 

2003 ARS Immunology Research Worskshop 117 
Huff, W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Xie, H.; Moore, P.A., Jr.; 

Donoghue, A.M. Bacteriophage prevents Escherichia coli respiratory infection in 

broiler chickens. Poultry Science. 2001. v. 80(Suppl. 1): Abstract p. 237. 

Huff, W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Xie, H.; Moore, P.A., Jr.; 

Donoghue, A.M. Efficacy of bacteriophage to prevent Escherichia coli respiratory 

infection in broiler chickens when administered in the drinking water prior to 

challenge. Poultry Science. 2001. v. 80(Suppl.1): Abstract p. 238. 

Xie, H; Huff, G.R.; Huff, W.E.; Balog, J.M.; Rath, N.C. Effects of ovotransferrin 

on chicken macrophages and heterophil-granulocytes. Developmental and 

Comparative Immunology. 2002, 26: 9, 805-815. 

Xie, H.; Rath, N.C.; Huff, G.R.; Balog, J.M.; Huff, W.E. Inflammation-induced 

changes in serum modulate chicken macrophage function. Vet-immunolimmunopathol. Amsterdam : Elsevier. Aug 10, 2001. v. 80 (3/4) p. 225-235. 

Rath NC

Huff, G.R.; Huff, W.E.; Rath, N.C.; Balog, J.M. Effect of feed supplementation 

with vitamin D metabolites in a dexamethasone-Escherichia coli challenge model 

of turkey osteomyelitis complex. Poultry Science. 2001. v. 80(Suppl. 1): Abstract 

p. 238. 

Huff, G.R.; Huff, W.E.; Balog, J.M.; Rath, N.C. Effect of early handling of 

turkey poults on later responses to a dexamethasone Escherichia coli challenge. 1. 

Production values and physiological response. Poultry Science. 2001. v. 80. p. 

1305-1313. 

Huff, G.R.; Huff, W.E.; Balog, J.M.; Rath, N.C. Effect of early handling of 

turkey poults on later responses to a dexamethasone Escherichia coli challenge. 2. 

Resistance to air sacculitis and turkey osteomyelitis complex. Poultry Science. 

2001. v. 80. p. 1314-1322. 

Huff, G.R.; Zheng, Q.; Newberry, L.A.; Huff, W.E.; Balog, J.M.; Rath, N.C.; 

Kim, K.S.; Martin, E.M.; Goeke, S.C.; Skeeles, J.K. Viral and bacterial agents 

associated with experimental transmission of infectious proventriculitis of broiler 

chickens. Avian Diseases Oct/Dec 2001. v. 45 (4) p. 828-843. 

Huff, W.E.; Huff, G.R.; Rath, N.C; Balog, J.M.; Donoghue, A.M. Prevention of 

Escherichia coli infection in broiler chickens with a bacteriophage aerosol spray. 

Poultry-Science. 2002, 81: 10, 1486-1491. 

Huff, W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Xie, H.; Moore, P.A. Jr.; 

Donoghue, A.M. Prevention of Escherichia coli respiratory infection in broiler 

2003 ARS Immunology Research Worskshop 118 
chickens with bacteriophage (SPR02). Poultry-science Apr 2002. v. 81 (4) p. 437-

441. 

Huff, W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Xie, H.; Moore, P.A., Jr.; 

Donoghue, A.M. Bacteriophage prevents Escherichia coli respiratory infection in 

broiler chickens. Poultry Science. 2001. v. 80(Suppl. 1): Abstract p. 237. 

Huff, W.E.; Huff, G.R.; Rath, N.C.; Balog, J.M.; Xie, H.; Moore, P.A. Jr.; 

Donoghue, A.M. Efficacy of bacteriophage to prevent Escherichia coli respiratory 

infection in broiler chickens when administered in the drinking water prior to 

challenge. Poultry Science. 2001. v. 80(Suppl.1): Abstract p. 238. 

Xie, H; Huff, G.R.; Huff, W.E.; Balog, J.M.; Rath, N.C. Effects of ovotransferrin 

on chicken macrophages and heterophil-granulocytes. Developmental and 

Comparative Immunology. 2002, 26: 9, 805-815. 

Xie, H.; Rath, N.C.; Huff, G.R.; Balog, J.M.; Huff, W.E. Inflammation-induced 

changes in serum modulate chicken macrophage function. Vet-immunolimmunopathol. Amsterdam : Elsevier. Aug 10, 2001. v. 80 (3/4) p. 225-235. 

Little Rock, Arkansas 

Arkansas Children's Nutrition Center 

http://www.ars.usda.gov/pandp/locations/locations.htm?modecode=62-51-10-00

Badger TM

Badger, T.M.; Hakkak, R.; Ronis, M.J.J. Developmental effects and health 

aspects of soy protein isolate, casein and whey in male and female rats. 

International Journal of Toxicology. 2001. v. 20. p. 165-175. 

Hakkak, R.; Korourian, S.; Fletcher, T.; Hale, K.; Ferguson, M.; Holder, D.; Ronis, 

M.J.J.; Rowlands, J.C.; Johnston, J.; Badger, T.M. Effects of whey protein 

consumption in different stages of life on protection of DMBA-induced mammary 

tumors in rats. Proceedings of the American Association of Cancer Research. 2001. 

v. 42. Abstract p. 311. 

Hakkak, R.; Korourian, S.; Irby, D.; Sweeny, J.; Weatherford, C.; Ronis, M.J.J.;

Rowlands, J.C.; Badger, T.M. Effects of whey protein consumption on initiation and 

promotion of DMBA-induced mammary tumors in rats. Proceedings of American 

Association for Cancer Research. 2001. v. 42. Abstract p. 892. 

Hakkak, R.; Korourian, S.; Ronis, M.J.J.; Johnston, J.; Badger, T.M. Lifetime soy 

protein isolate consumption protects against azoxymethane- induced colon tumors in

male rats. Cancer Letters. 2001. v. 166. p. 27-32. 

2003 ARS Immunology Research Worskshop 119 
Ronis, M.J.J.; Mercado, C.; Holder, D.; Sharp S.; Irby, D.; Hakkak, R.; Badger,

T.M. Inducer-specific effects of dietary protein source on CYP2B1 expression and 

activity in male rat liver. The FASEB Journal. 2001. v. 15. Abstract p. A627. 

Hakkak R

Badger, T.M.; Hakkak, R.; Ronis, M.J.J. Developmental effects and health aspects 

of soy protein isolate, casein and whey in male and female rats. International Journal 

of Toxicology. 2001. v. 20. p. 165-175. 

Hakkak, R.; Korourian, S.; Ronis, M.J.J.; Johnston J.; Badger, T.M. Lifetime soy 

protein isolate consumption protects against azoxymethane- induced colon tumors in

male rats. Cancer Letters. 2001. v. 166. p. 27-32. 

Prior RL

Han, S.N.; Meydani, M.; Wu, D.; Bender, B.S.; Smith, D.E.; Vina, J.; Cao, G.; 

Prior, R.L.; Meydani, S.N. Effect of long-term dietary antioxidant 

supplementation on influenza virus infection. Journals of Gerontology Biological 

Sciences and Medical Sciences. 2000. v. 55A. p. B1-B8. 

Davis, California 

Western Human Nutrition Research Center 

http://www.whnrc.usda.gov/

Hwang DH

Lee, J. Y.; Ye, J.; Gao, Z.; Youn, H. S.; Lee, W. H.; Zhao, L.; Hwang, D H.

(2003). Reciprocal modulation of TLR4signaling pathways involving MyD88 and 

PI3K/AKT by saturated and polyunsaturated fatty acids (J. Biol. Chem., in Press). 

Lee, J. Y.; Plakidas, A.; Lee, W.H.; Chanmugam, P.; Heikinnen, A.; Bray, G.A.; 

Hwang, D.H. Differentiall modulation of Toll-like receptors by fatty acids—

Preferential inhibition by n-3 polyunsaturated fatty acids. J Lipid Res 44:479486, 

2003. 

Gao, Z.; Hwang, D.H.; Bataille, F.; Michael, L.; York, D.; Quon, M. J.; Ye, J. 

Serine phosphorylation of insulin receptor substrate 1(IRS-1) by inhibitor Kappa 

B kinase(IKK) complex. J. Biol. Chem. 277:48115-48121, 2002. 

2003 ARS Immunology Research Worskshop 120 
Lee, J.Y.; Hwang, D.H. Docosahexaenoic acid suppresses the activity of 

peroxisome proliferator-activated receptors in a colon tumor cell line. Biochim

Biophys Res Comm. 298:667-674, 2002. 

Han, J.A.; Kim, J.I.; Hwang, D.H.; Ballou, L.R.; Mahale, A.; Aaronson, S.A.; 

Lee, S. W. p53 induction of COX-2 attenuates p53-or genotoxic stress-mediated 

apoptosis EMBO J 21;5635-5644, 2002. 

Paik, J.H.; Lee, J.Y.; Hwang, D. Signaling pathways for TNF-induced COX-2 

expression: mediation through MAP kinases and NFB, and inhibition by certain 

nonsteroidal anti-inflammatory drugs. Advances in Experimental Medicine and 

Biology 57:503-508, 2002. 

Bray, G.A.; Lovejoy, J.C.; Smith, S.R.; Delany, J.P.; Lefevre, M.; Hwang, D.H., 

Ryan, D. H.; York, D.A. The influence of different fats and fatty acids on 

obesity, insulin resistance and inflammation. J. Nutr. 132:2488-2491,2002. 

Hwang, D.H.; Fung, V.; Dannenberg, A.J. National Cancer Institute Workshop 

on Chemopreventive Properties of Non-steroidal Anti-inflammatory Drugs: Role 

of COX-Dependent and –Independent Mechanisms. Neoplasia 4:1-7, 2002. 

Hwang, D.H. Signaling pathways for the expression of cyclooxygenase—

potential targets of modulation by nutrient. FASEB J. 15:2556-2564, 2001. 

Lee, J.Y.; Sohn, K.H.; Rhee, S.H.; Hwang, D.H. Saturated fatty acids, but not 

unsaturated fatty acids, induce the expression of cyclooxygenase-2 mediated 

through Toll-like receptor 4. J Biol Chem 276:16683-16689, 2001. 

Boudreau, M.D.; Sohn, K.H.; Rhee,S.H.; Lee, S.W.; Hunt, J.D.; Hwang, D.H. 

Suppression of tumor cell growth both in nude mice and in culture by n-3 

polyunsaturated fatty acids—mediation through cyclooxygenase-independent 

pathways. Cancer Res 61:1386-1391, 2001. 

Jacob RA

Stephensen, C.B.; Jacob, R.; Aiello, G.; Marquis, G.; Kruzich, L.; Wilson, C.M. 

For the REACH Project. Vitamin C intake and plasma ascorbic acid in HIV+ and 

HIV- U.S. adolescents and young adults in the REACH study. FASEB J 15:A739, 

2001. 

Kelley DS

Benito, P.; Nelson, G. J.; Kelley, D. S.; Schmidt, P. C.; Simon, V. The Effect of 

Conjugated Linoleic Acid on Plasma Lipoproteins and Tissue Fatty Acid 

Composition in Humans. Lipids. 2001. v. 36. pp. 229-236. 

2003 ARS Immunology Research Worskshop 121 
Benito, P.; Nelson, G. J.; Kelley, D. S.; Bartolini, G.; Schmidt, P. C.; Simon, V. 

The Effect of Conjugated Linoleic Acid on platelet Function, Platelet Fatty Acid 

Composition, and Blood Coagulation in Humans. Lipids. 2001. v. 36. pp. 221-

227. 

Hawkes, W.C.; Kelley, D.S.; Taylor, P.C. The effects of dietary selenium on the 

immune system in healthy men. Biol-trace-elem-res. Totowa, N.J. : Humana 

Press. Sept 2001. v. 81 (3) p. 189-213. 

Kelley, D.S.; Simon, V. A.; Taylor, P.C.; Rudolph, I.L.; Benito, P.; Nelson, G.J.; 

Mackey, B.E.; Erickson, K. Dietary Supplementation with Conjugated Linoleic 

Acid Increased Its concentration in Human Peripheral Blood Mononuclear Cells, 

but Did Not Alter Their Function. Lipids. 2001. v. 36. pp. 1-6. 

Kelley, D.S.; Simon, V.A.; Taylor, P.C.; Rudolph, I.L.; Benito, P.; Nelson, G.J.; 

Mackey, B.E.; Erickson, K.L. Dietary supplementation with conjugated linoleic 

acid increased its concentration in human peripheral blood mononuclear cells, but 

did not alter their function. Lipids July 2001. v. 36 (7) p. 669-674. 

Kelley, D.S. Modulation of Human Immune and Inflammatory Responses by 

Dietary Fatty Acids. Nutrition. 2001. v. 17. pp. 669-673. 

Kelley, D.S.; Taylor, P.C.; Rudolph, I.L.; Benito, P.; Nelson, G.J.; Mackey, B.E.; 

Erickson, K. Dietary Conjugated Linoleic Acid Did Not Alter Immune Status in 

Young Healthy women. Lipids. 2000. v. 35. pp. 1065-1071. 

Kelley, D.S.; Warren, J.M.; Simon, V.A.; Bartolini, G.; Mackey, B.E.; Erickson, 

K.L. Similar effects of c9,t11-CLA and t10,c12-CLA on immune cell functions in 

mice. Lipids July 2002. v. 37 (7) p. 725-728. 

Pinna-K; Kelley-D.S.; Taylor, P.C.; King, J.C. Immune functions are maintained 

in healthy men with low zinc intake. Journal-of-Nutrition. 2002, 132: 7, 2033-

2036. 

Rudolph, I.L.; Kelley, D.S.; Klasing, K.C.; Erickson, K.L. Regulation of Cellular 

Differentiation and Apoptosis by Fatty Acids and Their Metabolites. Nutrition 

Research. 2001. v. 21. pp. 381-393. 

Stephensen CB

Drammeh, B.; Marquis, G.S.; Stephensen, C.B.; Eto, I. A randomized fruit-based 

intervention study to improve vitamin A status or rural Gambian children. FASEB 

J 15:A732, 2001. 

2003 ARS Immunology Research Worskshop 122 
Kemper, C.; Leung, M.; Stephensen, C.B.; Pinkert, C.A.; Liszewski, M.K.; 

Cattaneo, R.; Atkinson, J.P. Membrane cofactor protein (MCP; CD46) expression 

in transgenic mice. Clinical and Experimental Immunology 124:180-89, 2001. 

Kruzich, L.A.; Marquis, G.S.; Stephensen, C.B.; Wilson, C.M. for the REACH 

Study. Reported micronutrient intakes among HIV-positive and HIV-negative 

adolescents and young adults in the REACH study. FASEB J 15:A624, 2001. 

Mitra, A.K.; Wahed, M.; Chowdhury, A.; Stephensen, C.B. Factors associated 

with urinary loss of vitamin A in children with watery diarrhea. FASEB J 

15:A256, 2001. 

Stephensen,C.B. Vitamin A, beta-carotene, and mother-to-child transmission of 

HIV. Nutrition-reviews 2003 Aug; 61(8): 280-4. 

Stephensen, C.B. Vitamin A, Infection and Immune Function. Annual Review of 

Nutrition 21:167-92, 2001. 

Stephensen, C.B. Examining the effect of a nutrition intervention on immune 

function in healthy humans: what do we mean by immune function and who is 

really healthy anyway. American Journal of Clinical Nutrition. Nov 2001. v. 74 

(5) p. 565-566. 

Stephensen, C.B.; Jacob, R.; Aiello, G.; Marquis, G.; Kruzich, L.; Wilson, C.M. 

For the REACH Project. Vitamin C intake and plasma ascorbic acid in HIV+ and 

HIV- U.S. adolescents and young adults in the REACH study. FASEB J 15:A739, 

2001. 

Stephensen, C.B.; Rasooly, R.; Jiang, X.; Ceddia, M.A.; Weaver, C.T.; 

Chandraratna, R.A.; Bucy, R.P. Vitamin A enhances in vitro Th2 development via 

retinoid X receptor pathway. Journal-of-immunology 2002 May 1; 168(9): 4495-

503. 

Stephensen, C.B.; Rasooly, R.; Jiang, X.; Ceddia, M.A.; Weaver, C.T.; 

Chandraratna, R.A.S.; Bucy, R.P. Retinoic Acid Modulates in vitro Th1/Th2 

Phenotype Development of Naive T-cells from DO11.10 T-cell Receptor 

Transgenic Mice. FASEB J 15:A1093, 2001. 

Zunino SJ

Kellner, C.; Zunino, S.J. 2003. Nitric oxide is synthesized in response to phenolic 

antioxidants and initially protects against loss of mitochondrial membrane 

potential (DYmt) in leukemia cells. manuscript in preparation. 

Brünke, J., B.; Fischer, K.; Barbin, K.; Schreiter, Y.; Wachter, K.; Mahr, F.; 

Titgemeyer, M.; Peipp; Zunino, S.J.; Repp, R.; Valerius, T.; Fey, G.H.. 2003. A 

2003 ARS Immunology Research Worskshop 123 
recombinant bispecific single-chain Fv antibody against HLA class II and FcgRIII 

(CD16) triggers effective antibody-mediated lysis of lymphoma cells. manuscript 

submitted, Brit. J. Hematol. 

Peipp, M.; Saul, D.; Barbin, K.; Bruenke, J. Zunino, S.J.; van de Winkel, J.G.; 

Niederweis, M.; Fey, G.H.. 2003. Efficient secretion of fluorescent scFv-fusion 

proteins directed against CD antigens from insect and mammalian cells. 

manuscript submitted, J. Immunol. Methods. 

Knott, A.; Mueller, Y.; Zunino, S.J.; Berens, C;. Hillen, W. 2003. Conditional 

cell suicide using dox-dependent caspase-2 expression. J. Gene Med. 5: 343-354. 

Dörrie, J., Sapala, K; . Zunino, S.J. 2002. Interferon-g increases the expression 

of glycosylated CD95 in B-leukemic cells: An inducible model to study the role 

of glycosylation in CD95-signaling and trafficking. Cytokine 18: 98-107. 

Peipp, M.; Küpers, H.; Saul, D.; Schlierf, B.; Greil, J.; Zunino, S.J.; Gramatzki, 

M.; Fey, G.H.. 2002. A recombinant CD7-specific single-chain immunotoxin is a 

potent inducer of apoptosis in acute leukemic T cells. Cancer Res. 62: 2848-

2855. 

Dörrie, J.; Sapala, K.; Zunino, S.J. 2001. Carnosol-induced apoptosis and 

downregulation of Bcl-2 in B-lineage leukemia cells. Cancer Lett. 170: 33-39. 

Dörrie, J.; Gerauer, H.; Wachter, Y.; Zunino, S.J.. 2001. Resveratrol induces 

extensive apoptosis by depolarizing mitochondrial membranes and activating 

caspase-9 in acute lymphoblastic leukemia cells. Cancer Res. 61: 4731-4739. 

Peipp, M.; Simon,, N.; Loichinger, A.; Baum, W.; Mahr, K.; Zunino, S.J., Fey, 

G. H.. 2001. An improved procedure for the generation of recombinant singlechain Fv antibody fragments reacting with human CD13 on intact cells. J. 

Immunol. Methods 251: 161-176. 

Athens, Georgia 

Southeast Poultry Research Laboratory 

http://seprl.ars.usda.gov/

Gast RK

Gast, R.K.; Holt, P.S. The relationship between the magnitude of the specific 

antibody response to experimental Salmonella enteritidis infection in laying hens 

and their production of contaminated eggs. Avian Diseases Apr/June 2001. v. 45 

(2) p. 425-431. 

2003 ARS Immunology Research Worskshop 124 
Gast, R.K.; Nasir, M.S.; Jolley, M.E.; Holt, P.S.; Stone, H.D. Detection of 

experimental Salmonella enteritidis and S. typhimurium infections in laying hens 

by fluorescence polarization assay for egg yolk antibodies. Poultry Science Aug 

2002. v. 81 (8) p. 1128-1131. 

Holt, P.S.; Gast, R.K. Comparison of the effects of infection with Salmonella 

enteritidis, in combination with an induced molt, on serum levels of the acute 

phase protein, alpha1 acid glycoprotein, in hens. Poultry-Science. 2002, 81: 9, 

1295-1300. 

Holt, P.S.; Vaughn, L.E.; Gast, R.K.; Stone, H.D. Development of a lavage 

procedure to collect crop secretions from live chickens for studying crop 

immunity. Avian-Pathology. 2002, 31: 6, 589-592. 

Seo; K.H.; Hol; P.S.; Brackett; R.E.; Gast; R.K.; Stone; H.D. Mucosal humoral 

immunity to experimental Salmonella enteritidis infection in the chicken crop. 

Avian-Diseases. 2002, 46: 4, 1015-1020. 

Seo, K.H.; Holt, P.S.; Vaughn, L.E.; Gast, R.K.; Stone, H.D. Detection of 

Salmonella enteritidis-specific immunoglobulin A antibodies in crop samples 

from chickens infected with Salmonella enteritidis. Poultry-Science. 2003, 82: 1, 

67-70. 

Holt PS

Chaubal, L.H.; Holt,P.S. Characterization of motility and identification of flagella 

proteins in the avian pathogen Salmonella pullorum. American Journal of 

Veterinary Research. 2000. v.60.p.1322-1327. 

Gast, R.K.; Holt, P.S. The relationship between the magnitude of the specific 

antibody response to experimental Salmonella enteritidis infection in laying hens 

and their production of contaminated eggs. Avian Diseases Apr/June 2001. v. 45 

(2) p. 425-431. 

Gast, R.K.; Nasir, M.S.; Jolley, M.E.; Holt, P.S.; Stone, H.D. Detection of 

experimental Salmonella enteritidis and S. typhimurium infections in laying hens 

by fluorescence polarization assay for egg yolk antibodies. Poultry Science Aug 

2002. v. 81 (8) p. 1128-1131. 

Holt, P.S.; Gast, R.K. Comparison of the effects of infection with Salmonella 

enteritidis, in combination with an induced molt, on serum levels of the acute 

phase protein, alpha1 acid glycoprotein, in hens. Poultry-Science. 2002, 81: 9, 

1295-1300. 

2003 ARS Immunology Research Worskshop 125 
Holt, P.S.; Vaughn, L.E.; Gast, R.K.; Stone, H.D. Development of a lavage 

procedure to collect crop secretions from live chickens for studying crop 

immunity. Avian-Pathology. 2002, 31: 6, 589-592. 

Seo, K.H.; Holt, P.S.; Brackett, R.E.; Gast, R.K.; Stone, H.D. Mucosal humoral 

immunity to experimental Salmonella enteritidis infection in the chicken crop. 

Avian-Diseases. 2002, 46: 4, 1015-1020. 

Seo, K.H.; Holt, P.S.; Vaughn, L.E.; Gast, R.K.; Stone, H.D. Detection of 

Salmonella enteritidis-specific immunoglobulin A antibodies in crop samples 

from chickens infected with Salmonella enteritidis. Poultry-Science. 2003, 82: 1, 

67-70. 

Kapczynski DR

Kapczynski, D.R.; Hilt, D.A.; Shapiro, D.; Sellers, H.S.; Jackwood, M.W.

Protection of chickens from infectious bronchitis by in ovo and intramuscular 

vaccination with a DNA vaccine expressing the S1 glycoprotein. Avian Diseases 

2003 Apr-Jun; 47(2): 272-85. 

Kapczynski, D.R.; Sellers, H.S.; Rowland, G.N.; Jackwood, M.W. Detection of 

in ovo-inoculated infectious bronchitis virus by immunohistochemistry and in situ 

hybridization with a riboprobe in epithelial cells of the lung and cloacal bursa.SO: 

Avian Diseases 2002 Jul-Sep; 46(3): 679-85. 

Kapczynski, D.R.; Tumpey, T.M. Development of a virosome vaccine for 

Newcastle disease virus. Avian Diseases 2003 Jul-Sep; 47(3): 578-87. 

Sellers, H.S.; Villegas, P.N.; El-Attrache, J.; Kapczynski, D.R.; Brown, C.C. 

Detection of infectious bursal disease virus in experimentally infected chickens by 

in situ hybridization. Avian Diseases Jan/Mar 2001. v. 45 (1) p. 26-33. 

King DJ

King, D.J. Efficacy of vaccine in protection against velogenic Newcastle Disease 

(ND). Proceedings of the United Animal Health Association.2000. p.591-593. 

King, D.J.; Kommers, G.D.; Brown, C.C.; Seal, B.S. Virulence of pigeon 

Newcastle disease virus isolates for chickens. Proceedings of the Fiftieth Western 

Poultry Disease Conference. 2001.p.15-16. 

King, D.J. Selection of Thermostable Newcastle Disease Virus from Reference 

and Vaccine Strains. Avian Diseases. 2001.v.45.p.512-516. 

2003 ARS Immunology Research Worskshop 126 
King, D.J.; Seal, B.S. Current understanding of molecular correlates of virulence 

in Newcastle Disease Virus. International Hatchery Practice. 2000. 

v.14(7)(Suppl.):Vaccination at work in commercial broilers, p. 10-11. 

King, D.J.; Swayne, D.E. Newcastle disease update: International ND problems. 

Proceedings of the United Animal Health Association. 2000. p. 593-596. 

Kommers, G.D.; King, D.J.; Seal, B.S.; Brown, C.C. Virulence of six 

heterogeneous-origin Newcastle disease virus isolates before and after sequential 

passages in domestic chickens. Avian-Pathology. 2003, 32: 1, 81-93. 

Kommers,G.D.; King,D.J.; Seal, B.S.; Brown, C.C. Pathogenesis of five different 

pigeon-origin isolates of Newcastle disease virus for domestic chickens. 

Veterinary Pathology. 2000.v.37(5).p.546(Abstract No.94). 

Locke, D.P.; Sellers, H.S.; Crawford, J.M.; Schultz-Cherry, S.; King, D.J.; 

Meinersmann, R.J.; Seal, B.S. Newcastle disease virus phosphoprotein gene 

analysis and transcriptional editing in avian cells. Virus Research.2000. v.69.p.55-

68. 

Seal, B.S.; King, D.J. Newcastle Disease Virus. Available from: 

http://www.els.net Encyclopedia of Life Sciences[2000]. 

Sellers, H.S.; Schultz-Cherry, S.; Brown,C.C.; Seal,B.S.; King, D.J. Induction of 

apoptosis by Newcastle disease virus. Program of the Fifth International Congress 

of Veterinary Virology, Brescia, Italy. August,2000.(Abstract). 

Suarez, D.L.;Swayne, D.E.; King, D.J. The ongoing threat of avian influenza and 

Newcastle disease to the U.S. poultry industry. Proceedings of the Fiftieth

Western Poultry Disease Conference.2001.p.1-7. 

Suarez, D.L.; Swayne, D.E.; King, D.J.. The Ongoing Threat of Avian Influenza 

and Newcastle Disease to the US Poultry Industry. Proceedings of the Fiftieth 

Western Poultry Disease Conference. 2001. p.1-7. 

Swayne, D.E.; Suarez, D.L.; King, D.J. Avian influenza (AI) and velogenic 

Newcastle disease (VND) update. Proceedings of 35th National Meeting of 

Poultry Health and Processing.2000.p.37-45. 

Perdue ML

Basler, C.F.; Reid, A.H.; Dybing, J.K.; Janczewski, T.A.; Fanning, T.G.; Zheng, 

H.; Salvatore, M.; Perdue, M.L.; Swayne, D.E.; Garcia-Sastre, A.; Palese, P.; 

Taubenberger, J.K. Sequence of the 1918 Pandemic Influenza Virus Nonstructural Gene (NS) Segment and Characterization of Recombinant Viruses 

Bearing the 1918 NS Genes. Pro. of the Nat'l Acad of Sci 2001. v.98.p.2746-

2751. 

2003 ARS Immunology Research Worskshop 127 
Perdue, M.L.; Suarez, D.L.; Swayne, D.E. Avian influenza in the 90's. Poultry 

and Avian Biology Reviews. 2000. v.11.p.1-20. 

Schultz-Cherry, S.; Dybing, J.K.; Davis, N.L.; Williamson, C.; Suarez, D.L.; 

Johnston, R.; Perdue, M.L. Influenza virus (A/HK/156/97) hemagglutinin 

expressed by an alphavirus replicon system protects chickens against lethal 

infection with Hong Kong-origin H5N1 viruses. 2000. v255.p55-59. 

Swayne, D.E.; Beck,J.R.; Perdue, M.L.; Beard, C.W. Efficacy of Vaccines in 

Chickens Against Highly Pathogenic Hong Kong H5N1 Avian Influenza. Avian 

Diseases. 2001.v.45. p.355-365. 

Seal BS

Cameron, K.; Zhang, X.; Seal, B.; Rodriguez, M.; Njenga, M.K. Antigens to viral 

capsid and non-capsid proteins are present in brain tissues and antibodies in sera 

of Theiler's virus-infected mice. Journal of Virological Methods. 2001.v.91.p.11-

19. 

Gulati, B.R.; Cameron, K.T.; Seal, B.S.; Goyal, S.M.; Halvorson, D.A.; Njenga, 

M.K. Development of highly sensitive and specific enzyme-linked 

immunosorbent assay based on recombinant matrix protein for detection of avian 

pneumovirus antibodies. Journal of Clinical Microbiology. 2000.v.38. p.4010-

4014. 

King, D.J.; Kommers, G.D.; Brown, C.C.; Seal, B.S. Virulence of pigeon 

Newcastle disease virus isolates for chickens. Proceedings of the Fiftieth Western 

Poultry Disease Conference. 2001.p.15-16. 

King, D.J.; Seal, B.S. Current understanding of molecular correlates of virulence 

in Newcastle Disease Virus. International Hatchery Practice. 2000. 

v.14(7)(Suppl.):Vaccination at work in commercial broilers, p. 10-11. 

Kommers, G.D.; King,D.J.; Seal, B.S.; Brown, C.C. Pathogenesis of five different 

pigeon-origin isolates of Newcastle disease virus for domestic chickens. 

Veterinary Pathology. 2000.v.37(5).p.546(Abstract No.94). 

Locke, D.P.; Sellers, H.S.; Crawford, J.M.; Schultz-Cherry, S.; King, D.J.; 

Meinersmann, R.J.; Seal, B.S. Newcastle disease virus phosphoprotein gene 

analysis and transcriptional editing in avian cells. Virus Research.2000. v.69.p.55-

68. 

Seal, B.S.; Sellers, H.S. Avian paramyxoviruses evolve independently of their 

mammalian counterparts and deserve a genus designation among the subfamily 

Paramyxovirinae in the family Paramyxoviridae. Program of the Ninth Annual 

2003 ARS Immunology Research Worskshop 128 
Meeting of the Society for Molecular Biology and Evolution, Athens, GA. July,7-

10, 2001.(Abstract). 

Seal, B.S. Molecular evolution of Newcastle disease virus and the application of 

molecular diagnostics. Program of the Respiratory Diseases of Poultry 

Symposium, American Association of Avian Pathologists at the American 

Veterianry Medical Association Annual Meeting in Boston, MA. July 14-18, 

2001.(Abstract). 

Seal, B.S. Avian Pneumoviruses and Emergence of a New Type in the United 

States. Animal Health Research Reviews.2000.v.1.p.67-72. 

Seal, B.S.; King, D.J. Newcastle Disease Virus. Available from: 

http://www.els.net Encyclopedia of Life Sciences[2000]. 

Sellers, H.S.; Schultz-Cherry,S.; Brown, C.C.; Seal, B.S.; King, D.J. Induction of 

apoptosis by Newcastle disease virus. Program of the Fifth International Congress 

of Veterinary Virology, Brescia, Italy. August,2000.(Abstract). 

Wilson, W.C.; Ma, H.C.; Venter, E.H.; van Djik, A.A., Seal, B.S., Mecham, J.O. 

Erratum to 'Phylogenetic relationships of bluetongue viruses based on gene S7'.

Virus Research 2001. v. 73. p. 201-202. 

Suarez DL

Lee, C.; Song, C.; Lee,Y.; Mo, I.; Garcia, M.; Suarez, D.L.; Kim, S. Molecular 

and pathogenic characterization of Korean isolates of H9N2 avian influenza virus. 

Avian Diseases. 2000. v.44.p.527-535. 

Perdue, M.L.; Suarez, D.L.; Swayne, D.E. Avian influenza in the 90's. Poultry 

and Avian Biology Reviews. 2000. v.11.p.1-20. 

Schultz-Cherry, S.; Dybing, J.K.; Davis, N.L.; Williamson, C.; Suarez, D.L.; 

Johnston, R.; Perdue, M.L. Influenza virus (A/HK/156/97) hemagglutinin 

expressed by an alphavirus replicon system protects chickens against lethal 

infection with Hong Kong-origin H5N1 viruses. 2000. v255.p55-59. 

Suarez, D.L.; Swayne, D.E.; King, D.J. The ongoing threat of avian influenza 

and Newcastle disease to the U.S. poultry industry. Proceedings of the Fiftieth 

Western Poultry Disease Conference.2001.p.1-7. 

Suarez, D.L. Influenza of poultry and swine. Le M,decin V,t,rinaire du Qu,bec. 

2000. v 30.p.187-192. 

2003 ARS Immunology Research Worskshop 129 
Suarez, D.L.; Schultz-Cherry, S. The effect of eukaryotic expression vectors and 

adjuvants on DNA vaccines in chickens using an avian influenza model. Avian 

Diseases. 2000.v 44. p.861-868. 

Suarez, D.L.; Swayne, D.E.; King, D.J.. The Ongoing Threat of Avian Influenza 

and Newcastle Disease to the US Poultry Industry. Proceedings of the Fiftieth 

Western Poultry Disease Conference. 2001. p.1-7. 

Swayne, D.E.; Suarez, D.L.; King, D.J. Avian influenza (AI) and velogenic 

Newcastle disease (VND) update. Proceedings of 35th National Meeting of 

Poultry Health and Processing.2000.p.37-45. 

Swayne, D.E.; Suarez, D.L. Update on International Incidences and Outbreaks of 

Avian Influenza 1999-2000. Proceedings of the US Animal Health Association. 

2000. v.104.p.600-604. 

Swayne DE

King, D.J.; Swayne, D.E. Newcastle disease update: International ND problems. 

Proceedings of the United Animal Health Association. 2000. p. 593-596. 

Perdue, M.L.; Suarez, D.L.; Swayne, D.E. Avian influenza in the 90's. Poultry 

and Avian Biology Reviews. 2000. v.11.p.1-20. 

Perkins, L.E.; Swayne,D.E. Clinicopathologic Aspects of Animal and Zoonotic 

Diseases of Bioterrorism: Avian Influenza. Clinics of North America. 

2001.v.21.p.549-590. 

Perkins, L.E.; Swayne, D.E. Pathobiology of A/Chicken/Hong Kong/220/97 

(H5N1) Avian Influenza Virus in Seven Gallinaceous Species. Veterinary

Pathology. 2001. v.38. p.149-164. 

Suarez, D.L.; Swayne, D.E.; King, D.J. The Ongoing Threat of Avian Influenza 

and Newcastle Disease to the US Poultry Industry. Proceedings of the Fiftieth 

Western Poultry Disease Conference. 2001. p.1-7. 

Swayne, D.E.; Suarez, D.L.; King, D.J. Avian influenza (AI) and velogenic 

Newcastle disease (VND) update. Proceedings of 35th National Meeting of 

Poultry Health and Processing.2000.p.37-45. 

Swayne, D.E. Avian Influenza (AI) and Velogenic Newcastle Disease (VND) 

Update. Proceedings of the National Meeting on Poultry Health and Processing. 

2000. v.35.p.37-45. 

2003 ARS Immunology Research Worskshop 130 
Swayne, D.E. National and International Avian Influenza outbreaks and strategies 

for control. 2001.Proceedings AAAP Symosium on Respiratory Diseases of 

Poultry. p.49-55. 

Swayne, D.E.; Beck, J.R.; Perdue, M.L.; Beard, C.W. Efficacy of Vaccines in 

Chickens Against Highly Pathogenic Hong Kong H5N1 Avian Influenza. Avian 

Diseases. 2001.v.45. p.355-365. 

Swayne, D.E.; Beck, J.R.; Zaki,S. Pathogenicity of West Nile virus for turkeys. 

Avian Diseases. 2000. v.44.p.932-937. 

Swayne, D.E.; Suarez, D.L. Update on International Incidences and Outbreaks of 

Avian Influenza 1999-2000. Proceedings of the US Animal Health Association. 

2000. v.104.p.600-604. 

Tumpey T

Katz, J.M.; Lu, X.; Frace, M.; Morken, T.; Zaki, S.R.; Tumpey, T.M.

Pathogenesis of and Immunity to Avian Influenza A H5 viruses. Biomedicine 

Pharmacotherapy. 2000. v.54.p.178-187. 

Tumpey, T.M.; Lu, X.; Morken, T.; Zaki, S.R.; Katz, J.M. Depletion of

lymphocytes and diminished cytokine production in mice infected with a highly 

virulent influenza A virus isolated from humans. Journal of Virology. 2000. 

v.73.p.6105-6116. 

Tumpey, T.M.; Renshaw, M.; Clements, J.; Katz, J.M. Mucosal delivery of 

inactivated influenza vaccine induces B cell dependent heterosubtypic protection 

against a lethal influenza A H5N1 virus. Journal of Virology. 2001. v.75.p.5141-

5150. 

West Lafayette, Indiana 

Livestock Behavior Research Unit 

http://ag.ansc.purdue.edu/usda/index.html

Eicher SD

Cheng, H.W.; Eicher, S.D.; Chen, Y.; Singleton, P.B.; Muir, W.B. Effect Of 

genetic selection for group productivity and longevity on immunological and 

hematological parameters of chickens. Journal of Poultry Science. 2001. v. 

80:1079-1086. 

Eicher, S.D. Transportation of cattle in the dairy industry: current research and 

future directions. Journal of Dairy Science. 2000. v. 84:19-23. 

2003 ARS Immunology Research Worskshop 131 
Eicher, S.D.; Morrow-Tesch, J.; Albright, J.L.; Williams, R.E. Tail- docking 

alters fly numbers, fly avoidance behaviors and cleanliness, but not physiological 

measures. Journal of Dairy Science. 2001. v. 84:1822- 1828. 

McKee, C.A.; Carroll, J.A.; Eicher, S.D.; Zannelli, M.E.; Beausang, L.A.; 

Matteri, R.L. Supplemental vitamin C and beta-glucan alter growth and the LPSinduced immunological responses in young pigs. Journal of Animal Science. 

2001. v. 79(Suppl.2):Abstract p. 34. 

Cheng HW

Cheng, H.W.; Eicher, S.D.; Chen, Y.; Singleton, P.B.; Muir, W.B. Effect Of 

genetic selection for group productivity and longevity on immunological and 

hematological parameters of chickens. Journal of Poultry Science. 2001. v. 

80:1079-1086. 

West Lafayette, Indiana 

Crop Production and Pest Control Research 

http://www.btny.purdue.edu/USDA-ARS/CPPCRU/CPPCRU.html

Nielsen NC

Helm, R.M.; Cockrell, G.; Connaughton, C.; Sampson, H.A.; Bannon, G.A.; 

Beilinson, V.; Livingstone, D.; Nielsen, N.C.; Burks, A.W. A soybean G2 

glycinin allergen. 1. Identification and characterization. International Archives of 

Allergy and Immunology. 2000, 123: 3, 205-212. 

Helm, R.M.; Cockrell, G.; Connaughton, C.; Sampson, H.A.; Bannon, G.A.; 

Beilinson, V.; Nielsen, N.C.; Burks, A.W. A soybean G2 glycinin allergen. 2. 

Epitope mapping and three-dimensional modeling. International Archives of 

Allergy and Immunology. 2000, 123: 3, 213-219. 

Ames, Iowa 

National Animal Disease Center 

http://www.ars.usda.gov/pandp/locations/locations.htm?modecode=36-25-30-00

Andersen AA 

Everett, K.D.E.; Andersen, A.A. Intergenic Spacer Target Sequence for 

Detection and Distinguishing Chlamydial Strains. 2001. U.S. Patent 6,261,769. 

2003 ARS Immunology Research Worskshop 132 
Sharma, M.; Andersen, A.A. Isolation and growth of Chlamydiae in cell cultures. 

Indian Journal of Animal Sciences. 2000. v. 70(11). p. 1127-1128. 

Bricker BJ

Redkar, R.; Rose, S.; Bricker, B.; DelVecchio, V. Real-time detection of

Brucella abortus, Brucella melitensis and Brucella suis. Molecular Cellular

Probes. 2001. v. 15. P. 43-52. 

Briggs RE

Briggs, R.E. Mucosal Vaccination Against Respiratory Diseases of Ruminants. 

Annual Meeting of the United States Animal Health Association. October 19- 26, 

2000. (Invited Oral Presentation). 

Frank, G.H.; Briggs, R.E.; Duff, G.C.; Loan, R.W.; Purdy, C.W. Effects of 

vaccination prior to transit and administration of florfenicol at time of arrival in a 

feedlot on the health of transported calves and detection of Mannheimia 

haemolytica in nasal secretions. American Journal of Veterinary Research 2002, 

63: 2, 251-256. 

Frank, G.H.; Briggs, R.E.; Loan, R.W.; Purdy, C.W.; Zehr, E.S. Colonization of 

the nasopharynx of cattle by Pasteurella haemolytica: Effect of tilmicosin. 

Haemophilus, Actinobacillus, and Pasteurella International Conference. 1999. 

Abstract p. 10. 

Fulton, R.W.; Purdy, C.W.; Confer, A.W.; Saliki, J.T.; Loan, R.W.; Briggs, R.E.; 

Burge, L.J. Bovine viral diarrhea viral infections in feeder calves with respiratory 

disease: Interactions with Pasteurella spp., parainfluenza-3 virus, and bovine 

respiratory syncytial virus. Canadian Journal of Veterinary Research. 2000. v. 64. 

p. 151-159. 

Jeyaseelan, S.; Kannan, M.S.; Briggs, R.E.; Thumbikat, P.; Maheswaran, S.K. 

Mannheimia haemolytica leukotoxin activates a nonreceptor tyrosine kinase 

signaling cascade in bovine leukocytes, which induces biological effects. 

Infection and Immunity 2001, 69: 10, 6131-6139. 

Jeyaseelan, S.; Kannan, M.S.; Hsuan, S.L.; Walseth, T.F.; Singh, A.K.; Briggs, 

R.E.; Maheswaran, S.K. Role of arachidonic acid metabolites in Pasteurella 

haemolytica leukotoxin-induced cytolysis. Conference for Research Workers in 

Animal Disease. 2000. Abstract #148. 

Leite, F.; Brown, F.J.; Sylte, M.J.; Briggs, R.E.; Czuprynski, C.J. Recombinant 

bovine interleukin-1 beta amplifies the effects of partially purified Pasteurella 

2003 ARS Immunology Research Worskshop 133 
haemolytica leukotoxin on bovine neutrophils in a beta-2-integrin-dependent 

manner. Infection and Immunity. 2000. v. 68. p. 5581-5586. 

Purdy, C.W.; Loan, R.W.; Straus, D.C.; Briggs, R.E.; Frank, G.H. Conglutinin 

and immunoconglutinin titers in stressed calves in a feedlot. American Journal of 

Veterinary Research. 2000. v. 61(11). p. 1403-1409. 

Storz, J.; Lin, M.; Purdy, C.W.; Chouljenko, V.N.; Kousoulas, K.G.; Enright, 

F.M.; Gilmore, W.C.; Briggs, R.E.; Loan, R.W. Coronavirus and Pasteurella 

infections in bovine shipping fever pneumonia and Evans criteria for causation. 

Journal of Clinical Microbiology. 2000. v. 38. p. 3291-3298. 

Thumbikat, P.; Jeyaseelan, S.; Briggs, R.E.; Kannan, M.S.; Maheswaran, S.K. 

Functional dissection of Pasteurella haemolytica leukotoxin mutants. Conference 

for Research Workers in Animal Disease. 2000. Abstract #147. 

Brockmeier SL

Brockmeier, S.L.; Palmer, M.V.; Bolin, S.R. Effects of intranasal inoculation of 

porcine reproductive and respiratory syndrome virus, Bordetella bronchiseptica, 

or a combination of both organisms in pigs. American Journal of Veterinary 

Research. 2000. v. 61(8). p. 892-899. 

Brockmeier, S.L.; Palmer, M.V.; Bolin, S.R.; Rimler, R.B. Effects of intranasal 

inoculation with Bordetella bronchiseptica, porcine reproductive and respiratory 

syndrome virus, or a combination of both organisms on subsequent infection with 

Pasteurella multocida in pigs. American Journal of Veterinary Research. 2001. v. 

62(4). p. 521-525. 

Brockmeier, S.L.; Register, K.B.; Kunkle, R.A. Evaluation of the virulence of a 

dermonecrotic toxin mutant of Bordetella bronchiseptica in swine. Proc. 16th 

International Congress of the Pig Veterinary Society. 2000. p. 480. 

Brockmeier, S.L.; Palmer, M.V.; Bolin, S.R.; Rimler, R.B. Effects of intranasal 

inoculation with Bordetella bronchiseptica, porcine reproductive and respiratory 

syndrome virus, or a combination of both organisms on subsequent infection with 

Pasteurella multocida in pigs. American Journal of Veterinary Research. 2001. v. 

62(4). p. 521-525. 

Register, K.B.; Ducey, T.F.; Brockmeier, S.L.; Dyer, D.W. Reduced virulence of 

a Bordetella bronchiseptica siderophore mutant in neonatal swine. Infection and 

Immunity. 2001. v. 69(4). p.2137-2143. 

2003 ARS Immunology Research Worskshop 134 
Brogden KA

Ackermann, M.R.; Brogden, K.A. Review. Response of the ruminant respiratory 

tract to Mannheimia (Pasteurella) haemolytica. Microbes and Infection. 2000. v. 

2. p. 1079-1088. 

Brogden, K.A.; Kalfa, V.C.; Ackermann, M.R.; Palmquist, D.E.; McCray, P.B. 

Jr.; Tack, B.F. The ovine cathelicidin SMAP29 kills ovine respiratory pathogens 

in vitro and in an ovine model of pulmonary infection. Antimicrobial Agents and 

Chemotherapy. 2001. v. 45. p. 331-334. 

Caverly, J.M.; Radi, Z.A.; Andreasen, C.B.; Doxon, R.A.; Brogden, K.A.;

Ackermann, M.R. Comparison of bronchoalveolar lavage fluid obtained from

Mannheimia haemolytica-inoculated calves with and without prior treatment with 

the selectin inhibitor TBC1269. American Journal of Veterinary Research. 2001. 

v. 62. p. 665-672. 

Lee, H.Y.; Kehrli, M.E. Jr.; Brogden, K.A.; Gallup, J.M.; Ackermann, M.R. 

Influence of beta 2-integrin adhesion molecule expression and pulmonary 

infection with Pasteurella haemolytica on cytokine gene expression in cattle. 

Infection and Immunity. 2000. v. 68(7). p. 4274-4281. 

Radi, Z.A.; Caverly, J.M.; Dixon, R.A.; Brogden, K.A.; Ackermann, M.R. 

Effects of the synthetic selectin inhibitor TBC1269 on tissue damage during acute 

Mannheimia haemolytica-induced pneumonia in neonatal calves. American 

Journal of Veterinary Research. 2001. v. 62. p. 17-22. 

Ramirez-Romero, R.; Brogden, K.A.; Gallup, J.M.; Dixon, R.A.F.; Ackermann, 

M.R. Reduction of pulmonary mast cells in areas of acute inflammation in calves 

with Mannheimia (Pasteurella) haemolytica pneumonia. Journal of Comparative 

Pathology. 2000. v. 123. p. 29-35. 

Zhao, C.; Nguyen, T.; Liu, L.; Sacco, R.E.; Brogden, K.A.; Lehrer, R.I. 

Gallinacin-3, an inducible epithelial beta-defensin of the chicken. 2001. Infection 

and Immunity. v. 69(4). p. 2684-2691. 

Frank GH

Al-Ghamdi, G.M.; Ames, T.R.; Baker, J.C.; Walker, R.; Chase, C.C.L.; Frank 

G.H.; Maheswaran, S.K. Serotyping of Mannheimia (Pasteurella) haemolytica 

isolates from the upper Midwest United States. Journal of Veterinary Diagnostic 

Investigations. 2000. v. 12. p. 576-578. 

Frank, G.H.; Briggs, R.E.; Loan, R.W.; Purdy, C.W.; Zehr, E.S. Colonization of 

the nasopharynx of cattle by Pasteurella haemolytica: Effect of tilmicosin. 

2003 ARS Immunology Research Worskshop 135 
Haemophilus, Actinobacillus, and Pasteurella International Conference. 1999. 

Abstract p. 10. 

Frank, G.H.; Duff, G.C. Effects of tilmicosin phosphate administered prior to 

transport or at time of arrival and feeding of chlortetracycline after arrival in a 

feedlot on Mannheimia haemolytica organisms in nasal secretions of transported 

steers. American Journal of Veterinary Research. 2000. v. 61. p. 1479-1483. 

Purdy, C.W.; Loan, R.W.; Straus, D.C.; Briggs, R.E.; Frank, G.H. Conglutinin 

and immunoconglutinin titers in stressed calves in a feedlot. American Journal of 

Veterinary Research. 2000. v. 61(11). p. 1403-1409. 

Zehr, E.S.; Frank, G.H.;, Tabatabai, L.B. Characterization of Proteins Extracted 

from the Outer Membrane of Mannheimia haemolytica by sepharose- bound 

bovine holotransferrin. 101st General Meeting of the American Society for 

Microbiology. 2001. Abstract #100. 

Harp JA 

Waters, W.R.; Palmer, M.V.; Wannemuehler, M.J.; Sacco, R.E.; Harp, J.A. B 

cells are required for the induction of intestinal inflammatory lesions in TCRαdeficient mice persistently infected with Cryptosporidium parvum. J. Parasitol. 

86:1073-1077. 2000. 

Nonnecke, B.J.; Fowler, M.A.; Pesch, B.A.; Miller, B.L.; Horst, R.L.; Johnson, 

T.E.; Perry, H.B.; Harp, J.A. Function and composition of circulating leukocyte 

populations from milk-replacer-fed, neonatal calves: Effects of dietary vitamins A 

and E. ADSA Annual Mtg., Baltimore, MD, July 24-28, 2000. (Abstract). 

Pesch, B.A; Harp, J.A. Comparative expression of homing receptors on blood 

and milk cells of dairy cattle. Great Lakes International Imaging and Flow 

Cytometry Association Annual Meeting, Detroit MI, October 13-15, 2000. 

(Abstract). 

Smith, J.M.; Nonnecke, B.; Van Amburgh, M.E.; Pesch, B.; Harp, J.A. Effects 

of dietary conjugated linoleic acid (CLA) on the composition and function of 

peripheral blood mononuclear leukocyte populations in heifer calves. 

ADSA/ASAS Annual Mtg., Indianapolis, IN, July 1-5, 2001. (Abstract). 

Harp, J.A.; Stabel, J.R.; Pesch, B.A.; Goff, J.P. Expression of adhesion 

molecules on milk and blood leukocytes from periparturient cows. Conf. Res. 

Workers Anim. Dis. 82:62P. 2001. (Abstract). 

Sonea, I.M.; Schwarz, M.K.; Harp, J.A.;. Regulation of interferon-γ production 

by substance P following infection with Cryptosporidium parvum. Conf. Res. 

Workers Anim. Dis. 82:54P. 2001. (Abstract). 

2003 ARS Immunology Research Worskshop 136 
Heidari, M.; Harp, J.A.; Kehrli, M.E. Expression, purification, and in vitro 

biological activities of recombinant bovine granulocyte colony stimulating factor. 

Vet. Immunol. Immunopathol. 81:45-57. 2001. 

Kehrli, M.E.; Harp, J.A. Immunity in the mammary gland. Vet Clin North Am

Food Anim Pract. 17:495-516. 2001. 

Stabel, J.R.; Goff, J.P.; Kimura, K.; Harp, J.A.; Whitlock, R.H. Effects of

supplemental energy on periparturient immunosuppression in dairy cows with 

Johne’s disease. 7th Intl. Colloquium on Paratuberculosis, Bilboa, Spain, June 

11-14, 2002. (Abstract). 

Harp, J.A.; Marancik, D.; Oesper, J. In vitro comparison of bovine blood and 

milk lymphocyte adherence to high endothelial venules in lymph nodes of murine 

origin. Conf. Res. Workers Anim. Dis. 83:83P. 2002. (Abstract). 

Kimura, K.; Goff, J.P.; Kehrli, M.E.; Harp, J.A.; Nonnecke, B.J. Effects of 

mastectomy on the composition of peripheral blood mononuclear cell populations 

in periparturient dairy cows. J. Dairy Sci. 85:1437-1444. 2002. 

Chen, W.; Harp, J.A.; Harmsen, A.G. Cryptosporidium parvum infection in 

gene targeted B cell deficient mice. J. Parasitol. 89:391-393. 2003. 

Rahner, T.E.; Harp, J.A.; Goff, J.P. Expression of homing receptors on blood and 

milk granulocytes during the parturient period of dairy cattle. Great Lakes 

International Imaging and Flow Cytometry Association Annual Meeting, 

Madison, WI, October 3-5, 2003. (Abstract). 

Harp, J.A.; Oesper J.C.; Casey, T.A. Oral dosing of mice with Escherichia coli 

expressing recombinant interferon gamma reduces Cryptosporidium parvum

infection. Conf. Res. Workers Anim. Dis. 84:93P. 2003. (Abstract). 

Oesper, J.C.; Preszler S.M.; Harp, J.A. Production of recombinant bovine gamma 

interferon by Escherichia coli for use in an oral vaccine to control 

Cryptosporidium parvum infection. Conf. Res. Workers Anim. Dis. 84:91P. 

2003. (Abstract). 

Harp, J.A.; Stabel, J.R.; Rahner, T.E.; Pesch, B.A.; Goff, J.P. Comparison of 

adhesion molecule expression on lymphocytes from blood and milk of normal 

cows and cows with Johne’s disease. Conf. Res. Workers Anim. Dis. 84:71P. 

2003. (Abstract). 

Harp, J.A.; Stabel, J.R.; Pesch, B.A.; Goff, J.P. Expression of adhesion 

molecules on milk and blood leukocytes from periparturient dairy cattle with 

Johne’s disease. Vet. Immunol. Immunopathol. 98:69-76. 2004. 

2003 ARS Immunology Research Worskshop 137 
Harp, J.A.; Waters, T.E.; Goff, J.P. Lymphocyte subsets and adhesion molecule 

expression in milk and blood of periparturient dairy cattle. Vet. Immunol. 

Immunopathol. In press. 2004. 

Horst RL

Nonnecke, B.J.; Waldron, M.R.; Nishida, T.; Overton, T.R.; Horst, R.L.

Circulating leukocyte populations, serum cytokines, and plasma vitamins A and E 

in mid-lactation dairy cows infused with varied doses of lipopolysaccharide. 

International Animal Agriculture and Food Science Conference. 2001. Abstract p. 

385. 

Kehrli ME Jr

Kehrli, M.E. Jr; Harp, J.A. Immunity in the mammary gland. Veterinary Clinics 

of North America, The Food Animal Practice 2001 Nov; 17(3): 495-516. 

Kimura, K.; Goff, J.P.; Kehrli, M.E. Jr.; Harp, J.A.; Nonnecke, B.J. Effects of 

mastectomy on composition of peripheral blood mononuclear cell populations in 

periparturient dairy cows. Journal of Dairy Science June 2002. v. 85 (6) p. 1437-

1444. 

Nonnecke, B.J.; Kimura, K.; Goff, J.P.; Kehrli, M.E. Jr. Effects of the mammary 

gland on functional capacities of blood mononuclear leukocyte populations from

periparturient cows. Journal of Dairy Science 2003 Jul; 86(7): 2359-68. 

Kimura K

Kimura, K.; Goff, J.P.; Kehrli, M.E. Jr.; Harp, J.A.; Nonnecke, B.J. Effects of 

mastectomy on composition of peripheral blood mononuclear cell populations in 

periparturient dairy cows. Journal of Dairy Science June 2002. v. 85 (6) p. 1437-

1444. 

Kimura, K.; Goff, J.P.; Kehrli, M.E. Jr; Reinhardt, T.A. Decreased neutrophil 

function as a cause of retained placenta in dairy cattle. Journal of Dairy Science 

2002 Mar; 85(3): 544-50. 

Nonnecke, B.J.; Kimura, K.; Goff, J.P.; Kehrli, M.E. Jr. Effects of the mammary 

gland on functional capacities of blood mononuclear leukocyte populations from

periparturient cows. Journal of Dairy Science 2003 Jul; 86(7): 2359-68. 

2003 ARS Immunology Research Worskshop 138 
Lager KM

Lager, K.M.; Mengeling, W.L. PRRS: Nature of the RNA virus and how it 

causes disease. Proceedings of International Pig Veterinary Society 16th 

Congress. 2000. p. 538-543. 

Mengeling, W.L.; Bolin, S.R.; Lager, K.M.; Vorwald, A.C.; Wesley, R.D. 

Immunology and epidemiology of porcine reproductive and respiratory syndrome, 

pseudorabies, and postweaning multisystemic wasting syndrome. Proceedings of 

2nd National Meeting of the Italiana Veterinari per Animali da Reddito. 2000. p. 

86-92. 

Mengeling, W.L.; Cutlip, R.C.; Vorwald, A.C.; Lager, K.M. The role of porcine 

reproductive and respiratory syndrome virus and pseudorabies virus in swine 

respiratory disease. Proceedings of 2nd National Meeting of the Italiana 

Veterinari per Animali da Reddito. 2000. p. 93-97. 

Mengeling, W.L.; Lager, K.M.; Vorwald, A.C.; Wesley, R.D.; Prieto, C. Porcine 

reproductive and respiratory syndrome: vaccines and immunity. Magazyn 

Weterynaryjny. 2000. p. 16-18. 

Mengeling, W.L.; Lager, K.M.; Burkhart, K.; Vorwald, A.C.; Roof, M.B.; 

Gorcyca, D.E. Porcine reproductive and respiratory syndrome virus vaccine based 

on isolate JA-142. U.S. Patent Application S/N 09/298,110. 

Mengeling, W.L.; Lager, K.M. Porcine reproductive and respiratory syndrome

(PRRS) in the U.S.A. Proceedings of XVII PanAmerican Congress of Veterinary 

Science. 2000. p. 41-43. 

Mengeling, W.L.; Clouser, D.F.; Vorwald, A.C.; Lager, K.M. Genetic 

recombination among strains of porcine reproductive and respiratory syndrome

virus. Conference of Research Workers in Animal Disease 81st Annual Meeting. 

Abstract No. 134. 

Mengeling, W.L.; Vorwald, A.C.; Cornick, N.A.; Lager, K.M.; Moon, H.W. The 

use of porcine alveolar macrophages for detecting Shiga Toxin. Proceedings of 

American Association of Veterinary Laboratory Diagnosticians 43rd Annual 

Meeting. 2000. p. 82. 

Yaeger, M.; Benson, J.; Christopher-Hennings, J.; Lager, K. Evaluation of the 

sensitivity of fetal serology, IHC, and PCR for the diagnosis of PRRSV in fetuses. 

Proceedings of American Association of Veterinary Laboratory Diagnosticians 

43rd Annual Meeting. 2000. p. 12. 

2003 ARS Immunology Research Worskshop 139 
Lehmkuhl HD

Boyce, W.M.; Woods, L.W.; Keel, K.; MacLachlan, N.J.; Porter, C.O.; 

Lehmkuhl, H.D.; An epizootic of adenovirus induced hemorrhagic disease in 

captive black-tailed deer (Odocoileus hemionus). Journal of Zoo and Wildlife 

Medicine. 2000 31(3). p. 370-373. 

Cutlip, R.C.; Miller, J.M.; Hamir, A.N.; Peters, J.; Robinson, M.M.; Jenny, A.L.; 

Lehmkuhl, H.D.; Taylor, W.D.; Bisplinghoff, F.D. Resistance of cattle to scrapie 

by the oral route. Canadian Journal of Veterinary Research. 2001. v. 65(2). p. 

131-132. 

Debey, B.M.; Lehmkuhl, H.D. Fatal infections in lambs associated with an

adenovirus. 43rd Annual Meeting, American Association of Veterinary 

Laboratory Diagnosticians. 2000. Abstract p. 16. 

Lehmkuhl, H.D.;DeBey, B.M.; Cutlip, R.C. A new serotype adenovirus isolated 

from a goat in the United States. Journal of Veterinary Diagnostic Investigation. 

2001. v. 13(3). p. 195-200. 

Lehmkuhl, H.D.; Woods, L.W. Characterization of a new adenovirus species 

isolated from black-tailed deer in California. Archives of Virology. 2001. v. 

146(6). p. 1187-1196. 

Lehmkuhl, H.D.; DeBey, B.M.; Cutlip, R.C. A new serotype adenovirus isolated 

from a goat in the United States. Journal of Veterinary Diagnostic Investigation. 

2001. v. 13(3). p. 195-200. 

Saliki, J.T; Caseltine, S.L.; Fulton, R.W.; Lehmkuhl H.D. Isolation of bovine 

adenovirus type 7 from calves with respiratory disease. 43rd Annual Meeting, 

American Association of Veterinary Laboratory Diagnosticians. 2000. Abstract p. 

88. 

Woods, L.W.; Swift, P.K.; Lehmkuhl, H.D.; Chiu, P.H.; Hanley, R.S.; 

Nordhausen, R.W.; Stillian, M.H.; Drew, M.L. Experimental adenovirus 

hemorrhagic disease in white-tailed deer fawns. Journal of Wildlife Diseases. 

2001. v. 37(1). p. 153-158. 

Mengeling WL

Lager, K.M.; Mengeling, W.L. PRRS: Nature of the RNA virus and how it 

causes disease. Proceedings of International Pig Veterinary Society 16th 

Congress. 2000. p. 538-543. 

Mengeling, W.L.; Bolin, S.R.; Lager, K.M.; Vorwald, A.C.; Wesley, R.D. 

Immunology and epidemiology of porcine reproductive and respiratory syndrome, 

2003 ARS Immunology Research Worskshop 140 
pseudorabies, and postweaning multisystemic wasting syndrome. Proceedings of 

2nd National Meeting of the Italiana Veterinari per Animali da Reddito. 2000. p. 

86-92. 

Mengeling, W.L.; Cutlip, R.C.; Vorwald, A.C.; Lager, K.M. The role of porcine 

reproductive and respiratory syndrome virus and pseudorabies virus in swine 

respiratory disease. Proceedings of 2nd National Meeting of the Italiana 

Veterinari per Animali da Reddito. 2000. p. 93-97. 

Mengeling, W.L.; Lager, K.M.; Vorwald, A.C.; Wesley, R.D.; Prieto, C. Porcine 

reproductive and respiratory syndrome: vaccines and immunity. Magazyn 

Weterynaryjny. 2000. p. 16-18. 

Mengeling, W.L.; Clouser, D.F.; Vorwald, A.C.; Lager, K.M. Genetic 

recombination among strains of porcine reproductive and respiratory syndrome

virus. Conference of Research Workers in Animal Disease 81st Annual Meeting. 

Abstract No. 134. 

Mengeling, W.L.; Lager, K.M. Porcine reproductive and respiratory syndrome 

(PRRS) in the U.S.A. Proceedings of XVII PanAmerican Congress of Veterinary 

Science. 2000. p. 41-43. 

Mengeling, W.L.; Vorwald, A.C.; Cornick, N.A.; Lager, K.M.; Moon, H.W. The 

use of porcine alveolar macrophages for detecting Shiga Toxin. Proceedings of 

American Association of Veterinary Laboratory Diagnosticians 43rd Annual 

Meeting. 2000. p. 82. 

Mengeling, W.L.; Lager, K.M.; Burkhart, K.; Vorwald, A.C.; Roof, M.B.; 

Gorcyca, D.E. Porcine reproductive and respiratory syndrome virus vaccine based 

on isolate JA-142. U.S. Patent Application S/N 09/298,110. 

Miller JM

Cutlip, R.C.; Miller, J.M.; Hamir, A.N.; Peters, J.; Robinson, M.M.; Jenny, A.L.; 

Lehmkuhl, H.D.; Taylor, W.D.; Bisplinghoff, F.D. Resistance of cattle to scrapie 

by the oral route. Canadian Journal of Veterinary Research. 2001. v. 65(2). p. 

131-132. 

Hamir, A. N.; Cutlip, R.C.; Miller, J.M.; Williams, E.S.; Stack, M.J.; Miller, 

M.W.; O'Rourke, K.I.; Chaplin, M.J. Preliminary findings on the experimental 

transmission of chronic wasting disease agent of mule deer to cattle. Journal of 

Veterinary Diagnostic Investigation. 2001. v13. p.91-96. 

2003 ARS Immunology Research Worskshop 141 
Neill JD

Endsley, J.J.; Ridpath, J.F.; Neill, J.D. Cell mediated immunity to bovine viral 

diarrhea virus in calves protected by maternal antibody. Conference of Research 

Workers in Animal Disease. 2000. Abstract No. 91. 

Leibler-Tenorio, E.M.; Ridpath, J.F.; Greiser-Wilke; Pohlenz, J.F.; Neill, J.D. 

Comparison of lesions and distribution of viral antigen in cattle with acute 

BVDV-2 infection and mucosal disease. European Veterinary Pathology Meeting. 

2000. Abstract No. 31. 

Neill, J.D.; Ridpath, J.F. Recombination with a cellular mRNA encoding a 

DNAJ-like protein results in biotype conversion in genotype 2 bovine viral 

diarrhea viruses. United States-Japan Natural Resources Animal and Avian Health 

Panel. 2000. Abstract No. 11. 

Neill, J.D.; Ridpath, J.F. Further biochemical characterization of three bovine 

capases involved in apoptosis of virus-infected cells. Conference of Research 

Workers in Animal Disease. 2000. Abstract No. 130P. 

Neill, J.D.; Ridpath, J.F. Changes in host cell gene expression following infection 

with bovine viral diarrhea virus genotype 2. American Society for Virology. 2001. 

p. 135. Abstract No. W41-1. 

Ridpath, J.F.; Hietla, S.; Sorden, S.; Neill, J.D. Comparison of diagnostic 

techniques for detection of acute BVDV infections. Conference of Research 

Workers in Animal Disease. 2000. Abstract No. 125P. 

Ridpath, J.F.; Neill, J.D. Characterization of BVDV from North America. United 

States-Japan Natural Resources Animal and Avian Health Panel. 2000. Abstract 

No. 10. 

Ridpath, J.F.; Neill, J.D.; Frey, M.; Landgraf, J.G. Phylogenetic, antigenic and 

clinical characterization of type 2 BVDV from North America. Veterinary 

Microbiology. 2000. v. 77(1-2). p. 145-155. 

Ridpath, J.F.; Neill, J.D.; Endsley, J.; Roth, J.A. Passive immunity and the 

development of a protective response against BVDV. Conference of Research 

Workers in Animal Disease. 2000. Abstract No. 77P. 

Nonnecke BJ

Nonnecke, B.J.; Horst, R.L.; Hammell, D.C.; Franklin, S.T. Effects of

supplemental vitamin A on retinoic acid concentrations in the plasma of 

preruminant calves. International Journal of Vitamin and Nutrition Research. 

2000. v. 70(6). p. 278-286. 

2003 ARS Immunology Research Worskshop 142 
Nonnecke, B.J.; Kimura, K.; Goff, J.P.; Kehrli, M.E. Jr. Effects of the mammary 

gland on functional capacities of blood mononuclear leukocyte populations from

periparturient cows. Journal of Dairy Science 2003 Jul; 86(7): 2359-68. 

Nonnecke, B.J.; Robert, M.P.; Godkin, J.D.; Hammell, D.C.; Franklin, S.T. 

Influence of supplemental, dietary vitamin A on retinol-binding protein 

concentrations in the plasma of preruminant calves. Journal of Dairy Science. 

2001. v. 84. p. 641-648. 

Nonnecke, B.J.; Waldron, M.R.; Nishida, T.; Overton, T.R.; Horst, R.L. 

Circulating leukocyte populations, serum cytokines, and plasma vitamins A and E 

in mid-lactation dairy cows infused with varied doses of lipopolysaccharide. 

International Animal Agriculture and Food Science Conference. 2001. Abstract p. 

385. 

Smith, J.A.; Nonnecke, B.J.; Van Amburgh, M.E.; Pesch, B.A.; Harp, J.A. 

Effects of dietary conjugated linoleic acid (CLA) on the composition and function 

of peripheral blood mononuclear leukocyte populations in heifer calves. 

International Animal Agriculture and Food Science Conference. 2001. Abstract p. 

436. 

Waldron, M.R.; Nishida, T.; Nonnecke, B.J.; Overton, T.R. Metabolic responses 

of lactating dairy cows to intravenous infusion of increasing amounts of 

lipolysaccharide. International Animal Agriculture and Food Science Conference. 

2001. Abstract p. 385. 

Waldron, M.R.; Nishida, T.; Nonnecke, B.J.; Overton, T.R. Hepatic oxidative 

metabolism in lactating cows is modulated by increasing doses of intravenous 

lipopolysaccharide. International Animal Agriculture and Food Science 

Conference. 2001. Abstract p. 385. 

Dean-Nystrom EA

Dean-Nystrom, E.A.; Gansheroff, L.; Twiddy, E.; Moon, H.W.; O'Brien, A.D. 

Passive protection of suckling piglets from Escherichia coli O157:H7 infection by 

vaccination of pregnant sows with intiminO157. International Symposium and 

Workshop on Shiga Toxin (Verotoxin)-Producing Escherichia coli Infections. 

2000. p. 175. 

Dean-Nystrom, E. A.; Melton-Celsa, A. R.; Pohlenz, J. F.; Moon, H. W.; 

O'Brien, A. D. Comparative Pathogenicity of Escherichia coli O157 and IntiminNegative Non-O157 Shiga Toxin-Producing E. coli Strains in Neonatal Pigs. 

Infect Immun. 2003 Nov; 71(11):6526-6533. 

2003 ARS Immunology Research Worskshop 143 
Olsen SC

Adone, R.; Ciuchini, F.; Olsen, S.C. "Antibody responses of heifers vaccinated 

with Brucella abortus RB51 monitored by complement fixation test." 53rd 

Brucellosis Research Conference, Nimess, France. 

Adone, R.; Ciuchini, F.; Olsen S.C. RB51 antigen validation of complement 

fixation test to detect in a blind assay calves of Iowa (USA) vaccinated with 

Brucella abortus RB51. Journal of Clinical Microbiology. 2001. v.8 p. 385-387. 

Capsel, R.L.; Olsen S.C.; Cheville, N.F.; Thoen, C.O. Survival of Brucella 

abortus strain RB51 lyophilized and as a liquid vaccine under different storage 

conditions. Biologicals. 2000. v. 28 p. 209-216. 

Januszewski, M.C.; Olsen, S.C.; McLean, R.G.; Clark, L.; Rhyan, J.C. 

Experimental infection of nontarget species of rodents and birds with Brucella 

abortus strain RB51 vaccine. Journal of Wildlife Diseases. 2001. v. 37. p. 532-

537. 

Olsen, S;C. "Comparison of immune responses to ballistic or hand delivery of 

Brucella abortus strain RB51 vaccine to bison." 2000. Proceedings of the 

Conference on Research Workers in Animal Disease. 

Olsen, S;C. "Update on brucellosis research at NADC" at the Greater 

Yellowstone Interagency Brucellosis Committee Meeting in Jackson, WY on 

November 1, 2000. 

Olsen, S;C.; Lee, I.K. Effect of recombinant interleukin-12 on immune responses 

to brucellosis vaccines. Research Workers in Animal Diseases Conference 

Proceedings. 2000. Abstract No. p. 71. 

Olsen, S.C. Comparison of immune responses to ballistic or hand delivery of 

brucella abortus strain RB51 vaccine to bison. Research Workers in Animal 

Diseases Conference Proceedings. 2000. Abstract No. p. 61. 

Olsen, S;C. "Biosafety Issues in Animal Care and Use" at the American 

Biological Science Safety Association in Washington DC on October 22, 2000. 

Olsen, S;C., Lee, I-K. "Effect of Recombinant Interleukin-12 on Immune 

Responses to Brucellosis Vaccines." Proceedings of the Conference on Research 

Workers in Animal Disease. 

Olsen, S;C. "Immune responses and protection of brucellosis vaccines" at the 

USDA/APHIS/Veterinary Services, Designated Brucellosis Epidemiologist 

Training, Ames, IA, on August 11, 2000. 

2003 ARS Immunology Research Worskshop 144 
Olsen, S;C. "Safety and efficacy of the Brucella abortus strain RB51 vaccine" at 

the 35th Meeting of the United States-Japan Cooperative Program in Natural 

Resources at the National Veterinary Services Laboratory on September 26, 2000. 

Palmer MV

Brockmeier, S.L.; Palmer, M.V.; Bolin, S.R.; Rimler, R.B. Effects of intranasal 

inoculation with Bordetella bronchiseptica, porcine reproductive and respiratory 

syndrome virus, or a combination of both organisms on subsequent infection with 

Pasteurella multocida in pigs. American Journal of Veterinary Research. 2001. v. 

62(4). p. 521-525. 

Brockmeier, S.L.; Palmer, M.V.; Bolin, S.R.; Rimler, R.B. Effects of intranasal 

inoculation with Bordetella bronchiseptica, porcine reproductive and respiratory 

syndrome virus, or a combination of both organisms on subsequent infection with 

Pasteurella multocida in pigs. American Journal of Veterinary Research. 2001. v. 

62(4). p. 521-525. 

Brockmeier, S.L.; Palmer, M.V.; Bolin, S.R. Effects of intranasal inoculation of 

porcine reproductive and respiratory syndrome virus, Bordetella bronchiseptica, 

or a combination of both organisms in pigs. American Journal of Veterinary 

Research. 2000. v. 61(8). p. 892-899. 

Reinhardt TA

Kimura, K.; Goff, J.P.; Reinhardt, T.A.; Sato, S. Association between retained 

placenta and blood interleukin-8 concentration and lack of association of retained 

placenta with energy and calcium metabolic profiles. International Animal 

Agriculture and Food Science Conference. 2001. Abstract p. 151. 

Kimura, K.; Goff, J.P.; Reinhardt. T.A. Parturition blunts calcium signals in 

immune cells. Federation of American Societies for Experimental Biology 

Conference. 2001. Abstract p. 136. 

Register KB

Brockmeier, S.L.; Register, K.B.; Kunkle, R.A. Evaluation of the virulence of a 

dermonecrotic toxin mutant of Bordetella bronchiseptica in swine. Proc. 16th 

International Congress of the Pig Veterinary Society. 2000. p. 480. 

Raffell, T.R.; Register, K.B.; Temple, L.M. Incidence of Bordetella avium in 

wild birds as measured by serology and tracheal cultures. 100th Meeting of the 

American Society for Microbiology. 2000. p. 140. Abstract #C57. 

2003 ARS Immunology Research Worskshop 145 
Register, K.B. RFLP and sequence analysis reveals heterogeneity among the 

pertactin genes of Bordetella bronchiseptica vaccine and field strains. Proceedings 

of the 16th Congress of the International Pig Veterinary Society. 2000. p. 481. 

Register, K.B. Recent advances in understanding the role of Bordetella 

bronchiseptica and its virulence factors in swine respiratory disease. Proc. 16th 

Congress of the International Pig Veterinary Society. 2000. p. 483-488. 

Register, K.B.; Sacco, R.E.; Foster, G. Ribotyping and restriction endonuclease 

analysis reveal a novel clone of Bordetella bronchiseptica in seals. Journal of 

Veterinary Diagnostic Investigation. 2000. v. 12(6). p. 535-540. 

Register, K.B. Novel genetic and phenotypic heterogeneity in Bordetella 

bronchiseptica pertactin. Infection and Immunity. 2001. v. 69(3). p. 1917- 1921. 

Register, K.B.; Ducey, T.F.; Brockmeier, S.L.; Dyer, D.W. Reduced virulence of 

a Bordetella bronchiseptica siderophore mutant in neonatal swine. Infection and 

Immunity. 2001. v. 69(4). p.2137-2143. 

Register, K.B. Genetic polymorphism and its implications in Bordetella 

bronchiseptica. Respiratory Diseases V. Pig Progress. 2001. p. 20-21. 

Sacco, R.E.; Register, K.B.; Nordholm, G.E. Restriction endonuclease analysis 

discriminates Bordetella bronchiseptica isolates. Journal of Clinical 

Microbiology. 2000. v. 38(12). p. 4387-4393. 

Richt JA

Kiermayer, S.; Kraus, I.; Richt, J.A.; Garten, W.; Eickmann, M. Identification of 

the amino terminal subunit of the glycoprotein of Borna disease virus. FEBS 

Letters. 2002 Nov 6; 531(2): 255-8. 

Richt, J.A.; Lager, K.M.; Janke, B.H.; Woods, R.D.; Webster, R.G.; Webby, R.J. 

Pathogenic and antigenic properties of phylogenetically distinct reassortant H3N2 

swine influenza viruses cocirculating in the United States. Journal of Clinical 

Microbiology 2003 Jul; 41(7): 3198-205. 

Mueller, C.A.; Richt, J.A.; Meyermann, R.; Deininger, M.; Schluesener, H. 

Accumulation of the proinflammatory cytokine endothelial-monocyte-activating 

polypeptide II in ramified microglial cells in brains of Borna virus infected Lewis 

rats. Neuroscience Letters 2003 Mar 27; 339(3): 215-8. 

Wolff, T.; Unterstab, G.; Heins, G.; Richt, J.A.; Kann, M. Characterization of an 

unusual importin alpha binding motif in the Borna disease virus p10 protein that 

directs nuclear import. Journal of Biological Chemistry Apr 5, 2002. v. 277 (14) 

p. 12151-12157. 

2003 ARS Immunology Research Worskshop 146 
Ridpath JF

Endsley, J.J.; Ridpath, J.F.; Neill, J.D. Cell mediated immunity to bovine viral 

diarrhea virus in calves protected by maternal antibody. Conference of Research 

Workers in Animal Disease. 2000. Abstract No. 91. 

Flores, E.F.; Gil, L.H.; Botton, S.A.; Weiblen, R.; Ridpath, J.F.; Kreutz, L.C.; 

Pilati, C.; Driemeyer, D.; Moojen, V.; Wendelstein, A.C. Clinical, pathological 

and antigenic aspects of bovine viral diarrhea virus (BVDV) type 2 isolates 

identified in Brazil. Veterinary Microbiology. 2000. v. 77(1-2). p. 175-183. 

Leibler-Tenorio, E.M.; Ridpath, J.F.; Greiser-Wilke; Pohlenz, J.F.; Neill, J.D. 

Comparison of lesions and distribution of viral antigen in cattle with acute 

BVDV-2 infection and mucosal disease. European Veterinary Pathology Meeting. 

2000. Abstract No. 31. 

Neill, J.D.; Ridpath, J.F. Changes in host cell gene expression following 

infection with bovine viral diarrhea virus genotype 2. American Society for 

Virology. 2001. p. 135. Abstract No. W41-1. 

Neill, J.D.; Ridpath, J.F. Further biochemical characterization of three bovine 

capases involved in apoptosis of virus-infected cells. Conference of Research 

Workers in Animal Disease. 2000. Abstract No. 130P. 

Neill, J.D.; Ridpath, J.F. Recombination with a cellular mRNA encoding a 

DNAJ-like protein results in biotype conversion in genotype 2 bovine viral 

diarrhea viruses. United States-Japan Natural Resources Animal and Avian Health 

Panel. 2000. Abstract No. 11. 

Ridpath, J.F.; Neill, J.D.; Endsley, J.; Roth, J.A. Passive immunity and the 

development of a protective response against BVDV. Conference of Research 

Workers in Animal Disease. 2000. Abstract No. 77P. 

Ridpath, J.F.; Neill, J.D.; Frey, M.; Landgraf, J.G. Phylogenetic, antigenic and 

clinical characterization of type 2 BVDV from North America. Veterinary 

Microbiology. 2000. v. 77(1-2). p. 145-155. 

Ridpath, J.F. Detection and molecular typing of ruminant pestiviruses. 

Proceedings of Conference on Pestivirus Contamination of Bovine Sera. 2001. p. 

19-24. 

Ridpath, J.F., Neill, J.D. Characterization of BVDV from North America. United 

States-Japan Natural Resources Animal and Avian Health Panel. 2000. Abstract 

No. 10. 

2003 ARS Immunology Research Worskshop 147 
Ridpath, J.F., Hietla, S., Sorden, S., Neill, J.D. Comparison of diagnostic 

techniques for detection of acute BVDV infections. Conference of Research 

Workers in Animal Disease. 2000. Abstract No. 125P. 

Stoffregen, B.; Bolin, S.R.; Ridpath, J.F.; Pohlenz, J. Morphologic lesions in 

type 2 BVDV infections experimentally induced by strain BVDV2-1373

recovered from a field case. Veterinary Microbiology. 2000. v. 77(1-2). p. 157-

162. 

Van Campen, H.; Ridpath, J.; Williams, E.; Cavender, J.; Edwards, J.; Smith, S.; 

Sawyer, H. Isolation of bovine viral diarrhea virus from a free-ranging mule deer 

in Wyoming. Journal of Wildlife Diseases. 2001. v. 37(2). p. 306-311. 

Vilcek, S.; Ridpath, J.F.; Van Campen, H.; Cavender, J. Identification of a novel 

pestivirus genotype isolated from pronghorn antelope. American Society for 

Virology. 2001. p. 196. Abstract No. P25-9. 

Sacco RE

Waters, W.R.; Palmer ,M.V.; Wannemuehler, M.J.; Sacco, R.E.; Harp, J.A. 2000. B cells 

are required for the induction of intestinal inflammatory lesions of TCR-α-deficient mice 

persistently infected with Cryptosporidium parvum. J. Parasitol. 86:1073-1077. 

Waters, W.R.; Hontecillas, R.; Sacco, R.E.; Zuckermann, F.A.; Harkins, K.R.; 

Bassaganya-Riera, J.; Wannemuehle, M.J. r. 2000. Antigen-specific proliferation of 

porcine CD8αα cells to an extracellular bacterial pathogen. Immunology101:333-341. 

Sacco, R.E., Hutto, D.; Waters, W.R.; Xiasong, L.; Kehrli, M.E. Jr.; Zuckermann,F.; 

Wannemuehler, M.J. 2000. Reduction in inflammation following blockade of CD18 or 

CD29 adhesive pathways during the acute phase of a spirochetal-induced colitis in mice. 

Microb. Pathog. 29:289-299. 

Sonea, I.M.; Jergens, A.E.; Sacco, R.E.; Niyo, Y.; Merten, E.A.; Kaufman, L.A.; Moore, 

P.F. 2000. Flow cytometric analysis of canine colonic and small intestinal lymphocytes 

obtained by endoscopic biopsy in the healthy dog. Vet Immunol. Immunopathol. 77:103-

119. 

Zhao, C.; Nguyen, T.; Liu, L.; Sacco, R.E.; Brogden, K.A.; Lehrer, R.I.. 2001. 

Gallinacin-3, an inducible epithelial β-defensin of the chicken. Infect. Immun. 69:2684-

2691. 

Waters, W.R.; Palmer, M.V.; Sacco, R.E.; Whipple, D.L.. 2002. Nitric oxide production 

as an indication of Mycobacterium bovis infection of white-tailed deer (Odocoileus 

virginianus). J. Wildlife Dis. 38: 338-343. 

2003 ARS Immunology Research Worskshop 148 
Waters, W.R Sacco, R.E.; Fach, S.J.; Palmer, M.V.; Olsen, S.C.; Kreeger, T.J. 2002. 

Analysis of mitogen-stimulated lymphocyte subset proliferation and nitric oxide 

production by peripheral blood mononuclear cells of captive elk (Cervus elaphus). J. 

Wildlife Dis. 38: 344-351. 

Sacco, R.E.; Hagen, M.; Sandor, M.; Weinstock, J.V.; Lynch; R.G. 2002. Established 

Th1 granulomatous responses induced by active Mycobacterium avium infection switch 

to Th2 following challenge with Schistosoma mansoni. Clin. Immunol. 104:274-281. 

.

Brogden, K.A.; Heidari, M.; Sacco, R.E.; Palmquist, D.; Guthmiller, J.M.; Johnson, 

G.K.; Jia, H.P.; McCray, P.B. Jr.; Tack, B.F. 2003. Defensin induced adaptive immunity 

and its potential in preventing periodontal disease. Oral Microbiol. Immunol. 18:95-99. 

Waters, W.R.; Palmer, M.V.; Olsen, S.C.; Sacco, R.E.; Whipple, D.L.. 2003. Immune 

responses of elk to Mycobacterium bovis bacille Calmette Guerin vaccination. Vaccine

21:1518-1526. 

Sacco, R.E., Waters, W.R.; Rudolph, K.M.; Drew, M.L. 2004. Comparative nitric oxide 

production by LPS-stimulated monocyte-derived macrophages from Ovis canadensis and 

Ovis aries. (submitted). 

.

Butler, J.E.; Wertz, N.; Sun, J.; Sacco, R.E.. 2004. Comparison of the expressed porcine 

Vβ and Jβ repertoire of thymocytes and peripheral T cells. (submitted). 

Register, K.B.; Sacco, R.E.; Foster, G. Ribotyping and restriction endonuclease 

analysis reveal a novel clone of Bordetella bronchiseptica in seals. Journal of 

Veterinary Diagnostic Investigation. 2000. v. 12(6). p. 535-540. 

Sacco, R.E.; Register, K.B.; Nordholm, G.E. Restriction endonuclease analysis 

discriminates Bordetella bronchiseptica isolates. Journal of Clinical 

Microbiology. 2000. v. 38(12). p. 4387-4393. 

Waters, W.R.; Hontecillas, R.; Sacco, R.E.; Zuckermann, F.A.; Harkins, K.R.; 

Bassaganya-Riera, J.; Wannemuehler, M.J. Antigen-specific proliferation of 

porcine CD8alpha alpha cells to an extracellular bacterial pathogen. Immunology. 

2000. v. 101(3). p. 333-341. 

Zhao, C.; Nguyen, T.; Liu, L.; Sacco, R.E.; Brogden, K.A.; Lehrer, R.I. 

Gallinacin-3, an inducible epithelial beta-defensin of the chicken. 2001. Infection 

and Immunity. v. 69(4). p. 2684-2691. 

2003 ARS Immunology Research Worskshop 149 
Stabel JR

Dargatz, D.A.; Byrum, B.A.; Barber, L.K.; Sweeney, R.W.; Whitlock, R.H.; 

Shulaw, W.P.; Jacobson, R.H.; Stabel, J.R. Evaluation of a commercial ELISA 

for diagnosis of paratuberculosis n cattle. Journal of American Veterinary 

Medical Association. 2001. v. 218. p. 1163-1166. 

Pillai, S.R.; Jayarao, B.M.; Gummo, J.D.; Gue, E.C.; Tiwari, D.; Stabel, J.R.;

Whitlock, R.H. Identification and sub-typing of Mycobacterium avium subsp. 

paratuberculosis and Mycobacterium avium subsp. avium by randomly amplified 

polymorphic DNA. Veterinary Microbiology. 2001. v. 79. p. 275- 284. 

Stabel, J.R.; Whitlock, R.H. An evaluation of a modified interferon-gamma assay 

for the detection of paratuberculosis in dairy herds. Veterinary Immunology and 

Immunopathology. 2001. v. 79. p. 69-81. 

Stabel, J.R. On-farm batch pasteurization destroys Mycobacterium

paratuberculosis in waste milk. Journal of Dairy Science. 2001. v. 82. p. 524-527. 

Stabel TJ

Fitzsimmons, C.J.; Marklund, L.; Stabel, T.J.; Galina, L.; Tuggle, C.K. 

Evaluation of cross-species hybridization with porcine mRNA to human 

microarrays. International Conference Plant and Genome Research. 2001. 

Abstract p. 212. 

Greiner, L.L.; Stahly, T.S.; Stabel, T.J. The effect of dietary soy genistein on pig 

growth and viral replication during a viral challenge. Journal of Animal Science. 

2001. v. 79. p. 1272-1279. 

Greiner, L.L.; Stahly, T.S.; Stabel, T.J. Quantitative relationship of systemic 

virus concentration on growth and immune response in pigs. Journal of Animal 

Science. 2000. v. 78. p. 2690-2695. 

Meyerholz, D.K.; Stabel, T.J. Preferential cellular adherence of Salmonella 

typhimurium DT104 in a swine gut- loop model. Conference of Research 

Workers in Animal Disease. 2000. Abstract p. 1. 

Stabel, T.J.; Bolin, S.R.; Pesch, B.A. A simple and rapid flow cytometric method 

for detection of porcine cell surface markers. Journal of Immunological Methods. 

2000. v. 245. p. 147-152. 

2003 ARS Immunology Research Worskshop 150 
Stanton TB

Alt, D.P.; Trott, D.J.; Stanton, T.B. Molecular cloning, expression, and DNA 

sequence analysis of a gene encoding an outer membrane protein of Brachyspira 

(Serpulina) pilosicoli. 16th International Pig Veterinary Society. 2000. Abstract p. 

9. 

Li, C.; Corum, L.; Morgan, D.; Rosey, E.L.; Stanton, T.B.; Charon, N.W. The 

spirochete FlaA periplasmic flagellar sheath protein impacts flagellar helicity. 

Journal of Bacteriology. 2000. v. 182. p. 6698-6706. 

Trott, D.J.; Zuerner, R.L.; Wannemuehler, M.; Stanton, T.B. Monoclonal 

antibodies reacting with Brachyspira (Serpulina)pilosicoli membrane proteins. 

16th International Pig Veterinary Society. 2000. Abstract p. 41. 

Tabatabai LB

Bartleson, C.J.; Tabatabai, L.B. Identification of iron-regulated outer membrane 

proteins of Haemophilus parasuis. FASEB Journal. 2001. v. 15(4). p. A533. 

Belzer, C.A.; Tabatabai, L.B.; Frank, G.H. Purification and characterization of 

the Pasteurella haemolytica 35 Kilodalton periplasmic iron-regulated protein. 

Preparative Biochemistry and Biotechnology. 2000. v. 30. p. 343-355. 

Tabatabai, L.B.; Gephart, J.M.; Post, K.W. Two Lys-C endoprotease peptide 

fragments from a major immunoreactive protein from Haemophilus parasuis are 

homologous to H. influenzae elongation Factor Tu (EFTU). FASEB Journal. 

2001. v. 15(4). p. A534. 

Zehr, E.S.; Frank, G.H.; Tabatabai, L.B. Characterization of Proteins Extracted 

from the Outer Membrane of Mannheimia haemolytica by sepharose- bound 

bovine holotransferrin. 101st General Meeting of the American Society for 

Microbiology. 2001. Abstract #100. 

Waters WR

Waters, W.R.; Harp, J.A.; Nonnecke, B.J. 1995. Phenotypic analysis of 

peripheral blood lymphocytes and intestinal intra-epithelial lymphocytes in 

calves. Veterinary Immunology and Immunopathology, 42:249-259. 

Waters, W.R.; Harp, J.A.; Nonnecke, B.J. 1996. Intestinal intraepithelial 

lymphocytes of calves: in vitro blastogenic responses and interferon- 

production. Research in Veterinary Science, 61:45-48. 

Waters, W.R.; Harp, J.A. 1996. Cryptosporidium parvum infection in TCR-- 

and TCR--deficient mice. Infection and Immunity, 64:1854-1857. 

2003 ARS Immunology Research Worskshop 151 
Waters, W.R.; Palmer, M.V.; Ackermann, M.R.; Harp, J.A. 1997. Accelerated 

inflammatory bowel disease of TCR-α-deficient mice persistently infected with 

Cryptosporidium parvum. Journal of Parasitology, 83:460-464. 

Waters, W.R.; Reinhardt, T.A.; Harp, J.A.. 1997. Oral administration of 

putrescine inhibits Cryptosporidium parvum infection of neonatal C57BL-6 mice 

and is independent of nitric oxide synthesis. Journal of Parasitology, 83:746-750. 

Sacco, R.E.; Haynes, J.S.; Harp, J.A.; Waters, W.R.; Wannemuehler, M.J. 1998. 

Cryptosporidium parvum initiates inflammatory bowel disease in germfree T cell 

receptor--deficient mice. American Journal of Pathology, 153:1717-1723. 

Waters, W.R.; Stabel, J.R.; Sacco, R.E.; Harp, J.A.; Pesch, B.A; Wannemuehler, 

M.J. 1999. Antigen-specific B cell unresponsiveness induced by chronic 

Mycobacterium avium subsp. paratuberculosis infection of cattle. Infection and 

Immunity, 67:1593-1598. 

Waters, W.R.; Wannemuehler, M.J.; Sacco, R.E.; Palmer, M.V.; Haynes, J.S.; 

Pesch, B.A.; Harp, J.A.. 1999. Cryptosporidium parvum-induced inflammatory 

bowel disease of TCR-- x TCR--deficient mice. Journal of Parasitology, 

85:1100-1105. 

Waters, W.R.; Sacco, R.E.; Dorn, A.D.; Hontecillas, R.; Zuckermann, F.A.; 

Wannemuehler , M.J. 1999. Systemic and mucosal immune responses of pigs to 

parenteral immunization with a pepsin-digested Serpulina hyodysenteriae 

bacterin. Veterinary Immunology and Immunopathology, 69:75-87. 

Waters, W.R.; Harp, J.A.; Wannemuehler, M.J.; Carbajal, N.Y.; Casas, I.A.. 

1999. Effects of Lactobacillus reuteri on Cryptosporidium parvum infection of 

gnotobiotic TCR--deficient mice. Journal of Eukaryotic Microbiology, 46:60S61S. 

Waters, W.R.; Pesch, B.A.; Hontecillas, R.; Sacco, R.E.; Zuckermann, F.A.; 

Wannemuehler, M.J.. 1999. Cellular immune responses of pigs induced by 

vaccination with either a whole cell sonicate or pepsin-digested Brachyspira 

(Serpulina) hyodysenteriae bacterin. Vaccine, 18:711-719. 

Nonnecke, B.J.; Horst, R.L.; Waters, W.R.; Dubeski, P.; Harp, J.A.. 1999. 

Modulation of fat-soluble vitamin concentrations and blood mononuclear 

leukocyte populations in milk replacer-fed calves by dietary vitamin A and -

carotene. Journal of Dairy Science, 82:2632-2641. 

Thacker, E.L.; Thacker, B.J.; Kuhn, M. P.; Hawkins, A.; Waters, W.R.. 2000. 

Evaluation of local and systemic immune responses induced by intramuscular 

2003 ARS Immunology Research Worskshop 152 
injection of a Mycoplasma hyopneumoniae bacterin to pigs. American Journal of 

Veterinary Research, 61:1384-1389. 

Waters, W.R.; Palmer, M.V.; Wannemuehler, M.J.; Sacco, R.E.; Harp, J.A. 

2000. B cells are required for the induction of intestinal inflammatory lesions of 

TCR-deficient mice persistently infected with Cryptosporidium parvum. Journal 

of Parasitology, 86:1073-1077. 

Sacco, R.E.; Hutto, D.L.; Waters, W.R.; Li, X.; Kehrli, M.E.; Zuckermann, F.A.; 

Wannemuehler, M.J.. 2000. Reduction in inflammation following blockade of 

CD18 or CD29 adhesive pathways during the acute phase of a spirochetalinduced colitis in mice. Microbial Pathogenesis, 29:289-299. 

Waters, W.R.; Hontecillas, R.; Sacco, R.E.; Zuckermann, F.A.; Harkins, K.R.; 

Bassaganya-Riera, J.; Wannemuehler, M.J.. 2000. Antigen-specific proliferation 

of porcine CD8αα cells to an extracellular bacterial pathogen. Immunology, 101: 

333-341. 

Waters, W.R.; Palmer, M.V.; Pesch, B.A.; Olsen, S.C.; Wannemuehler, M.J.; 

Whipple, D.L.. 2000. MHC class II-restricted, CD4+ T cell proliferative 

responses of peripheral blood mononuclear cells from Mycobacterium bovisinfected white-tailed deer. Veterinary Immunology and Immunopathology, 

76:215-229. 

Waters, W.R.; Palmer, M.V.; Pesch, B.A.; Olsen, S.C.; Wannemuehler, M.J.; 

Whipple, D.L.. 2000. Lymphocyte subset proliferative responses of 

Mycobacterium bovis-infected cattle to purified protein derivative. Veterinary 

Immunology and Immunopathology, 77:257-273. 

Waters, W.R.; Nonnecke, B.J.; Rahner, T.E.; Palmer, M.V.; Whipple, D.L.; 

Horst, R.L. 2001. Modulation of Mycobacterium bovis specific responses of 

bovine peripheral blood mononuclear cells by 1,25-dihydroxyvitamin D3. Clinical 

and Diagnostic Laboratory Immunology, 8:1204-1212. 

Palmer, M.V.; Whipple, D.L.; Waters, W.R. 2001. Tuberculin skin testing in 

white-tailed deer (Odocoileus virginianus). Journal of Veterinary Diagnostic 

Investigation, 13:530-533. 

Waters, W.R.; Palmer, M.V.; Sacco, R.E.; Whipple, D.L.. 2002. Nitric oxide 

production as an indication of Mycobacterium bovis infection of white-tailed deer 

(Odocoileus virginianus). Journal of Wildlife Diseases, 38:317-322. 

Waters, W.R.; Sacco, R.E.; Fach, S.J.; Palmer, M.V.; Olsen, S.C.; Kreeger, T.J.. 

2002. Analysis of mitogen-stimulated lymphocyte subset proliferation and nitric 

oxide production by peripheral blood mononuclear cells of captive elk (Cervus 

elaphus). Journal of Wildlife Diseases, 38:323-330. 

2003 ARS Immunology Research Worskshop 153 
Dorn, A.D.; Waters, W.R.; Byers, V.M.; Pesch, B.A.; Wannemuehler, M.J.. 

2002. Characterization of mitogen-stimulated porcine lymphocytes using 

multicolor flow cytometry. Veterinary Immunology and Immunopathology, 87:1-

10. 

Waters, W.R.; Harkins, K.R.; Wannemuehler, M.J.. 2002. Five-color flow 

cytometric analysis of swine lymphocytes for detection of proliferation, apoptosis, 

viability, and phenotype. Cytometry, 48:146-152. 

Waters, W.R.; Palmer, M.V.; Whipple, D.L. 2002. Mycobacterium bovisinfected white-tailed deer (Odocoileus virginianus): detection of immunoglobulin 

specific to crude mycobacterial antigens by ELISA. Journal of Veterinary 

Diagnostic Investigation, 14:470-475. 

Waters W.R.; Rahner, T. E.; Palmer, M. V.; Cheng, D.; Nonnecke, B.J.; 

Whipple, D. L. 2003. Expression of L-selectin (CD62L), CD44, and CD25 on 

activated bovine T cells. Infection and Immunity, 71: 317-326. 

Waters W.R.; Palmer, M.V.; Olsen, S.C.; Sacco, R.E.; Whipple, D.L. 2003. 

Immune responses of elk to Mycobacterium bovis bacille Calmette Guerin 

vaccination. Vaccine, 21: 1518-1526. 

Trott D.J.; Moeller, M.R.; Zuerner, R.L.; Goff , J.; Waters, W.R.; Alt, D.P.; 

Walker, R.L.; Wannemuehler, M.J.. 2003. Isolation, characterization, and immune 

reactivity of Treponema-phagedenis-like spirochetes from papillomatous digital 

dermatitis lesions in dairy cattle. Journal of Clinical Microbiology, 41: 2522-

2529. 

Waters W.R.; Miller, J.M.; Palmer, M.V.; Stabel, J.R.; Jones, D.E.; Koistinen, 

K.A.; Steadham, E.M.; Hamilton, M.J.; Davis, W.C.; Bannantine, J.P. 2003. Early 

induction of a humoral and cellular immune response during experimental 

Mycobacterium avium subsp. paratuberculosis infection of calves. Infection and 

Immunity, 71:5130-5138. 

Nonnecke, B.J.; Waters, W.R.; Foote, M.R.; Fowler, R.M.; Horst, R.L.; Miller, 

B.L. 2003. Interferon- and TNF- secretion by mononuclear leukocytes from

peripheral blood of young and adult cattle vaccinated with Mycobacterium bovis 

BCG: modulation by 1,25-dihydroxyvitamin D3. International Journal of Vitamin 

and Nutrition Research, 73:235-244. 

Waters W.R.; Nonnecke, B.J.; Foote, M.R.; Maue, A.C.; Rahner, T.E.; Palmer, 

M.V.; Whipple, D.L.; Horst, R.L ; Estes, D.M.. 2003. Mycobacterium bovis 

bacille Calmette Guerin vaccination of cattle: activation of bovine CD4+ and  

TCR+ cells and modulation by 1,25-dihydroxyvitamin D3. Tuberculosis, 83:287-

297. 

2003 ARS Immunology Research Worskshop 154 
Waters W.R.; Palmer, M.V.; Whipple, D.L.; Carlson, M.C.; Nonnecke, B.J.. 

2003. Diagnostic implications of antigen-induced IFN-, nitric oxide, and TNF-

 production by blood mononuclear cells from Mycobacterium bovis-infected 

cattle. Clinical and Diagnostic Laboratory Immunology, 10:960-966. 

Waters W.R.; Palmer, M.V.; Nonnecke, B.J ; Whipple, D.L.; Horst, R.L. 2004. 

Mycobacterium bovis infection of vitamin D-deficient NOS2-/- mice. Microbial 

Pathogenesis, 36:11-17. 

Palmer, M.V.; Waters, W.R.; Whipple, D.L.; Slaughter, R.E.; Jones, S.L.. 2004. 

Analysis of interferon- production by Mycobacterium bovis infected whitetailed deer (Odocoileus virginianus) using an in-vitro based assay. Journal of 

Veterinary Diagnostic Investigation, 16:16-21. 

Waters W.R.; Palmer, M.V.; Whipple, D.L.; Slaughter, R.E.; Jones, S.L.. 2004. 

Immune responses of white-tailed deer (Odocoileus virginianus) to 

Mycobacterium bovis BCG vaccination. Journal of Wildlife Diseases, 40:66-78. 

Waters, W.R.; Nonnecke, B.J.; Palmer, M.V.; Robbe-Austermann, S.; 

Bannantine, J.P.; Stabel, J.R.; Whipple, D.L.; Payeur, J.B.; Estes, D.M.; Pitzer, 

J.E.; Minion, F.C.. 2004. Differentiation of Mycobacterium bovis infection of 

cattle from M. avium subsp. avium and M. avium subsp. paratuberculosis 

sensitization using a recombinant ESAT-6:CFP-10 fusion protein. Clinical and 

Diagnostic Laboratory Immunology, in press. 

Palmer, M.V.; Waters W.R.; Pedersen, D.D.; Stoffregen, W.C.. 2004. 

Vaccination of reindeer with a killed West Nile virus vaccine. Journal of Wildlife 

Diseases, in press. 

Endsley, J.J.; Furrer, J.L.; Endsley, M.A.; McIntosh, M.A.; Maue, A.C.; Waters 

,W.R.; Lee, D.R.; Estes, D.M.. 2004. Characterization of a bovine homologue of 

granulysin and NK-lysin with broad spectrum antimicrobial activity. Journal of

Immunology, in press. 

Wesley RD

Mengeling, W.L.; Bolin, S.R.; Lager, K.M.; Vorwald, A.C.; Wesley, R.D.

Immunology and epidemiology of porcine reproductive and respiratory syndrome, 

pseudorabies, and postweaning multisystemic wasting syndrome. Proceedings of 

2nd National Meeting of the Italiana Veterinari per Animali da Reddito. 2000. p. 

86-92. 

Mengeling, W.L.; Lager, K.M.; Vorwald, A.C.; Wesley, R.D.; Prieto, C. Porcine 

reproductive and respiratory syndrome: vaccines and immunity. Magazyn 

Weterynaryjny. 2000. p. 16-18. 

2003 ARS Immunology Research Worskshop 155 
Wesley, R.D. Minimum viral dose for protective lactogenic immunity against 

transmissible gastroenteritis. Proceedings of International Pig Veterinary Society 

16th Congress. 2000. p. 572-573. 

Whipple DL

Palmer, M.V.; Waters, W.R.; Whipple, D.L. Susceptibility of raccoons (Procyon 

lotor) to infection with Mycobacterium bovis. Journal of Wildlife Diseases 2002. 

v. 38 (2) p. 266-274. 

Palmer, M.V.; Whipple, D.L.; Olsen, S.C.; Jacobson, R.H. Cell mediated and 

humoral immune responses of white-tailed deer experimentally infected with 

Mycobacterium bovis. Research in Veterinary Science Feb 2000. v. 68 (1) p. 95-

98. 

Waters, W.R.; Palmer, M.V.; Sacco, R.E.; Whipple, D.L. Nitric oxide production 

as an indication of Mycobacterium bovis infection in white-tailed deer 

(Odocoileus virginianus). Journal of Wildlife Diseases Apr 2002. v. 38 (2) p. 338-

343. 

Waters W.R.; Rahner T.E.; Palmer M.V.; Cheng D.; Nonnecke B.J.; Whipple 

D.L. Expression of L-Selectin (CD62L), CD44, and CD25 on activated bovine T 

cells. Infection and Immunity. 2003, 71: 1, 317-326. 

Whipple, D.L.; Palmer, M.V.; Slaughter, R.E.; Jones, S.L. Comparison of

purified protein derivatives and effect of skin testing on results of a commercial 

gamma interferon assay for diagnosis of tuberculosis in cattle. Journal of 

Veterinary Diagnostic Investigation Mar 2001. v. 13 (2) p. 117-122. 

Yersin AG

Edens, F.W.; Yersin, A.G.; Simmons, D.G. Tryptophan methylester modulation 

of poult responses to Bordetella avium. Poultry Science Mar 1999. v. 78 (3) p. 

327-335. 

Yersin, A.G.; Edens, F.W.; Simmons, D.G. The effects of Bordetella avium

infection on elastin and collagen content of turkey trachea and aorta. Poultry 

Science Nov 1998. v. 77 (11) p. 1654-1660. 

Zuerner RL

Brenot, A.; Trott, D.; Saint Girons, I.; Zuerner, R. Penicillin-binding proteins in 

Leptospira interrogans. Antimicrobial Agents and Chemotherapy. 2001. 45. p. 

870-877. 

2003 ARS Immunology Research Worskshop 156 
Hsu, T.; Hutto, D.L.; Minion, F.C.; Zuerner, R.L.; Wannemuehler, M.J. Cloning 

of a beta-hemolysin gene of Brachyspira (Serpulina) hyodysenteriae and its 

expression in Escherichia coli. Infection and Immunity. 2001. v. 69. p. 706-711. 

Trott, D.J.; Zuerner, R.L.; Wannemuehler, M.; Stanton, T.B. Monoclonal 

antibodies reacting with Brachyspira (Serpulina)pilosicoli membrane proteins. 

16th International Pig Veterinary Society. 2000. Abstract p. 41. 

Zuerner, R.; Haake, D.; Adler, B.; Segers, R. Technological advances in the 

molecular biology of Leptospira. Journal of Molecular Microbiology and 

Biotechnology. 2000. v. 2. p. 455-462. 

New Orleans, Louisiana 

Southern Regional Research Center 

http://www.ars.usda.gov/pandp/locations/locations.htm?modecode=64-35-56-00

Champagne ET

Chung, S.Y.; Maleki, S.J.;Champagne, E.T.; Buhr, K.L.; Gobet, D.W. Peanut 

allergenicity could be enhanced by biochemical reactions occurring during peanut 

roasting. 2000. Journal Allergy Clinical Immunology. v. 105(1), part 2. p. 395. 

Chung, S.Y.; Champagne, E.T. End poducts are ptential cause for the increase in 

IgE-binding of roasted peanuts. American Research Education Society. 2000. 

Abstract p. 24. 

Chung, S.Y.; Butts, C.L.; Champagne, E.T. Effect of curing on peanut 

allergenicity. American Peanut Research and Education Society. 2001. Abstract p. 

21. 

Chung, S.Y.; Champagne, E.T. Association of end-product adducts with 

increased IgE binding of roasted peanuts. Journal Agricultural Food Chemistry. 

2001. v. 49. P. 3592-3597. 

Chung, S.Y.; Champagne, E.T. Effect of peanut maturity on IgE binding. Journal 

Allergy Clinical Immunology. 2001. v. 107. p. 189. 

Maleki, S.J.; Champagne, E.T. Screening the core germplasm collection for 

peanuts with reduced allergenic potential. American Peanut Research Education 

Society. 2001. Abstract p. 47. 

Maleki S.J.; Chung S.Y.; Champagne E.T.; Raufman J.P. The effects of 

processing on the allergenic properties of peanut proteins. Journal Allergy 

Clinical Immunology. 2000. v. 106. p.763-768. 

2003 ARS Immunology Research Worskshop 157 
Maleki, S.J.; Champagne, E.T. Comparing the peanut allergens before and after 

roasting. American Peanut Research Education Society. 2000. Abstract p. 55. 

Maleki, S.J.; Champagne, E.T. Allergenic and biophysical properties of purified 

peanut proteins before and after roasting. Journal Allergy Clinical Immunology. 

2001. v. 103(1), part 2. p. 624. 

Chung S

Chung, S.Y.; Butts, C.L.; Champagne, E.T. Effect of curing on peanut 

allergenicity. American Peanut Research and Education Society. 2001. Abstract p. 

21. 

Chung, S.Y.; Maleki, S.J.; Champagne, E.T.; Buhr, K.L.; Gobet, D.W. Peanut 

allergenicity could be enhanced by biochemical reactions occurring during peanut 

roasting. 2000. Journal Allergy Clinical Immunology. v. 105(1), part 2. p. 395. 

Chung, S.Y.; Champagne, E.T. End poducts are ptential cause for the increase in 

IgE-binding of roasted peanuts. American Research Education Society. 2000. 

Abstract p. 24. 

Chung, S.Y.; Champagne, E.T. Association of end-product adducts with 

increased IgE binding of roasted peanuts. Journal Agricultural Food Chemistry. 

2001. v. 49. P. 3592-3597. 

Chung, S.Y.; Champagne, E.T. Effect of peanut maturity on IgE binding. Journal 

Allergy Clinical Immunology. 2001. v. 107. p. 189. 

Maleki S.J.; Chung S.Y.; Champagne E.T.; Raufman J.P. The effects of 

processing on the allergenic properties of peanut proteins. Journal Allergy 

Clinical Immunology. 2000. v. 106. p.763-768. 

Maleki SJ

Airee, R.; Coke, A.; Airee, S.K.; Maleki, S.J. Correlation between aflatoxins, 

allergens and ingredients of peanuts? American Chemical Society National 

Meeting. 2000. Abstract p.848. 

Bannon, G.A.; Kopper, R.A.; Maleki, S.J.; Shin, D.S.; Sampson, H.A.; Burks, 

A.W. Structure plays a critical role in the allergenicity of food proteins. 2000. 

Journal Allergy Clinical Immunology. V. 105(1), part 2. p. 554. 

Chung, S.Y.; Maleki, S.J.;Champagne, E.T.; Buhr, K.L.; Gobet, D.W. Peanut 

allergenicity could be enhanced by biochemical reactions occurring during peanut 

roasting. 2000. Journal Allergy Clinical Immunology. v. 105(1), part 2. p. 395. 

2003 ARS Immunology Research Worskshop 158 
Maleki S.J.; Chung S.Y.; Champagne E.T.; Raufman J.P. The effects of 

processing on the allergenic properties of peanut proteins. Journal Allergy 

Clinical Immunology. 2000. v. 106. p.763-768. 

Maleki S.J.; Kopper, R. A.; Shin, D.S.; Stanley S.J.; Sampson, H.; Burks A.W.; 

Bannon G.A. Structure of the Major Peanut Allergen Ara h 1 May protect IgEBinding Epitopes form degradation. 2000. Journal Immunology. v. 164. p. 5844-

5849. 

Maleki, S.J.; Champagne, E.T. Comparing the peanut allergens before and after 

roasting. American Peanut Research Education Society. 2000. Abstract p. 55. 

Maleki, S.J.; Champagne, E.T. Allergenic and biophysical properties of purified 

peanut proteins before and after roasting. Journal Allergy Clinical Immunology. 

2001. v. 103(1), part 2. p. 624. 

Maleki, S.J.; Champagne, E.T. Screening the core germplasm collection for 

peanuts with reduced allergenic potential. American Peanut Research Education 

Society. 2001. Abstract p. 47. 

Sen, M.; Kopper, R.A.; Maleki, S.J.; Bannon, G.A.; Sampson, H.A.; Burks, 

A.W.; Bannon, G.A. Structure plays a critical role in the allergenicity of food 

proteins. Journal Allergy Clinical Immunology. 2001. v. 103(1), part 2. p. 612. 

Beltsville, Maryland 

Henry A. Wallace Beltsville Agricultural Research Center 

http://www.ba.ars.usda.gov/

Allen PC

Allen, P.C. Oxidative stress during avian coccidiosis. 8th Coccidiosis 

Conference, 2001. p. 156. 

Allen, P.C.; Fetterer, R.H. Effects of dietary Vitamin E supplementation on 

Eimeria maxima infections in chickens. 8th Coccidiosis Conference.2001. p. 156. 

Zhu, J.; Lillehoj, H.S.; van Tassel, C.; Emara, M.; Allen, P.; Cheng, H.; Pollock, 

D.;, Sadjadi, M.; Sonstegard, T. Mapping QTL loci affecting growth and disease 

resistance to avian coccidiosis. Proceedings of the International Animal 

Agricultural and Food Sciences Conference. 2001. p. 43. 

Zhu, J.; Lillehoj, H. S.; Allen, P.; Cheng, H. H.; Emara, M. G.; Sadjadi, M; 

Pollock, D. 2001. Mapping quantitative trait loci affecting disease resistance to 

2003 ARS Immunology Research Worskshop 159 
avian coccidiosis. Proc. 73rd Northeastern Conference on Avian Diseases, June 

27-29. Virginia- Maryland Regional College of Veterinary Medicine, University 

of Maryland, College Park, MD. p34. 

Zhu, J.; Lillehoj, H. S.; van Tassel, C.; Emara, M.; Allen, P.; Cheng, H.; Pollock, 

D.; Sadjadi, M.; Sonstegard, T. 2001. Mapping QTL loci affecting growth and 

disease resistance to avian coccidiosis. Proc. Int. Animal Agricultural and Food 

Sciences Conference. July 24-28, Indianapolis, IN. p43. 

Zhu, J.; Lillehoj, H.S.; Allen, P.; Cheng, H.H.; Emara, M.G.; Sadjadi, M.; 

Pollock, D. Mapping quantitative trait loci affecting disease resistance to avian 

coccidiosis. Proceedings of the 73rd Northeastern Conference on Avian Diseases. 

2001. p. 34. 

Augustine PC

Augustine, P.C. Eimeria adenoeides sporozoites: enhanced invasion of cultured 

cells previously inoculated with Eimeria acervulina sporozoites. Parasitology 

Research 2001 Jul; 87(7): 548-52. 

Augustine, P.C. Invasion in different cell types by sporozoites of Eimeria species 

and effects of monoclonal antibody 1209-C2 on invasion of cells by sporozoites 

of several apicomplexan parasites. Journal of Eukaryptic Microbiology. 2001. v. 

48. p. 177-181. 

Augustine, P.C. Cell: sporozoite interactions and invasion by apicomplexan 

parasites of the genus Eimeria. International Journal for Parasitology 2001 Jan; 

31(1): 1-8. 

Augustine, P.; Olsen, G.; Danforth, H.; Gee, G.; Novilla, M. Use of monoclonal 

antibodies developed against chicken coccidia (Eimeria) to study invasion and 

development of Eimeria reichenowi in Florida sandhill cranes (Grus canadensis). 

Journal of Zoo and Wildlife Medicine 2001 Mar; 32(1): 65-70. 

Bannerman DD

Bannerman, D.D.; Eiting, K.T., Winn, R.K.; Harlan, J.M. 2004. FLICE-like 

inhibitory protein (FLIP) protects against apoptosis and suppresses NF-κB 

activation induced by bacterial lipopolysaccharide. Am. J. Pathol. In Press.

Bannerman, D.D.; Paape, M.J.; Lee, J.W.; Zhao, X.; Hope, J.C.; Rainard, P. 

2004. Escherichia coli and Staphylococcus aureus elicit differential innate 

immune responses following intramammary infection. Clin. Diagn. Lab. 

Immunol. 11:463-472. 

2003 ARS Immunology Research Worskshop 160 
Bannerman, D.D.; Paape, M.J.; Goff, J.P.; Kimura, K.; Lippolis, J.D.; Hope, 

J.C.. 2004. Innate immune response to intramammary infection with Serratia 

marcescens and Streptococcus uberis. Vet. Res. In press. 

Bannerman, D.D.; Paape, M.J.; Hare, W.R.; Hope, J.C.. 2004. Characterization 

of the bovine innate immune response to intramammary infection with Klebsiella 

pneumoniae. J. Dairy Sci. In press. 

Sakarya, S.; Rifat, S.; Zhou, J.; Bannerman, D.D.; Stamatos, N.M.; Cross, A.S.; 

Goldblum, S.E. 2004. Mobilization of neutrophil sialidase activity desialylates 

the pulmonary vascular endothelial surface and increases resting neutrophil 

adhesion to and migration across the endothelium. Glycobiology. 14:481-494. 

Sohn, E.J.; Paape, M.J.; Peters, R.P.; Fetterer, R.H.; Talbot, N.C.; Bannerman, 

D.D. 2004. The production and characterization of anti-bovine CD14 

monoclonal antibodies. Vet. Res. In press. 

Bannerman, D.D.; Goldblum, S.E.. 2003. Mechanisms of bacterial 

lipopolysaccharide-induced endothelial apoptosis. Am. J. Physiol. Lung. Cell. 

Mol. Physiol. 284:L899-L914. 

Bannerman, D.D.; Paape, M.J.; Hare, W.R.; Sohn, E.J. 2003. Increased levels 

of LPS-binding protein in bovine blood and milk following bacterial 

lipopolysaccharide challenge. J. Dairy Sci. 86:3128-3137. 

Paape, M.J.; Bannerman, D.D.; Zhao, X.; Lee, J.W. 2003. The bovine 

neutrophil: structure and function in blood and milk. Vet. Res. 34:597-627. 

Li, X.; Tupper, J.C.; Bannerman, D.D.; Winn, R.K.; Rhodes, C.J.; Harlan, J.M. 

2003. Phosphoinositide 3 kinase mediates Toll-like receptor 4-induced activation 

of NF-κB in endothelial cells. Infect. Immun. 71:4414-4420. 

Erwert, R.D.; K Eiting,T.; Tupper, J.C.; Winn, R.K.; Harlan, J.M.; Bannerman, 

D.D. 2003. Shiga toxin induces decreased expression of the anti-apoptotic 

protein Mcl-1 concomitant with the onset of endothelial apoptosis. Microb. 

Pathog. 35:87-93. 

Dawson H

Almeria, S.; De-Marez, T.; Dawson, H.; Araujo, R.; Dubey, J.P.; Gasbarre, L.C. 

Cytokine gene expression in dams and foetuses after experimental Neospora 

caninum infection of heifers at 110 days of gestation. Parasite Immunology 2003 

Jul; 25(7): 383-92. 

2003 ARS Immunology Research Worskshop 161 
Dubey JP 

Dubey, J.P.; Garner, M.M.; Stetter, M.D.; Marsh, A.E.; Barr, B.C. Acute 

Sarcocystis falcatula-like infection in a carmine bee-eater (Merops nubicus) and 

immunohistochemical cross reactivity between S. falcatula and Sarcocystis 

neurona. Journal of Parasitology. 2001. v. 87. p. 824-831. 

Kringel, H.; Dubey, J.P.; Beshah, E.; Hecker, R.; Urban, J.F. Effect of CpG 

oligodeoxynucleotide (ODN)-induced immune activation of porcine immunity to 

Toxoplasma gondii. American Association of Veterinary Parasitology. 2001. v. 

46. p. 15. 

Lindsay, D.S.; Dubey, J.P. Direct agglutination test for the detection of 

antibodies to Sarcocystis neurona in experimentally infected animals. Veterinary 

Parasitology. 2001. v. 95. p. 179-186. 

Solano-Aguilar, G.I.; Beshah, E.; Vengroski, K.; Zarlenga, D.; Jauregui, L.; 

Cosio, M.; Douglass, L.W.; Dubey, J.P.; Lunney, J.K. Cytokine and lymphocyte 

profile after oral infection with Toxoplasma gondii oocysts. Scandinavian Journal 

of Immunology. 2001. v. 54. p. 93. 

Solano-Aguilar, G.I.; Beshah, E.; Vengroski, K.; Zarlenga, D.; Jauregui, L.; 

Cosio, M.; Douglass, L.W.; Dubey, J.P.; Lunney, J.K. Cytokine and lymphocyte 

profile in miniature swine after oral infection with Toxoplasma gondii oocysts. 

International Journal of Parasitology. 2001. v. 31. p. 187-195. 

Elsasser TH

Elsasser, T.H.; Klasing, K.C.; Filipov, N.; Thompson, F.; Moberg, G.P. (ed.); 

Mench, J.A. The metabolic consequences of stress: targets for stress and priorities 

of nutrient use. The biology of animal stress: basic principles and implications for 

animal welfare. 2000, 77-110. CABI Publishing; Wallingford; UK. 

Kahl, S.; Elsasser, T.H.; Sartin, J.L.; Fayer, R. Effect of progressive cachectic 

parasitism and growth hormone treatment on hepatic 5'-deiodinase activity in 

calves. Domestic Animal Endocrinology June 2002. v. 22 (4) p. 211-221. 

Perkins, K.H.; VandeHaar, M.J.; Burton, J.L.; Liesman, J.S.; Erskine, R.J.; 

Elsasser, T.H. Clinical responses to intramammary endotoxin infusion in dairy 

cows subjected to feed restriction. Journal of Dairy Science July 2002. v. 85 (7) p. 

1724-1731. 

Pio, R.; Martinez, A.; Elsasser, T.H.; Cuttitta, F. Presence of immunoreactive 

adrenomedullin in human and bovine milk. Peptides. 2000. v. 21(12). p. 1859-63. 

2003 ARS Immunology Research Worskshop 162 
Richards, M.P.; Caperna, T.J.; Elsasser, T.H.; Ashwell, C.M.; McMurtry, J.P. 

Design and application of a polyclonal peptide antiserum for the universal 

detection of leptin protein. Journal of Biochemical and Biophysical Methods. 

2000. v. 45. p. 147-156. 

Evans JD

Evans, J.D. Microsatellite loci in the honey bee parasitic mite Varroa jocabsoni. 

Molecular Ecology. 2000. v. 9. p. 1436-1438. 

Fayer R

Higgins, J.; Jenkins, M.C.; Fayer, R.; Xiao, L.; Lal, A.A.; Bennet, B.; Richards, 

J. New methods for molecular diagnostics for emerging waterborne pathogens. 

Proceedings of the International Conference on Emerging and Infectious

Diseases. p. 38. 

Jenkins, M.C.; Trout, J.; Higgins, J.; Fayer, R. The mRNA levels for 

amyloglucosidase as measured by RT-PCR correlate with infectivity and 

fecundity of Cryptosporidium parvum oocysts for neonatal BALB/c mice. 

Proceedings of the International Conference on Emerging and Infectious

Diseases. Late Breakers. 2000. p. 9. 

Jenkins, M.C.; Trout, J.; Murphy, C.; Harp, J.; Higgins, J.; Wergin, W.; Fayer, R. 

Cloning and expression of a DNA sequence encoding a 41-kilodalton 

Cryptosporidium parvum oocyst wall protein. Clinical and Diagnostic Laboratory 

Immunology. 1999. v. 6(6). p. 912-920. 

Fetterer RH

Allen, P.C.; Fetterer, R.H. Effects of dietary Vitamin E supplementation on 

Eimeria maxima infections in chickens. 8th Coccidiosis Conference.2001. p. 156. 

Allen, P.C.; Fetterer, R.H. Effect of Eimeria acervulina infections on plasma larginine. Poultry Science. 2000. v. 79. p. 1414-1417. 

Fetterer, R.H.; Allen, P.C. Eimeria acervulina elevates plasma and muscle 3-

methylhistindine levels in chickens. Journal of Parasitology. 2000. v 86. p. 783-

781. 

Gasbarre LC

Gasbarre, L.C.; Stout, W.L.; Lohr, L.; Guerrero, J. Gastrointestinal nematodes 

on a grazing dairy operation: Effect of parasite control on production and parasite 

transmission. Proceedings of the American Association of Veterinary 

Parasitology. 2001. p. 51. 

2003 ARS Immunology Research Worskshop 163 
Gasbarre, L.C.; Leighton, E.A.; Sonstegard, T. Role of the bovine immune system

and genome in resistance to gastrointestinal nematodes. Veterinary Parasitology. 

2001. v. 98. p. 51-64. 

Hediger-Weithaler, B.M.; Heussler, V.T.; Eicher, R.; Zakher, A.; Zarlenga, D.S.; 

Gasbarre, L.C.; Waldvogel, A.S. Interferon- and interleukin 4 mRNA expression 

by PBMCs from BVDV seropositive pregnant and non-pregnant cattle. Veterinary 

Immunology and Immunopathology. 2000. v. 77. p. 201-212. 

Sonstegard, T.S.; Gasbarre, L.C.; Van Tassell, C.P.; Padilha, T. Genomic tools 

to improve parasite resistance. Proceedings of the American Association of

Veterinary Parasitologists. 2001. v. 46. p. 46. 

Zarlenga, D.S.; Chute, M.B.; Gasbarre, L.C.; Boyd, P. A multiplex PCR assay 

for differentiating economically important gastrointestinal nematodes of cattle. 

Veterinary Parasitology. 2001. v. 97. p. 199-209. 

Jenkins MC

Baker, S.A.; Jenkins, M.; Chappell, C.L. Validating a recombinant protein 

antigen in a serodiagnostic test for antibodies to Cryptosporidium parvum. 

Proceedings of the 51st Annual Meeting of the American Society of 

Parasitologists, Albuquerque, New Mexico. 2001. p. 50. 

Dyer, R.M.; Jenkins, M.C.; Kwok, O.C.H.; Douglas, L.W.; Dubey, J.P. 

Serologic survey of Neospora caninum infection in a closed dairy cattle herd in 

Maryland: risk of serologic reactivity by production groups. Veterinary 

Parasitology. 2000. v. 90. p. 171-181. 

Fayer, R.; Lewis, E.J.; Trout, J.M.; Graczyk, T.K.; Jenkins, M.C.; Higgins, J.; 

Xiao, L.; Lal, A.A. Cryptosporidium parvum in oysters from commercial 

harvesting sites in the Chesapeake Bay. Emerging Infectious Diseases. 1999. v. 5. 

p. 706-710. 

Higgins, J.; Jenkins, M.C.; Fayer, R.; Xiao, L.; Lal, A.A.; Bennet, B.; Richards, 

J. New methods for molecular diagnostics for emerging waterborne pathogens. 

Proceedings of the International Conference on Emerging and Infectious

Diseases. p. 38. 

Jenkins, M.C.; Trout, J.; Murphy, C.; Harp, J.; Higgins, J.; Wergin, W.; Fayer, R. 

Cloning and expression of a DNA sequence encoding a 41-kilodalton 

Cryptosporidium parvum oocyst wall protein. Clinical and Diagnostic Laboratory 

Immunology. 1999. v. 6(6). p. 912-920. 

2003 ARS Immunology Research Worskshop 164 
Jenkins, M.C.; Botero, S.; Allen, P.; Danforth, H. Application of non- attenuated 

and gamma-irradiated eimerian oocysts to control avian coccidiosis. Proceedings 

of the Annual Meeting of the American Association of Veterinary Parasitologists. 

p. 85. 

Jenkins, M.C.; Trout, J.; Higgins, J.; Fayer, R. The mRNA levels for 

amyloglucosidase as measured by RT-PCR correlate with infectivity and 

fecundity of Cryptosporidium parvum oocysts for neonatal BALB/c mice. 

Proceedings of the International Conference on Emerging and Infectious

Diseases. Late Breakers. 2000. p. 9. 

Jenkins, M.C.; Caver, J.A.; Bjorkman, C.; Anderson, T.C.; Romand, S.; Vinyard; 

Uggla, A.; Thulliez, P.; Dubey, J.P. Serological investigation of an outbreak of 

Neospora caninum-associated abortion in a dairy herd in southeastern United 

States. Veterinary Parasitology. 2000. v. 94. p. 17- 26. 

Jenkins, M.C.; Trout, J.; Abrahamsen, M.S.; Lancto, C.A.; Higgins, J.; Fayer, R. 

Estimating viability of Cryptosporidium parvum oocysts using reverse 

transcriptase-polymerase chain reaction (RT-PCR) directed at mRNA encoding 

amyloglucosidase. Journal of Microbial Methods. 2000. v. 43. p. 97-106. 

Jenkins, M.C.; Trout, J.; Fayer, R. Treatment of Cryptosporidium parvum 

sporozoites with antibodies to a recombinant CP47 surface antigen inhibits in 

vitro parasite development. Proceedings of the Annual Meeting of the American 

Association of Veterinary Parasitologists. Boston, Massachusetts. 2001. p. 49. 

Lillehoj, H.S.; Jenkins, M.C.; Vakharia, V. Eimeria protein inducing interferonproduction. Comparison of different gene expression systems and immunization 

strategies for vaccination against coccidiosis. World Poultry Congress. 2000. p. 

17-13. 

Lillehoj, H.S.; Choi, K.D.; Jenkins, M.C.; Vakharia, V.N.; Song, K.D.; Han, 

J.Y.; Lillehoj, E.P. A recombinant Eimeria protein inducing interferonproduction. Comparison of different gene expression systems and immunization 

strategies for vaccination against coccidiosis. Avian Diseases. 2000. v. 44. p. 379-

389. 

Lillehoj HS

Lillehoj, H. S.; Lillehoj, E. P. 2000. Avian coccidiosis. A review of acquired 

intestinal immunity and vaccination strategies. Avian Dis. 44:408-425. 

Lillehoj, E. P.; Yun, C. H.; Lillehoj, H. S. 2000. Vaccine for Avian 

enteropathogens Eimeria, Cryptosporidium and Salmonella. Animal Health 

Research Reviews. 1:47-65. 

2003 ARS Immunology Research Worskshop 165 
Lillehoj, H. S.; Jenkins, M.C.; Vakharia, V. 2000. Eimeria protein inducing 

interferon-γ production. Comparison of different gene expression systems and 

immunization strategies for vaccination against coccidiosis. In “Proc. World 

Poultry Congress,” P17.13. (CD). 

Lillehoj, H. S.; Choi, K. D.; Jenkins, M.C.; Vakharia, V.N.; Song, K. D.; Han, J. 

Y.; Lillehoj, E. P. 2000. A recombinant Eimeria protein inducing interferon-γ

production. Comparison of different gene expression systems and immunization 

strategies for vaccination against coccidiosis. Avian Dis. 44:379-389. 

Lillehoj, H. S. 2000. Mucosal Immune response to coccidiosis. In “Proc. XXI 

World Poultry Congress”. S1.10. (CD). 

Lillehoj, H. S. 2000. Review on vaccine development against enteric parasites 

Eimeria and Cryptosporidium. Japanese. Poul. Sci. 37: 117-141. 

Lillehoj, H. S. 2000. Subcutaneous injection with IL-2 gene enhances cellmediated immunity. In "Proc World Poultry Congress," P17.14. (CD). 

Yun, C. H.; Lillehoj, H. S.. 2000. Intestinal immune responses to coccidiosis. 

Dev. Comp. Immunol. 24:303-324. 

Choi, K. D.; Lillehoj, H. S. 2000. Effect of chicken IL-2 on T-cell growth and 

function. Increased T cell response following DNA immunization. Vet Immunol 

Immunopathol. 73:309-312. 

Yun, C. H.; Lillehoj, H. S.; Choi, K. D. 2000. Chicken IFN-γ monoclonal 

antibodies and their application in enzyme-linked immunosorbent assay. Vet. 

Immunol. Immunopathol. 73:297-308. 

Yun, C. H.; Lillehoj, H. S.; Choi, K. D. 2000. Eimeria tenella infection induces 

local IFN-γ production and intestinal lymphocyte subpopulation changes. Infect. 

Immunity. 68: 1282-1288. 

Yun, C. H.; Lillehoj, H. S.; Zhu, J.; Min, W.G. 2000. Kinetic differences in 

intestinal and systemic interferon-γ and antigen-specific antibodies in chickens 

infected with Eimeria maxima. Avian Dis. 44:305-312. 

Zhu, J.; Lillehoj, H. S.; Allen, P.; Yun, C. H.; Pollock, D.; Emara, M. 2000. 

Analysis of disease resistance associated parameters in broiler chickens 

challenged with Eimeria maxima. Poul. Sci. 79:619-625. 

Song, K. D.; Lillehoj, H. S.; Choi, K. D.; Parcells, M. S.; Huynh, J. T.; Yun, C. 

H.; Han, J.Y. 2000. A DNA vaccine encoding a conserved Eimeria protein 

induces protective immunity against live Eimeria acervulina challenge. Vaccine. 

19:243-252. 

2003 ARS Immunology Research Worskshop 166 
Sasai, K.; Aita. M.; Lillehoj, H. S.; Miyamoto, T.; Fukata, T.; Baba, E. 2000. 

Dynamics of lymphocyte subpopulation changes in the cecal tonsils of chickens 

infected with Salmonella enteritidis. Vet. Microbiol. 74:345-351. 

Kliger, C. A.; Gehad, A. E.; Hulet, R. M.; Roush, W. B.; Lillehoj, H. S.; 

Mashaly, M. M. 2000. Effect of melatonin and photoperiod on broiler immune 

system. Poultry Sci. 79:18-25. 

 Min, W.; Lillehoj, H. S. 2000. Synthetic construct for anti-Eimeria acervulina 

surface ScFv antibody, clone 5D11. Genbank, Accession Number. AJ298108. 

Min, W.; Lillehoj, H. S. 2000. Synthetic construct for anti-Eimeria acervulina 

surface ScFv antibody, clone 2-1. Genbank, Accession Number. AJ298107. 

Lillehoj, H. S.; Yun, C. H.; Nichols, M. 2000. Monoclonal antibodies detecting 

chicken IFN-gamma and development of in vitro assay. Patent Serial Number 

09/534,002, Document Number 0165.99. 

Lillehoj, H. S. 2001. Application of DNA marker Technology to the development 

of coccidiosis control strategy. In “Poultry Biotechnology and its Applications”, 

Proceeding of Korean Society of Poultry Science Symposium. pp1-17, Suwon, 

Korea. 

Dalloul, R.A.; Lillehoj, H. S.; Doerr, J.A. 2001. Probiotics in Broilers: 

Modulation of Intestinal Immune Function. In: Proc. 48th Md. Nutr. Conf. Feed 

Manuf. Pages 30-41. 

Lillehoj, H. S.; Min, W.; Choi, K. D.; Babu, U. S.; Burnside, J.; Miyamoto, T.; 

Rosenthal, B. M.; Lillehoj, E. P. 2001. Molecular, cellular, and functional 

characterization of chicken cytokines homologous to mammalian IL-15 and IL-2. 

Vet. Immunol. Immunopath. 82: 229-244. 

Lillehoj, H. S.; Min, W. G.; Choi, K. D.; Babu, U.; Burnside, J.; Miyamoto, T.; 

Lillehoj, E. P. 2001. Functional Characterization of Chicken Cytokines 

Homologous to Mammalian IL-15 and IL-2. In "Current Progress on Avian 

Immunology Research," K. A. Schat ed, American Association of Avian 

Pathologists, pp8-13, Kennett Square, PA. 

Lillehoj, H. S.; Zhu, J.; Min, W.G. 2001. Application of DNA marker 

Technology and functional genomics to the development of coccidiosis control 

strategy. Korean J. Poultry Sci 28:107-123. 

Guangxing, L.; Lillehoj, E.; Lillehoj, H. S. 2001. Interleukin-2 production in SC 

and TK chickens infected with Eimeria tenella. Avian Diseases. 46:2-9. 

2003 ARS Immunology Research Worskshop 167 
Guangxing L.; Lillehoj, H. S.; Min, W. 2001. Production and Characterization of 

Monoclonal Antibodies Detecting the Chicken Interleukin-15 Receptor Alpha 

Chain. Vet. Immunol. Immunopathol. 82: 215-227. 

Gehad A. E.; Lillehoj, H. S.; Mashaly, M. 2001. The role of lymphocytes and 

hormones during the development of the delayed hypersensitivity response in 

immature male chickens. In "Current Progress on Avian Immunology Research," 

K. A. Schat, ed., American association of Avian Pathologists, pp308-314, Kennett 

Square, PA. 

Kim, J. K.; Lillehoj, H. S.; Min, W. G.; Han, J. Y.; Sasai, K.; Sohn, E. J.; 

Lillehoj, E. P. 2001. Recombinant Single Chain Fragment Variable Region 

Antibody Constructed from Chicken Monoclonal Antibody Detects Apical 

Complex Protein of Eimeria acervulina. In "Current Progress on Avian 

Immunology Research," K. A. Schat, ed., American Association of Avian 

Pathologists, pp 64-70, Kennett Square, PA. 

Min, W. G.; Lillehoj, H. S.; Burnside, J.; Weining, K. C.; Staeheli, P.; Nichols, M 

B. 2001. Effect of IL-1, IL-8, IL-15, IL-2, IFN-α, IFN-γ, Lymphotactin and TGFβ4 on DNA vaccination against Eimeria acervulina. In "Current Progress on 

Avian Immunology Research," K. A. Schat, ed, American Association of Avian 

Pathologists, pp352-356, Kennett Square, PA. 

Zhu, J.; Lillehoj, H. S.; Cheng, H. H.; Pollock, D.; Sadjadi M.; Emara, M. 2001. 

Screening for highly heterozygous chickens in outbred commercial broiler lines to 

increase detection power for mapping quantitative trait loci. Poul Sci. 80:6-21. 

Miyamoto, T.; Lillehoj, H. S.; Sohn, E. J.; Min, W. 2001. Production and 

characterization of monoclonal antibodies detecting chicken interleukin-2 and the 

development of an antigen capture enzyme-linked immunosorbent assay. Vet. 

Immunol. Immunopathol. 80:245-257. 

Min, W. G.; Lillehoj, H. S.; Burnside, J.; Kim, H. B.; Lillehoj, E.. 2001. Northern 

blot analysis of concanavalin A-induced expression of chicken gene homologous 

to the mammalian IL-2 receptor chain. In "Current Progress on Avian 

Immunology Research," Ed. K. A. Schat, American Association of Avian 

Pathologists, pp346-351, Kennett Square, PA. 

Min, W.; Kim, J. K.; Lillehoj, H. S.; Sohn, E. J.; Han, J. Y.; Song, K. D.; Lillehoj 

E. 2001. Characterization of recombinant scFv antibody reactive with an apical 

antigen of Eimeria acervulina. Biotechnology Letters 23:949-955. 

Kim, J. K.; Min, W.; Lillehoj, H. S.; Kim, S. W.; Sohn, E. J.; Song, K. D.; Han, J. 

Y. 2001. Generation and characterization of recombinant scFv antibodies 

detecting Eimeria acervulina surface antigens. Hybridoma 20:175-181.

2003 ARS Immunology Research Worskshop 168 
Miyamoto, T.; Min, W.; Lillehoj, H. S. 2001. Lymphocyte proliferation response 

during Eimeria tenella infection assessed by a new, reliable, non-radioactive 

colorimetric assay. Avian Diseases 46:10-16. 

Miyamoto, T.; Min, W.; Lillehoj, H. S. 2001. Kinetics of interleukin-2 

production in chickens infected with Eimeria tenella. Comparative Immunology, 

Microbiology and Infectious Diseases, 25:149-158. 

Song, K. D.; Han, J. Y.; Min, W.; Lillehoj, H. S.; Kim, S.; Kim, J.2001. 

Molecular cloning and characterization of cDNA encoding immunoglobulin 

heavy and light chain variable regions from four chicken monoclonal antibodies 

specific to surface antigens of intestinal parasites, Eimeria acervulina. The Korean 

J. Microbiology. 39(1): 49-55. 

Hwang, K. C.; Song, K. D.; Kim, T. H.; Lee, H. K.; Sohn, S. H.; Lillehoj, H. S.; 

Han, J. Y. 2001. Genetic linkage mapping of RAPD markers segregating in 

Korean Ogol chickens-White Leghorn backcross population. Asian-Aust. J. 

Anim. Sci. 14:302-306. 

Min, W.; Kim, J. K.; Lillehoj, H. S.; Sohn, E. J.; Han, J. Y.; Song, K. D.; 

Lillehoj, E. P. 2001. Synthetic construct for anti-Eimeria acervulina apical protein 

antibody, clone 6D12. Genbank. Accession Number. AJ3000836. 

Zhang, S.; Lillehoj, H. S.; Jenkins, M. C.; Lally, N.; Min, W. 2001. Gallus gallus 

mRNA for putative FK506-binding protein (FKBP25 gene). Genbank, Accession 

Numbr. AJ312905. 

Lillehoj, H. S. 2002. Beyond genomics revolution: Emergence of agricultural 

biotechnology. In “Proceeding of Future Perspectives in Animal Biotechnology,” 

pp 5-21, Seoul National University, College of Animal Science and Technology, 

Suwon, Korea. 

Lillehoj, H. S. 2002. Gut immune responses and intervention strategies against 

Eimeria and Salmonella. In “Proceeding of International Seminars on Avian 

Pathology and Production,” Pp 91-131, Chile, Santiago. 

Lillehoj, H. S. 2002. Coccidiosis: This poultry disease's impact is anything but 

paltry. Healthy Animals, On-line quarterly USDA-ARS Newsletter. February 

Issue 10. 

Yamage, M.; Lillehoj, H. S.. 2002. Immunity to protozoan parasites in poultry. 

In "Modern Concepts of Immunology in Veterinary Medicine-Poultry 

Immunology," In Advances in Medical and Veterinary Virology, Immunology 

and Epidemiology, T. Matthew. Thajema ed., Thajema Press, pp183-242. 

2003 ARS Immunology Research Worskshop 169 
Min, W.; Lillehoj, H. S.; Burnside, J.; Weining, K. C.; Staeheli, P.; Nichols, M. 

2002. Adjuvant effects of IL-1beta, IL-8, IL-15, IFN-alpha, IFN-gamma, TGFbeta and Lymphotactin on DNA vaccination against Eimeria acervulina. Vaccine. 

20:267-274. 

Gehad, A. E.; Lillehoj, H. S.; Hendrick III, G.L.; Mashaly, M. 2002. I. The role 

of cytokines and hormones in the initiation of humoral immunity using Tindependent and T-dependent antigens. Develop. Comp. Immunol. 26:751-759. 

Gehad, A. E.; Lillehoj, H. S.; Hendrick, G. L.; Mashaly, M. 2002. II. The effects 

of T-independent and T-dependent antigens on the distribution of lymphocyte 

populations. Devel. Comp. Immunol. 26:761-771. 

Min, W.; Lillehoj, H. S.; Guangxing, L.; Sohn, E. J.; Miyamoto, T. 2002. 

Development and characterization of monoclonal antibodies to chicken 

interleukin-15. Vet. Immunol. Immunopathol. 88:49-56. 

Emara, M. G.; Lapierre, R.L.; Greene, G. M.; Knieriem, M.; Rosenberger, J. K.; 

Kim, C.; Lillehoj, H. S. 2002. Phenotypic variation among three broiler pure lines 

for Marek’s Disease, coccidiosis and antibody response to sheep red blood cells. 

Poultry Science 81:642-648. 

Olah, I.; Gumati, K. H.; Nagy, N.; Magyar, A.; Kaspers, B.; Lillehoj, H. S. 2002. 

Diverse expression of the K-1 antigen by cortico-medullary and reticular 

epithelial cells of the bursa of Fabricius in chicken and guinea fowl. Devel. Comp

Immunology 26:481-488. 

Emara, M. G.; Kim, H.; Lapierre, R. R.; Lakshmanan, N.; Pollock, D. L.; Sadjadi, 

M.; Lillehoj, H. S. 2002. Genetic diversity at the major histocompatibility 

complex (B) and microsatellite loci in three commercial broiler pure lines. Poul. 

Sci. 81:1609-1617. 

Babu, U.; Scott, M.; Myers, M. J.; Okamura, M.; Gaines, D.; Raybourne, R.; 

Yancy, H. F.; Lillehoj, H. S.; Heckert, R. 2002. Effects of live attenuated and 

killed Salmonella vaccine on T-lymphocyte mediated immunity in laying hens. 

Vet. Immunol. Immunopathol. 6729:1-6. 

Min, W.; Lillehoj, H. S.; Fetterer, R. 2002. Identification of an alternatively 

spliced transcript isoform of the common cytokine receptor γ chain. Biochem.

Biophys. Res. Com. 299: 321-327. 

Min, W.; Lillehoj, H. S.; 2002. Sequence and functional properties of chicken 

interleukin-17. J. Interferon & Cytokine Res. 22:1125-1130. 

Lillehoj, H. S.; Min. W. G. 2002. Gallus gallus mRNA for interleukin-16. 

Genbank. Accession Number. AJ508678. 

2003 ARS Immunology Research Worskshop 170 
Lillehoj, H. S.; Min, W. G. 2002. Gallus gallus mRNA for putative common 

cytokine receptor gamma chain β. Genbank. Accession Number. AJ419896. 

Lillehoj, H. S.; Min, W. G. 2002. Gallus gallus mRNA for putative common 

cytokine receptor gamma chain α. Genbank. Accession Number. AJ419898. 

Pinard-van der Laan, M.H.; Lillehoj, H. S.; Zhu, J. 2003. Disease resistance and 

transmission. Genetic resistance and transmission of avian parasites. In “Poultry 

Genetics, Breeding and Biotechnology”, W. M. Muir, and S. E. Aggrey eds. 

CABI Publishing, UK. 

Zhu J.; Lillehoj, H. S.; Allen, P.C.; Min, W.; Van Tassell, C.; Sonstegard, T. S.; 

Cheng, H. H.; Pollock, D. L.; Sadjadi, M; Emara, M. 2003. Mapping quantitative 

trait loci associated with resistance to coccidiosis and growth. Poultry 

Science:82:9-16. 

Dalloul, R.A.; Lillehoj, H. S.; Shellem, T.A.; Doerr, J.A. 2002. Effect of Vitamin 

A Deficiency on Host Intestinal Immune Response to Eimeria acervulina in 

Broiler Chickens. Poult. Sci. 81: 1509-1515. 

Zhou, H.; Lillehoj, H. S.; Lamont, S. 2003. Association of chicken interferon-γ

genotype and protein level with antibody response kinetics. Avian Diseases: 

46:869-876. 

Dalloul, R. A.; Lillehoj, H. S.; Shellem, T. A.; Doerr, J. A. 2003. Enhanced 

mucosal immunity against Eimeria acervulina in broilers fed a Lactobacillusbased probiotic. Poul. Sci. 82:62-66. 

Babu, U.; Scott, M.; Myers, M. J.; Okamura, M.; Gaines, D.; Raybourne, R.; 

Yancy, H. F.; Lillehoj, H. S.; Heckert, R. 2002. Effects of live attenuated and 

killed Salmonella vaccine on T-lymphocyte mediated immunity in laying hens. 

Vet. Immunol. Immunopathol. 6729:1-6. 

Xie, H.; Raybourne, R. B.; Babu, U.; Lillehoj, H. S.; Heckert, R. A. 2003. Effects 

of CPG on IL-6 and nitric oxide production, surface antigen expression, 

phagocytosis, proliferation and apoptosis of chicken macrophage cell line HD11 

cells. Dev. Comp. Immunol. 27(9):823-834. 

Okamura, M.; Lillehoj, H. S.; Raybourne, R. B.; Babu, U.; Heckert, R. 2003. 

Antigen-specific lymphocyte proliferation, Interleukin-6 (IL-6) and IL-2 

production in chickens immunized with outer membrane protein of Salmonella 

enteritidis. Avian Diseases. 47:1331-1338. 

Min, W.; Lillehoj, H. S.; Kim, S.; Zhu, J. J.; Beard, H.; Alkharouf, N.; 

Matthews, B. F. 2003. Profiling local gene expression changes associated with 

2003 ARS Immunology Research Worskshop 171 
Eimeria maxima and Eimeria acervulina using cDNA microarray. Appl Microbiol 

Biotechnol 62:392-399. 

Lillehoj, H. S.; Okamura, M. 2003. Host immunity and vaccine Development to 

coccidia and Salmonella infections in chickens. J. Poul. Sci 40:151-193. 

Withanage, G.S.; Sasai, K.; Fukata, T.; Miyamoto, T.; Lillehoj, H. S.; Baba, E. 

2003. Increased lymphocyte subpopulations and macrophages in the ovaries and 

oviducts of laying hens infected with Salmonella enterica serovar Enteritidis. 

Avian Pathol 32:583-590. 

Constantinoiu, C.C.; Lillehoj, H. S.; Matsubayashi, M.; Hosoda, Y.; Tani, H.; 

Matsuda, H.; Sasai. L,K.; Baba, E. 2003. Analysis of cross-reactivity of five new 

chicken monoclonal antibodies which recognize the apical complex of Eimeria 

using confocal laser immunofluorescence assay Vet. Parasitol. 118:29-35. 

Lillehoj, H. S.; Dalloul, R.; Min, W. 2003.Enhancement of intestinal immunity to 

coccidiosis using cytokines, oligodinucleotides and probiotics. 2003. World 

Poultry: Special Edition. pp . 18-23. 

Dalloul, R. A.; Lillehoj, H. S.; Shellem, T. A.; Doerr, J. R. 2003. Intestinal 

Immunomodulation by Vitamin A Deficiency and Lactobacillus-based Probiotic 

in Eimeria acervulina-infected Broiler Chickens. Avian Dis. 47:1313-1320. 

Jenkins, M.; Allen, P.; Fetterer, R.; Lillehoj, H. S.; Miska, K. 2003. Are there 

immunovariants of coccidia that can affect vaccine efficacy? Watt Poultry USA. 

Okamura, M.; Lillehoj, H. S.; Raybourne, R. B.; Babu, U. S.; Heckert, R. A. 

2004. Cell-mediated Immune Responses to a Killed Salmonella enteritidis 

Vaccine: Comp. Immunol Microbiol Infect Dis. 27:255-272.

Lillehoj, H. S.; Min, W.G.; Dalloul, R. A. 2004. Recent Progress on the Cytokine 

Regulation of Intestinal Immune Responses to Eimeria. Poul. Poul. Science. 

83:611-623. 

Min, W.; Lillehoj, H. S. 2004. Identification and characterization of chicken 

interleukin-16 cDNA. Dev. Comp. Immunol. 28:153-162. 

Constantinoiu, C.C.; Lillehoj, H. S.; Matsubayashi, M.; Tani, H.; Matsuda, H.; 

Sasai, L.K.; Baba, E. 2004. Characterization of stage-specific and cross-reactive 

antigens from Eimeria acervulina by chicken monoclonal antibodies. J. Vet. Med. 

Sci. 66:403-408. 

Erf, G. F.; Bersi, T. K.; Lillehoj, H. S. 2003. A role of interferon gamma in 

autoimmune vitiligo of Smyth line chickens. FEMS Immunology and Medical 

Microbiology. In press. 

2003 ARS Immunology Research Worskshop 172 
Babu, U.; Dalloul, R.A.; Okamura, M.; Lillehoj, H. S.; Xie, H.; Raybourne, R.; 

Gaines, I.; Heckert, R. A. 2004. Salmonella enteritidis Clearance and Immune 

Responses in Chickens Following Salmonella Vaccination and Challenge. 

Veterinary Immunol Immunopathol. In press. 

Miska, K.; Fetterer, R.; Min, W.; Lillehoj, H. S. Identification of a heat shock 

protein 90 gene in two Eimeria species: Expression and evolutionary analysis. 

Molecular and Biochemical Parasitology. In press. 

Lillehoj, H. S.; Guangxing, L. 2004. Nitric oxide production by macrophages 

stimulated with sporozoites, lipopolysaccharide, or IFN-γ and its dynamic 

changes in SC and TK strains of chickens infected with Eimeria tenella. .Avian 

Diseases. 48:244-253. 

Min, W.; Lillehoj, H. S.;. Ashwell, C.M.; van Tassell, C.; Matukumalli, L.K.; 

Han, J.; Lillehoj, E.P. EST analysis of Eimeria-activated intestinal intra-epithelial 

lymphocytes in chickens. Develop. Comparative Immunology. In press. 

Ding, X.; Lillehoj, H. S.; Quiroz, M.A.; Berensee, Erich; Lillehoj, Erik. 3 

Protective Immunity Against Eimeria acervulina Following In ovo Immunization 

with a Recombinant Subunit Vaccine and Cytokine Genes. In press. 

Okamura, M.; Lillehoj, H. S.; Raybourne, R.B.; Babu, U.S.; Heckert, R.A.; Sasai, 

K.; Baba, E.; Lillehoj, E.P. Differential Responses of Macrophages to Salmonella 

enterica Serovars Enteritidis and Typhimurium. Vet. Immunol. Immunopathol. In 

press. 

Dalloul, R. A.; Lillehoj, H. S.; Okamura, M.; Xie, H.; Min, W.; Ding, X.; 

Heckert, R.A. 2004. In Vivo Effects of CpG Oligodeoxynucleotide on Eimeria 

Infection in Chickens. Avian Diseases. InPress. 

Dalloul, R.; Lillehoj, H. S.; Ding, X.A.; . Klinman, D.M.C.; Heckert, R.A. In 

ovo administration of CpG oligodeoxynucleotides and the recombinant 

microneme protein MIC2 protects against Eimeria infections. In press. 

Lunney JK

Solano-Aguilar, G.I.; Vengroski, K.; Beshah, E.; Lunney, J.K. Isolation and 

Purification of lymphocyte subsets from gut associated lymphoid tissue in 

neonatal swine. J. Immunol. Methods. 2000, v. 241: p. 185-199. 

Navarro, P.; Christenson, R.; Ekhardt, G.; Bosworth, B.; Lunney, 

J.K.;Rothschild, M.; Lemke, J.; Butler, J.E. Genetic differences in the frequency 

of the hinge variants of porcine IgA is breed dependent. Vet. Immunol. 

Immunopathol. 2000 v.73:p. 287-295. 

2003 ARS Immunology Research Worskshop 173 
Alvarez, B.; Sanchez, C.; Bullido, R.; Marina, A.; Lunney, J.K.; Alonso, F.; 

Ezquerra, A.; Dominguez, J. Porcine SWC3 is a member of the SIRP family and 

associates with protein-tyrosine phosphatase SHP-1. Tissue Antigens. 2000, v. 

55:p. 342-351. 

Solano-Aguilar, G.I.; Beshah, E.; Vengroski, K.; Zarlenga, D.; Jauregui, L.; 

Cosio, M.; Douglass L.W.; Dubey, J.P.; Lunney, J.K. Cytokine and Lymphocyte 

profile after Toxoplasma gondii infection in miniature swine. Proc. 16th Cong. 

Intnl. Pig Vet. Soc.2000, Australia. p.183. 

Solano-Aguilar, G.I.; Urban, J.F.; Beshah, E.; Schopf, L.; Kringel, H.; Domingo 

,A.; Dubey, J.P.; Lunney, J.K. Mucosal immune response of young pigs after 

Toxoplasma gondii infection. Proc. 16th Cong. Intnl. Pig Vet. Soc., 2000, 

Australia. P.171. 

Johnson, P.; Keen, N.T.; Lunney, J.K.; Sadowsky, M.J. Conference Editorial: 

Agricultural Microbes Genome 2. Comparative and Functional Genomics. 2001. 

v. 2. p. 10-13. [http://www3.interscience.wiley.com/cgibin/issuetoc?ID=77002017]. 

Solano-Aguilar, G.I.; Beshah, E.; Vengroski, K.; Zarlenga, D.; Jauregui, L.; 

Cosio, M.; Douglass, L.W.; Dubey, J.P.; Lunney, J.K. Cytokine and lymphocyte 

profile after oral infection with Toxoplasma gondii oocysts. Scandinavian Journal 

of Immunology. 2001. v. 54. p. 93. 

Solano-Aguilar, G.I.; Beshah, E.; Vengroski, K.; Zarlenga, D.; Jauregui, L.; 

Cosio, M.; Douglass, L.W.; Dubey, J.P.; Lunney, J.K. Cytokine and lymphocyte 

profile in miniature swine after oral infection with Toxoplasma gondii oocysts. 

International Journal of Parasitology. 2001. v. 31. p. 187-195. 

 Solano-Aguilar, G.I.; Vengroski, K.; Beshah, E.; Douglass, L.W.; Lunney, J.K. 

Characterization of lymphocyte subsets from mucosal tissues in neonatal swine. 

Dev. Comp. Immunol. 2001, v. 25: p. 245-263. 

Jauregui, L.H.; Higgins, J.; Zarlenga, D.; Dubey, J.P.; Lunney, J.K. 

Development of a Real-Time PCR Assay for Detection of Toxoplasma gondii in 

Pig and Mouse Tissues. J. Clin. Microbiol. 2001, v. 39, :p. 2065-2071. 

Thacker, E.; Summerfield, A.; McCullough, K.; Ezquerra ,A.; Dominguez, J.; 

Alonso, F.; Lunney, J.K.; Sinkora J.; Haverson K.. Summary of workshop 

findings for porcine myelomonocytic markers. Vet. Immunol. Immunopathol. 

2001, v. 80, p.93-109. 

 Davis, W. C.; Haverson, K.; Saalmüller, A.; Yang, H.; Lunney, J.K.; Hamilton, 

M. J.; Pescovitz, M. D. Analysis of monoclonal antibodies reacting with

2003 ARS Immunology Research Worskshop 174 
molecules expressed on γδ T cells. Vet. Immunol. Immunopathol.2001, v. 80, p. 

53-62. 

Haverson,. K.; Saalmüller,. A.; Alvarez,. B.; Alonso, F.; Bailey,. M.; Bianchi, 

A.T.J.; Boersma,. W.J.A.; Chen,. Z.; Davis, W.C.; Dominguez, J.; Engelhardt, H.; 

Ezquerra, A.; Grosmaire, L.S.; Hamilton, M. J.; Hollemweguer, E.; Huang, C.; 

Khanna, K. V.; Kuebart, G., Lackovic; G., Ledbetter, J.A.; Lee, R.; Llanes, D.; 

Lunney, J.K.; McCullough, K.C.; Molitor, T.; Nielsen, .J.; Niewold, T.A.; 

Pescovitz, M.D.; Perez, de la Lastra, J.; Rehakova, Z.; Salmon, H.; Schnitzlein, 

W.M.; Seebach, J.; Simon, A.; Sinkora, J.; Sinkora, M.; Stocks, C.R.; 

Summerfield, A.; Sver, L.; Thacker , E.; Valpotic, I.; Yang, H.; Zuckermann, 

F.A.; Zwart, R. Overview of the Third International Workshop on Swine 

Leukocyte Differentiation Antigens. Vet. Immunol. Immunopathol. 2001, v. 80, 

p. 5-23. 

Lunney, J.K.; Fossum, C.; Alm, G.V.; Steinbach, F.; Wattrang, E. Veterinary 

immunology: opportunities and challenges. Trends Immunol. 2002, v. 23: p. 4-6. 

Solano-Aguilar, G.I.; Zarlenga, D.; Beshah, E.; Vengroski, K; Gasbarre, L.; 

Junker, D.; Cochran, M.; Weston, C.; Valencia, D.; Chiang, C.; Dawson H.; 

Urban J,; Lunney, J.K. . Limited effect of recombinant porcine Interleukin-12 on 

porcine lymphocytes due to a low expression of IL-122 receptor. Vet. Immunol. 

Immunopathol. 2002, v. 89, p.133-148. [Correction 95:183, 2003]. 

Solano-Aguilar, G.I.; Galina, L.; Douglass, L.W.; Lunney, J.K. Developmental 

immunity in neonatal pigs monitored by the function of peripheral blood 

mononuclear cells. Proc. 17th Cong. Intnl. Pig Vet. Soc., Iowa. 2002, vol. 1, p. 

259. 

Dawson, H.; Nishi, S.; Beshah, E.; Solano-Aguilar, G.; Zarlenga, D.; Urban, J.F.; 

Lunney, J.K. Functional genomic assessment of porcine disease resistance: real 

time assays of porcine immune gene expression. Proceedings 7th World Congress 

on Genetics Applied to Livestock Production. 2002, #13-41. 

Lunney, J.K. In Search of Disease-Resistant Pigs. National Hog Farmer, Apr 15, 

2003, pp.30-34. 

Martens, G W.; Lunney, J.K.; Baker, J.E.; Smith, D.M. Rapid Assignment of 

Swine Leukocyte Antigen (SLA) Haplotypes in Pedigreed Herds using a 

Polymerase Chain Reaction Based Assay. Immunogenetics. 2003, v. 55, p. 395-

401. 

Dawson, H.D.; Beshah, E; Nishi, S.; Solano-Aguilar, G.; Morimoto, M.; Zhao, 

A.; Madden, K.B.; Ledbetter, T.K.; Dubey, J.P.; Shea-Donohue, T.; Lunney, 

J.K.; Urban, J.F. Jr. Localized multi-gene expression patterns support an evolving 

2003 ARS Immunology Research Worskshop 175 
Th1/Th2-like paradigm in response to infections with Toxoplasma gondii and 

Ascaris suum in pigs. Infection and Immunity. In Revision. 

Lunney, J.K. Are there immune gene alleles that determine whether a pig will be

healthy? Genetics of Pig Health Symposium, M. Boggess, Ed.; National Pork 

Board Press, DesMoines, IA. 2003. p.63-72. 

Bannerman, D.; Paape, M.J.; Lunney, J.K. .Immunity: Innate. In Encyclopedia of 

Animal Science; Pond, W.G., Bell, A.W. Eds.; Marcel Dekker, Inc., New York. 

2004. In Press. 

Lunney, J.K.; Paape, M.J.; Bannerman, D. Immunity: Acquired. In Encyclopedia 

of Animal Science; Pond, W.G., Bell, A.W. Eds.; Marcel Dekker, Inc., New 

York. 2004, In Press. 

Dawson, H.D.; Royaee, A.R.; Nishi, S.; Kuhar, D.; Schnitzlein ,W.M.; 

Zuckermann, F.; Urban, J.F.; Lunney, J.K. . Identification of Key Immune 

Mediators Regulating T helper 1 Responses in Swine. Vet. Immunol. 

Immunopathol. 2004, v.100, p. 105-111. 

Gerrits, R.J.; Lunney, J.K.; Johnson, L.A.; Pursel, V.G.; Rohrer, G.A.; 

Dobrinsky, J.R. A vision for artificial insemination and genomics to improve the 

global swine population. Theriogenology. 2004, In Press. 

Royaee, A.R.; Husmann, R.; Dawson, H.D.; Calzada-Nova, G.; Schnitzlein, 

W.M.; Zuckermann, F.; Lunney, J.K. 2004. Deciphering the involvement of 

innate immune factors in the development of the host responses to PRRSV 

vaccination. Vet. Immunol. Immunopathol. In revision. 

Paape MJ

Burvenich, C.;Detilleux, J.; Paape, M.J.; Massart-Leen; A.M. Physiological and 

genetic factors that influence the cows resistance to mastitis especially during 

early lactation. Proceedings of IDF Symposium on Immunology of Ruminant 

Mammary Gland. 2000. p. 9-20. 

Diez-Fraile, A.; Tomita, G.M.; Meyer, E.; Paape, M.J.; Burvenich, C. CD62L 

expression on circulating neutrophils in relation to Escherichia coli mastitis. 

Proceedings of International Dairy Federation Symposium on Immunology of 

Ruminant Mamary Gland. 2000. p. 177-178. 

Ledbetter, T.K.; Paape, M.J.; Douglass, L.W. Cytotoxic effects of peroxynitrite, 

polymorphonuclear neutrophils, free-radical scavengers, inhibitors of 

myeloperoxidase, and inhibitors of nitric oxide synthase on bovine mammary 

secretory epithelial cells. American Journal of Veterinary Research. 2001. v. 62. 

p. 286-293. 

2003 ARS Immunology Research Worskshop 176 
Paape, M.J.; Shafer-Weaver, K.; Capuco, A.V.; Van Oostveldt, K.; Burvenich, 

C. Immune surveillance of mammary tissue by phagocytic cells. Advances in 

Experimental Medicine and Biology. 2000. v. 480. p. 259-277. 

Paape, M.J.; Duenas, M.I.; Wettemann, R.P.; Douglass, L.W. Effects of 

intramammary infection and parity on calf weaning weight and milk quality in 

beef cows. Journal of Animal Science. 2000. v. 78. p. 2508-2514. 

Paape, M.J.; Contreras, A. Limitaciones legales problematica de los recuentos 

cellulares in leche de vaca y cabra en los Estados Unidos. Ovis. 2000. v. 67. p. 13-

23. 

Peters, R.R.; Domaragiri, S.; Paape, M.J.; Douglass, L.W. Evaluation of 1.6% 

phenol as a premilking and postmilking teat dip in preventing new bovine 

intramammary infections. 2000. Journal of Dairy Science. v. 83. p. 1750-1757. 

Rainard, P.; Riollet, C.; Poutrel, B.; Paape, M.J. Phagocytosis and killing of 

Staphylococcus aureus by bovine neutrophils after priming by tumor necrosis 

factor-alpha and the des-arginine derivative of C5a. American Journal of 

Veterinary Research. 2000. v. 61. p. 951-959. 

Tomita, G.M.; Wang, Y.; Paape, M.J.; Poutrel, B.; Rainard, P. Influence of 

bispecific antibodies on the in vitro bactericidal activity of bovine neutrophils 

against Staphylococcus aureus. Journal of Dairy Science. 2000. v. 83. p. 2269-

2275. 

Van Oostveldt, K.; Tomita, G.M.; Paape, M.J.; Burvenich, C. Apoptosis of 

circulating bovine neutrophils during experimentally induced Escherichia coli and 

endotoxin mastitis. Proceedings of International Dairy Federation Symposium on 

Immunology of Ruminant Mammary Gland. 2000. p. 319-325. 

Shafer-Weaver KA

Paape, M.J.; Shafer-Weaver, K.; Capuco, A.V.; Van Oostveldt, K.; Burvenich, 

C. Immune surveillance of mammary tissue by phagocytic cells. Advances in 

Experimental Medicine and Biology. 2000. v. 480. p. 259-277. 

Solano-Aguilar GI

Solano-Aguilar, G.I.; Beshah, E.; Vengroski, K.; Zarlenga, D.; Jauregui, L.; 

Cosio, M.; Douglass, L.W.; Dubey, J.P.; Lunney, J.K. Cytokine and lymphocyte 

profile in miniature swine after oral infection with Toxoplasma gondii oocysts. 

International Journal of Parasitology. 2001. v. 31. p. 187-195. 

2003 ARS Immunology Research Worskshop 177 
Solano-Aguilar, G.I.; Beshah, E.; Vengroski, K.; Zarlenga, D.; Jauregui, L.; 

Cosio, M.; Douglass, L.W.; Dubey, J.P.; Lunney, J.K. Cytokine and lymphocyte 

profile after oral infection with Toxoplasma gondii oocysts. Scandinavian Journal 

of Immunology. 2001. v. 54. p. 93. 

Solano-Aguilar, G-I; Vengroski, K.G.; Beshah, E.; Douglass, L.W; Lunney, J.K. 

Characterization of lymphocyte subsets from mucosal tissues in neonatal swine. 

Developmental-and-comparative-immunology 2001 Apr; 25(3): 245-63. 

Solano-Aguilar, G-I.; Zarlenga, D; Beshah, E; Vengroski, K; Gasbarre, L; 

Junker, D; Cochran, M; Weston, C; Valencia, D; Chiang, C; Dawson, H; Urban, 

J-F; Lunney, J-K Limited effect of recombinant porcine interleukin-12 on porcine 

lymphocytes due to a low level of IL-12 beta2 receptor. Veterinary immunology 

and immunopathology 2002 Oct 28; 89(3-4): 133-48. 

Trout JM

Jenkins, M.C;, Trout, J.; Higgins, J.; Fayer, R. The mRNA levels for 

amyloglucosidase as measured by RT-PCR correlate with infectivity and 

fecundity of Cryptosporidium parvum oocysts for neonatal BALB/c mice. 

Proceedings of the International Conference on Emerging and Infectious

Diseases. Late Breakers. 2000. p. 9. 

Jenkins, M.C.; Trout, J.; Fayer, R. Treatment of Cryptosporidium parvum 

sporozoites with antibodies to a recombinant CP47 surface antigen inhibits in 

vitro parasite development. Proceedings of the Annual Meeting of the American 

Association of Veterinary Parasitologists. Boston, Massachusetts. 2001. p. 49. 

Jenkins, M.C.; Trout, J.; Abrahamsen, M.S.; Lancto, C.A.; Higgins, J.; Fayer, R. 

Estimating viability of Cryptosporidium parvum oocysts using reverse 

transcriptase-polymerase chain reaction (RT-PCR) directed at mRNA encoding 

amyloglucosidase. Journal of Microbial Methods. 2000. v. 43. p. 97-106. 

Jenkins, M.C.; Trout, J.; Murphy, C.; Harp, J.; Higgins, J.; Wergin, W.; Fayer, R. 

Cloning and expression of a DNA sequence encoding a 41-kilodalton 

Cryptosporidium parvum oocyst wall protein. Clinical and Diagnostic Laboratory 

Immunology. 1999. v. 6(6). p. 912-920. 

Tuo W

Goff, W.L.; Johnson, W.C.; Parish, S.M.; Barrington, G.M.; Tuo, W.; Valdez, 

R.A. The age-related immunity in cattle to Babesia bovis infection involves the 

rapid induction of IL-12, IFN-gamma and iNOS mRNA expression in the spleen. 

Parasite Immunology 2001. v. 23: p. 453-461. 

2003 ARS Immunology Research Worskshop 178 
Tuo, W.; Zhuang, D.; Knowles, D.P.; Cheevers, W.P.; Sy, M.S.; O'Rourke, K.I. 

Prp-c and Prp-Sc at the fetal-maternal interface. Journal of Biological Chemistry. 

2001. v276. p.18229-18234. 

Urban JF Jr 

Elliott, D.E.; Urban, J.F. Jr.; Argo, C.K.; Weinstock, J.V. Does the failure to 

acquire helminthic parasites predispose to Crohn's disease? The Federation of 

American Societies for Experimental Biology Journal. 2000. v. 14. p. 1848-1855. 

Finkelman, F.D.; Urban, J.F. The other side of the coin: The protective role of 

the type 2 (Th2) cytokines. Journal of Allergy and Clinical Immunology. 2001. v. 

107. p. 772-780. 

Kringel, H.; Dubey, J.P.; Beshah, E.; Heckert, R.; Urban, J.F. Effect of CpG 

oligodeoxynucleotide (ODN)-induced immune activation of porcine immunity to 

Toxoplasma gondii. American Association of Veterinary Parasitology. 2001. v. 

46. p. 15. 

Urban, J.F.; Schopf, L.; Madden, K.B.; Nobern-Trauth, N.; Finkelman, F.D. 

Nematode biology determines the requirements for expression of IL-4 receptor 

alpha chain on lymphoid and/or non-lymphoid cells that regulate intestinal 

expulsion. The Federation of American Societies for Experimental Biology 

Journal. 2001. v. 15. p. A690. 

Zarlenga DS

Hediger-Weithaler, B.M.; Heussler, V.T.; Eicher, R.; Zakher, A.; Zarlenga, D.S.;

Gasbarre, L.C.; Waldvogel, A.S. Interferon- and interleukin 4 mRNA expression 

by PBMCs from BVDV seropositive pregnant and non-pregnant cattle. Veterinary 

Immunology and Immunopathology. 2000. v. 77. p. 201-212. 

Solano-Aguilar, G.I.; Beshah, E.; Vengroski, K.; Zarlenga, D.; Jauregui, L.; 

Cosio, M.; Douglass, L.W.; Dubey, J.P.; Lunney, J.K. Cytokine and lymphocyte 

profile in miniature swine after oral infection with Toxoplasma gondii oocysts. 

International Journal of Parasitology. 2001. v. 31. p. 187-195. 

Solano-Aguilar, G.I.; Beshah, E.; Vengroski, K.; Zarlenga, D.; Jauregui, L.; 

Cosio, M.; Douglass, L.W.; Dubey, J.P.; Lunney, J.K. Cytokine and lymphocyte 

profile after oral infection with Toxoplasma gondii oocysts. Scandinavian Journal 

of Immunology. 2001. v. 54. p. 93. 

Wang, Y.; Zarlenga, D.S.; Paape, M.J.; Dahl, G.E. Recombinant bovine soluble 

CD14 sensitizes the mammary gland to lipopolysaccharide. Vet-immunolimmunopathol. Amsterdam : Elsevier. May 2002. v. 86 (1/2) p. 115-124. 

2003 ARS Immunology Research Worskshop 179 
Boston, Massachusetts 

The Jean Mayer USDA Human Nutrition Research Center on Aging 

http://hnrc.tufts.edu/

Han SN

Han, S.N.; Leka, L.S.; Lichtenstein, A.H.; Ausman, L.M.; Meydani, S.N. Effect 

of therapeutic lifestyle change (TLC) diet on immune functions of moderately 

hypercholesterolemic humans. Journal of Lipid Research Dec 2003. v. 44 (12) p. 

2304-2310. 

Chung, H.; Wu, D.; Gay, R.; Han, S.N.; Goldin, B.; Bronson, R.; Mason, J.; 

Smith, D.E.; Meydani, S.N. Effect of age on susceptibility to Azoxymethaneinduced colonic aberrant crypt foci formation in C57BL/6JNIA mice. Journals of 

Gerontology Biological Sciences and Medical Sciences May 2003. v. 58(5) p. 

B400-B405. 

Chung, H.; Wu, D.; Han, S.N.; Gay, R.; Goldin, B.; Bronson, R.E.; Mason, J.B.; 

Smith, D.E.; Meydani, S.N. Vitamin E supplementation does not alter 

Azoxymethane-induced colonic aberrant crypt foci formation in young or old 

mice. Journal of Nutrition Feb 2003 v. 133 (2) p. 528-532. 

Han, S.N.; Leka, L.S.; Lichtenstein, A.H.; Ausman, L.M.; Schaefer, E.J.; 

Meydani, S.N. Effect of hydrogenated and saturated, relative to polyunsaturated, 

fat on immune and inflammatory responses of adults with moderate 

hypercholesterolemia. Journal of Lipid Research Mar 2002. v. 43 (3) p. 445-452. 

Han, S.N.; Leka, L.S.; Lichtenstein, A.H.; Ausman, L.M.; Schaefer, E.J.; 

Meydani, S.N. Hydrogenated fat containing diets high in trans fatty acids do not 

adversely affect immune function in humans. The Federation of American 

Societies for Experimental Biology Journal. 2001. v. 15(5). Abstract p. A1094. 

Han, S.N.; Meydani, M.; Wu, D.; Bender, B.S.; Smith, D.E.; Vina, J.; Cao, G.; 

Prior, R.L.; Meydani, S.N. Effect of long-term dietary antioxidant 

supplementation on influenza virus infection. Journals of Gerontology Biological 

Sciences and Medical Sciences. 2000. v. 55A. p. B1-B8. 

Han, S.N.; Wu, D.; Ha, W.K.; Beharka, A.A.; Smith, D.; Bender, B.S.; Meydani, 

S.N. Vitamin E supplementation increases Th1 cytokine production in old mice 

infected with influenza virus infection. Immunology. 2000. v.100. p. 487-493. 

2003 ARS Immunology Research Worskshop 180 
Meydani S

Adolfsson, O.; Huber, B.T.; Meydani, S.N. Vitamin E increases IL-2 production 

by T cells from old mice through a post-transcriptional mechanism. The 

Federation of American Societies for Experimental Biology Journal. 2001. v. 

15(5). Abstract p. A293. 

Beharka, A.A.; Meydani, M.; Wu, D.; Leka, L.S.; Meydani, A.; Meydani, S.N. 

IL-6 production does not increase with age. Journals of Gerontology Biological 

Sciences and Medical Sciences. 2001. v. 56A(2). p. B81-B88. 

Beharka, A.A.; Paiva, S.; Leka, L.S.; Ribaya-Mercado, J.; Russell, R.M.; 

Meydani, S.N. Effect of age on the gastrointestinal-associated mucosal immune 

response of humans. Journals of Gerontology Biological Sciences and Medical 

Sciences. 2001. v. 56A(5). p. B218-B223. 

Bugianesi, R.; Serafini, M.; Simone, F.; Wu, D.; Meydani, S.; Ferro-Luzzi, A.; 

Azzini, E.; Maiani, G. High-performance liquid chromatography with coulometric 

electrode array detector for the determination of quercetin levels in cells of the 

immune system. Analytical Biochemistry. 2000. v. 284. p. 296-300. 

Chung, H.; Wu, D.; Gay, R.; Goldin, B.; Bronson, R.; Mason, J., Smith, D.; 

Meydani, S.N. Age mice are more susceptible to Azoxymethane (AOM)-induced 

colon cancer. The Federation of American Societies for Experimental Biology 

Journal. 2001. v. 15. Abstract p. A994. 

Han, S.N.; Leka, L.S.; Lichtenstein, A.H.; Ausman, L.M.; Schaefer, E.J.; 

Meydani, S.N. Hydrogenated fat containing diets high in trans fatty acids do not 

adversely affect immune function in humans. The Federation of American 

Societies for Experimental Biology Journal. 2001. v. 15(5). Abstract p. A1094. 

Han, S.N.; Wu, D.; Ha, W.K.; Beharka, A.A.; Smith, D.; Bender, B.S.; Meydani, 

S.N. Vitamin E supplementation increases Th1 cytokine production in old mice 

infected with influenza virus infection. Immunology. 2000. v.100. p. 487-493. 

Han, S.N.; Meydani, M.; Wu, D.; Bender, B.S.; Smith, D.E.; Vina, J.; Cao, G.; 

Prior, R.L.; Meydani, S.N. Effect of long-term dietary antioxidant 

supplementation on influenza virus infection. Journals of Gerontology Biological 

Sciences and Medical Sciences. 2000. v. 55A. p. B1-B8. 

Hayek, M.G.; Meydani, S.N. Effect of vitamin E on infection and 

immunity.Armbrecht, H.J., Coe, R.M., Morley, J.E., editors. Johns Hopkins 

University Press, Baltimore, MD. The Science of Geriatrics, 2000. v. 2. p. 583-

594. 

2003 ARS Immunology Research Worskshop 181 
King, J.C.; Black, R.E.; Doyle, M.P.; Fritsche, K.L.; Halbrook, B.H.; Levander, 

O.A.; Meydani, S.N.; Walker, W.A.; Woteki, C.E. Foodborne illnesses and 

nutritional status: A statement from an American Society for Nutritional Sciences 

working group. Journal of Nutrition. 2000. v. 130. p. 2613-2617. 

Meydani, S.N.; Fawzi, W.; Han, S.N. The effect of vitamin deficiencies (e and a) 

and supplementation on infection and immune response. Nestec, Ltd.,Vevey, 

Switzerland. Nutrition, Immunity, and Infectious Diseases in Infants and 

Children. 2000. v. 45. p. 213-241. 

Meydani, S.N.; Meydani, M. Effect of dietary supplementation with Black

currant seed oil on the immune response of healthy elderly subjects and Dietary 

conjugated linoleic acid influences the immune response of young and old 

C57BL/6NCrlBR mice. Annual Bibliography of Significant Advances in Dietary 

Supplement Research. 2000. Abstract p. 25-26. 

Meydani, S.N.; McElreavy, T.S.; Meydani, M. Antioxidants and immune

function. Fiatarone MA, editor. CRC Press, Boca Raton, FL. Exercise, Nutrition, 

and the Older Woman. 2000. p. 183-205. 

Meydani, S.N. Advances in preventive medicine: nutrition and immune function 

in the elderly. Goldstein, A.L., editor. Frontiers in Biomedicine. Kluwer 

Academic/Plenum Publishers, New York, NY. 2000. p. 19-29. 

Meydani, S.N.; Beharka, A.A. Vitamin E and immune response in the aged. 

Karger, A.G. editor. Basel, Switzerland. Bibliotheca Nutrition et Dieta, v. 55. 

2000. p. 148-158. 

Wu DY

Wu, D.; Hayek, M.G.; Meydani, S.N. Vitamin E and macrophage cyclooxygenase 

regulation in the aged. Journal of Nutrition. 2001. v. 131. p. 382S-388S. 

Wu, D.; Meydani, M.; Han, S.N.; Leka, L.S.; Meydani, S.N. Effect of dietary 

supplementation with fish oil in combination with different levels of vitamin E on 

immune response in healthy elderly human subjects. The Federation of American 

Societies for Experimental Biology Journal. 2000. v. 14. Abstract p. A238. 

East Lansing, Michigan 

The Avian Disease and Oncology Laboratory 

http://www.msu.edu/~arsadol/index.htm

2003 ARS Immunology Research Worskshop 182 
Bacon LD

Bacon, L.D.; Hunt, H.D.; Cheng, H.H. Genetic resistance to Marek's Disease. 

Hirai, K., editor. Springer Verlag, New York, NY. Marek's Disease. 2001. p. 121-

142. 

Bacon, L.D.; Hunt, H.D.; Cheng, H.H. A review of the development of chicken 

lines to resolve genes determining resistance to diseases. Poultry Science. 2000. v. 

79. p. 1082-1093. 

Witter, R.L.; Bacon, L.D.; Hunt, H.D.; Silva, R.E.; Fadly, A.M. Avian leukosis 

virus subgroup J infection profiles in broiler breeder chickens: associations with 

virus transmission to progeny. Avian Disease. 2001. 44:913-931. 

Cheng HH

Bacon, L.D.; Hunt, H.D.; Cheng, H.H. Genetic resistance to Marek's Disease. 

Hirai, K., editor. Springer Verlag, New York, NY. Marek's Disease. 2001. p. 121-

142. 

Bacon, L.D.; Hunt, H.D.; Cheng, H.H. A review of the development of chicken 

lines to resolve genes determining resistance to diseases. Poultry Science. 2000. v. 

79. p. 1082-1093. 

Liu, H. C.; Kung, H.J.; Fulton, J.E.; Morgan, R.M.; Cheng, H.H. Growth 

hormone interacts with the Marek's disease virus SORF2 protein and is associated 

with disease resistance in chicken. Proceedings of the National Academy of 

Sciences USA. 2001. v. 98. p. 9203-9208. 

Maki-Tanila, A.; Cheng, H.H.; Delany, M.E.; Burnside, J.; Mizuno, S. First 

report on chicken genes and chromosomes. 2000. Cytogenetics and Cell Genetics. 

2000. v. 90. p. 169-218. 

Schmid, M.; Nanda, I.; Guttenbach, M.; Steinlein, C.; Hoehn, H.; Schartl, M.; 

Haaf, T.; Weigend, S.; Fries, R.; Buerstedde, J.M.; Wimmers, K.; Burt, D.W.; 

Smith, J.; A'Hara, S.; Law, A.; Griffin, D.K.; Bumstead, N.; Kaufman, J.; 

Phomsom, P.A.; Burke, T.; Groenen, M.A.M.; Cheng, H.H. First report on 

chicken genes and chromosomes. Cytogenetics and Cell Genetics. 2000. v. 90. p. 

169-218. 

Suchyta, S.P.; Cheng, H.H.; Burnside, J.; Dodgson, J.B. Comparative mapping of 

chicken anchor loci orthologous to genes on human chromosomes 1, 4 and 9. 

Animal Genetics 2001. v. 32. p. 12-18. 

2003 ARS Immunology Research Worskshop 183 
Yonash, N.; Cheng, H.H.; Hillel, J.; Heller, E.D.; Cahaner, A. DNA 

microsatellites linked to quantitative trait loci affecting antibody response and 

survival rate in meat-type chickens. Poultry Science. 2001. v. 80. p. 22-28. 

Zhu, J.; Lillehoj, H.S.; Cheng, H.H.; Pollock, D.; Sadjadi, M.; Emara, M. 

Screening for highly heterozygous chickens in outbred commercial broiler lines to 

increase detection power for mapping quantitative trait loci. Poultry Science. 

2001. v. 80. p. 6-21. 

Zhu, J.; Lillehoj, H.S.; Cheng, H.H.; Pollock, D.; Emara, M.G. Screening for 

highly heterozygous chickens in outbred commercial broiler lines to increase 

detection power for mapping quantitative trait loci. Poultry Science. 2001. v. 80. 

p. 6-12. 

Zhu, J.; Lillehoj, H.; Emara, M.; Cheng, H.; Pollock, D.; Sadaji, M.; Allen, P.; 

Yun, C. Mapping QTL associated with disease resistance to coccidiosis in 

commercial broiler chickens. 89th Poultry Science Meeting. 2000. p. 9. 

Zhu, J.; Lillehoj, H.S.; Allen, P.; Cheng, H.H.; Emara, M.G.; Sadjadi, M.; 

Pollock, D. Mapping quantitative trait loci affecting disease resistance to avian 

coccidiosis. Proceedings of the 73rd Northeastern Conference on Avian Diseases. 

2001. p. 34. 

Zhu, J.; Lillehoj, H.S.; van Tassel, C.; Emara, M.; Allen, P.; Cheng, H.; Pollock, 

D.; Sadjadi, M.; Sonstegard, T. Mapping QTL loci affecting growth and disease 

resistance to avian coccidiosis. Proceedings of the International Animal 

Agricultural and Food Sciences Conference. 2001. p. 43. 

Zhu, J.; Lillehoj, H. S.; Allen, P.; Cheng, H. H.; Emara, M. G.; Sadjadi, M.; 

Pollock, D. 2001. Mapping quantitative trait loci affecting disease resistance to 

avian coccidiosis. Proc. 73rd Northeastern Conference on Avian Diseases, June 

27-29. Virginia- Maryland Regional College of Veterinary Medicine, University 

of Maryland, College Park, MD. p34. 

Zhu, J.; Lillehoj, H. S.; van Tassel, C.; Emara, M.; Allen, P.; Cheng, H.; Pollock, 

D.; Sadjadi, M.; Sonstegard, T. 2001. Mapping QTL loci affecting growth and 

disease resistance to avian coccidiosis. Proc. Int. Animal Agricultural and Food 

Sciences Conference. July 24-28, Indianapolis, IN. p43. 

Fadly AM

Fadly, A.M.; Silva, R.F.; Lee, L.F. Antigenic characterization of selected field 

isolates of subgroup J avian leukosis virus. Kaleta, E.F., Payne, L.N., HeffelsRedmann, U., editors. Proceedings, International Symposium on ALV-J and 

Other Avian Retroviruses. Rauischholzhausen, Germany. 2000. p. 13-22. 

2003 ARS Immunology Research Worskshop 184 
Fadly, A.M. Isolation and identification of avian leukosis viruses: a review. 

Avian Pathology. 2000. 29:529-535. 

Hunt, H.; Leathers, V.; Lamichhane, C.; Rosales, G.; Jensen, E.; McKay, J.; 

Fadly, A. Analysis of serum antibody to ALV-J in commercial broilers: 

correlation between ELISA, flow cytometry and Kaleta, E.F., Payne, L.N., 

Heffels-Redmann, U., editors. Proceedings, International Symposium on ALV-J 

and Other Avian Retroviruses. Rauischholzhausen, Germany. 2000. p. 127-140. 

Hunt, H.; Lupiani, B.; Fadly, A.M. Recombination between ALV-J and 

endogenous subgroup E viruses. Kaleta, E.F., Payne, L.N., Heffels-Redmann, U., 

editors. Proceedings, International Symposium on ALV-J and Other Avian 

Retroviruses. Rauischholzhausen, Germany. 2000. p. 50-60. 

Hussain, A.I.; Shanmugam, V.; Switzer, W.M.; Tsang, S.X.; Fadly, A.M; Thea, 

D.; Helfand R.; Bellini, W.; Folks, T.M.; Heneine, W. Lack of evidence of

endogenous avian leukosis virus and endogenous avian retrovirus transmission to 

measles, mumps and rubella vaccine recipients. Emerging Infectious Diseases. 

2001. 7:66-72. 

Lee, L.F.; Fadly, A.M.; Hunt, H.D. Avian leukosis virus subgroup J envelope 

gene product for diagnosis and immunogenic composition. United States Patent 

#6,146,641. 2000. 

Williams, S.M.; Reed, W.M.; Fadly, A.M. Influence of age of exposure on the 

response of line 0 and line 63 chickens to infection with subgroup J avian leukosis 

virus. Kaleta, E.F., Payne, L. N., Heffels-Redmann, U., editors. Proceedings, 

International Symposium on ALV-J and Other Avian Retroviruses. 

Rauischholzhausen, Germany. 2000. p. 67-76. 

Witter, R.L.; Bacon, L.D.; Hunt, H.D.; Silva, R.E.; Fadly, A.M. Avian leukosis 

virus subgroup J infection profiles in broiler breeder chickens: associations with 

virus transmission to progeny. Avian Disease. 2001. 44:913-931. 

Hunt HD

Bacon, L.D.; Hunt, H.D.; Cheng, H.H. A review of the development of chicken 

lines to resolve genes determining resistance to diseases. Poultry Science. 2000. v. 

79. p. 1082-1093. 

Bacon, L.D.; Hunt, H.D.; Cheng, H.H. Genetic resistance to Marek's Disease. 

Hirai, K., editor. Springer Verlag, New York, NY. Marek's Disease. 2001. p. 121-

142. 

Hunt, H.D.; Lupiani, B.; Fadly, A.M. Recombination between ALV-J and 

endogenous subgroup E viruses. Kaleta, E.F., Payne, L.N., Heffels-Redmann, U., 

2003 ARS Immunology Research Worskshop 185 
editors. Proceedings, International Symposium on ALV-J and Other Avian 

Retroviruses. Rauischholzhausen, Germany. 2000. p. 50-60. 

Hunt, H.D.; Lupiani, B.; Miller, M.M.; Gimeno, I.; Lee, L.F.; Parcells, M.S. 

Marek's disease virus down regulates surface expression of MHC (B complex) 

class I (BF) glycoproteins during active but not latent infection of chicken cells. 

Virology. 2001. v. 282. p. 198-205. 

Hunt, H.D.; Leathers, V.; Lamichhane, C.; Rosales, G.; Jensen, E.; McKay, J.; 

Fadly, A. Analysis of serum antibody to ALV-J in commercial broilers: 

correlation between ELISA, flow cytometry and . . . Kaleta, E.F., Payne, L.N., 

Heffels-Redmann, U., editors. Proceedings, International Symposium on ALV-J 

and Other Avian Retroviruses. Rauischholzhausen, Germany. 2000. p. 127-140. 

Hunt, H.D.; Lupiani, B.; Miller, M.M.; Gimeno, I.M.; Lee, L.F.; Parcells, M.S. 

Marek's disease virus down regulates surface expression of MHV (B complex) 

class I (BF) glycoproteins during active but not latent infection of chicken cells. 

Virology. 2001. v. 282(1). p. 198- 205. 

Lee, L.F.; Fadly, A.M.; Hunt, H.D. Avian leukosis virus subgroup J envelope 

gene product for diagnosis and immunogenic composition. United States Patent 

#6,146,641. 2000. 

Parcells, M.S.; Lin, S.F.; Dienglewicz, R.L.; Majerciak, V.; Robinson, D.R.; 

Chen, H.C.; Wu, Z.; Dubyak, G.R.; Brunovskis, P.; Hunt, H.D.; Lee, L.F.; Kung, 

H.J. Marek's disease virus (MDV) encodes an interleukin-8 homolog (vIL-8): 

Characterization of the vIL-8 protein and a vIL-8 deletion mutant MDV. Journal 

of Virology. 2001. v. 75(11). p. 5159-5173. 

Preisler, M.T.; Weber, P.S.D.; Tempelman, R.J.; Erskine, R.J.; Hunt, H.D.;

Burton, J.L. Glucocorticoid receptor down-regulation in neutrophils of 

periparturient cows. American Journal of Veterinary Research. 2000. v. 61. p. 14-

19. 

Preisler, M.T.; Tempelman, R.J.; Erskine, R.J.; Hunt, H.D.; Burton, J.L. 

Glucocorticoid receptor expression profiles in mononuclear leukocytes of 

periparturient Holstein cows. Journal of Dairy Science. 2000. v. 83. p. 38-47. 

Witter, R.L.; Bacon, L.D.; Hunt, H.D.; Silva, R.E.; Fadly, A.M. Avian leukosis 

virus subgroup J infection profiles in broiler breeder chickens: associations with 

virus transmission to progeny. Avian Disease. 2001. 44:913-931. 

2003 ARS Immunology Research Worskshop 186 
Lee LF

Fadly, A.M.; Silva, R.F.; Lee, L.F. Antigenic characterization of selected field 

isolates of subgroup J avian leukosis virus. Kaleta, E.F., Payne, L.N., HeffelsRedmann, U., editors. Proceedings, International Symposium on ALV-J and 

Other Avian Retroviruses. Rauischholzhausen, Germany. 2000. p. 13-22. 

Hunt, H.D.; Lupiani, B.; Miller, M.M.; Gimeno, I.M.; Lee, L.F.; Parcells, M.S. 

Marek's disease virus down regulates surface expression of MHV (B complex) 

class I (BF) glycoproteins during active but not latent infection of chicken cells. 

Virology. 2001. v. 282(1). p. 198- 205. 

Kung, H.J.; Xia, L.; Brunovskis, P.; Li, D.; Liu, J.L.; Lee, L.F. Meq: An MDVspecific bZIP transactivator with transforming properties. Hirai K, editor. 

Springer-Verlag, Berlin. Current Topics in Microbiology and Immunology. 2001. 

p. 245-260. 

Lee, L.F.; Fadly, A.M.; Hunt, H.D. Avian leukosis virus subgroup J envelope 

gene product for diagnosis and immunogenic composition. United States Patent 

#6,146,641. 2000. 

Lee, L.F.; Wu, P.; Sui, D.; Ren, D.; Kamil, J.; Kung, H.J.; Witter, R.L. The 

complete unique long sequence and the overall genomic organization of the GA 

strain of Marek's disease virus. Proceedings of the National Academy of Sciences 

USA. 2000. v. 97(11). p. 6091- 6096. 

Lupiani, B.; Lee, L.F.; Reddy, S.M. Protein-coding content of the sequence of 

Marek's disease virus. Hirai K, editor. Springer-Verlag, Berlin. Current Topics in 

Microbiology and Immunology. 2001. p. 159- 190. 

Parcells, M.S.; Lin, S.F.; Dienglewicz, R.L.; Majerciak, V.; Robinson, D.R.; 

Chen, H.C.; Wu, Z.; Dubyak, G.R.; Brunovskis, P.; Hunt, H.D.; Lee, L.F.; Kung, 

H.J. Marek's disease virus (MDV) encodes an interleukin-8 homolog (vIL-8): 

Characterization of the vIL-8 protein and a vIL-8 deletion mutant MDV. Journal 

of Virology. 2001. v. 75(11). p. 5159-5173. 

Silva, R.F.; Lee, L.F.; Kutish, G.F. The genomic structure of Marek's disease 

virus. Hirai K, editor. Springer-Verlag, Berlin. Current Topics in Microbiology 

and Immunology. 2001. p. 143-158. 

Wu, P.; Reed, W.M.; Lee, L.F. Glycoproteins H and L of Marek's disease virus 

form a hetero-oligomer essential for translocation and cell surface expression. 

Archives of Virology. 2001. v. 146. p. 983-992. 

2003 ARS Immunology Research Worskshop 187 
Reddy SM

Lupiani, B.; Lee, L.F.; Reddy, S.M. Protein-coding content of the sequence of 

Marek's disease virus. Hirai K, editor. Springer-Verlag, Berlin. Current Topics in 

Microbiology and Immunology. 2001. p. 159- 190. 

Miles, A.M.; Reddy, S.M.; Morgan, R.W. Coinfection of specific- pathogen-free 

chickens with Marek's disease virus (MDV) and chicken infectious anemia virus: 

effect of MDV pathotype. Avian Diseases. 2001. v. 45(1). p. 9-18. 

Reddy, S.M.;, Witter, R.L.; Gimeno, I.M. Development of a quantitativecompetitive polymerase chain reaction assay for serotype 1 Marek's disease virus. 

Avian Diseases. 2000. v. 44(4). p. 770-775. 

Silva RF

Fadly, A.M.; Silva, R.F.; Lee, L.F. Antigenic characterization of selected field 

isolates of subgroup J avian leukosis virus. Kaleta, E.F., Payne, L.N., HeffelsRedmann, U., editors. Proceedings, International Symposium on ALV-J and 

Other Avian Retroviruses. Rauischholzhausen, Germany. 2000. p. 13-22. 

Silva, R.F.;, Lee, L.F.; Kutish, G.F. The genomic structure of Marek's disease 

virus. Hirai K, editor. Springer-Verlag, Berlin. Current Topics in Microbiology 

and Immunology. 2001. p. 143-158. 

Witter, R.L.; Bacon, L.D.; Hunt, H.D.; Silva, R.E.; Fadly, A.M. Avian leukosis 

virus subgroup J infection profiles in broiler breeder chickens: associations with 

virus transmission to progeny. Avian Disease. 2001. 44:913-931. 

Witter RL

Lee, L.F.; Wu, P.; Sui, D.; Ren, D.; Kamil, J.; Kung, H.J.; Witter, R.L. The 

complete unique long sequence and the overall genomic organization of the GA 

strain of Marek's disease virus. Proceedings of the National Academy of Sciences 

USA. 2000. v. 97(11). p. 6091- 6096. 

Reddy, S.M.; Witter, R.L.; Gimeno, I.M. Development of a quantitativecompetitive polymerase chain reaction assay for serotype 1 Marek's disease virus. 

Avian Diseases. 2000. v. 44(4). p. 770-775. 

Witter, R.L. Part II: Marek's disease vaccines - how the virus fights back. World 

Poultry. 2001. v. 17(2). p. 38-39. 

Witter, R.L. Part 1: Marek's disease vaccines - the viral battlefield. World 

Poultry. 2001. v. 17. p. 42-44. 

2003 ARS Immunology Research Worskshop 188 
Witter, R.L.; Bacon, L.D.; Hunt, H.D.; Silva, R.E.; Fadly, A.M. Avian leukosis 

virus subgroup J infection profiles in broiler breeder chickens: associations with 

virus transmission to progeny. Avian Disease. 2001. 44:913-931. 

Witter, R.L. Protective efficacy of Marek's disease vaccines. Hirai K, editor. 

Springer-Verlag, Berlin. Current Topics in Microbiology and Immunology. 2001. 

p. 58-90. 

Witter, R.L. Determinants of early transmission of ALV-J in commercial broiler 

breeder chickens. Kaleta, E. F., Payne, L.N., Heffels-Redmann, U., editors. 

Proceedings, International Symposium on ALV-J and Other Avian Retroviruses. 

Rauischholzhausen, Germany. 2000. p. 216-225. 

Mississippi State, Mississippi 

Poultry Research Unit 

http://www.ars.usda.gov/pandp/locations/locations.htm?modecode=64-06-20-00

Branton SL

Branton, S. L. Mycoplasma vaccines in layer chickens. 45th Ann. Mgt. of State 

Contact Representatives and Inspectors. Gulfport, MS. May 15, 2001. National 

Poultry Improvement Plan. 

Branton, S. L. Making sense of sensitive Mycoplasma. 2001 Tecnica 

International. January 2001. p. 56-58. 

Branton, S.L.; Lott, B.D.; May J.D.; Maslin, W.R.; Pharr, G.T.; Bearson, S.D.; 

Collier, S.D.; Boykin, D.L. The effects of ts-11 strain Mycoplasma gallisepticum

vaccination in commercial layers on egg production and selected egg quality 

parameters. Avian Diseases. 2000. v. 44. p. 618-623. 

Stoneville, Mississippi 

Catfish Genetics Research Unit

http://www.ars.usda.gov/pandp/locations/locations.htm?modecode=64-02-35-00

Bilodeau AL

Bilodeau, A.L.; Small, B.C.; Wolters,.W.R. Pathogen loads, clearance and 

plasma cortisol response in channel catfish, Ictalurus punctatus (Rafinesque), 

following challenge with Edwardsiella ictaluri. Journal of Fish Diseases 2003 Jul; 

26(7): 433-7. 

2003 ARS Immunology Research Worskshop 189 
Quiniou SM

Barker, K.S.; Quiniou, S.M.; Wilson, M.R.; Bengten, E.; Stuge, T.B.; Warr, 

G.W.; Clem, L.W.; Miller, N.W. Telomerase expression and telomere length in 

immortal leukocyte lines from channel catfish. Developmental-and-comparativeimmunology 2000 Sep-Oct; 24(6-7): 583-95. 

Bengten, E; Quiniou, S.M.; Stuge, T.B.; Katagiri, T.; Miller, N.W.; Clem,.L.W.; 

Warr, G.W.; Wilson, M.. The IgH locus of the channel catfish, Ictalurus 

punctatus, contains multiple constant region gene sequences: different genes 

encode heavy chains of membrane and secreted IgD. Journal-of-immunology 

2002 Sep 1; 169(5): 2488-97. 

Quiniou, S.M. Physiological and Genetic Aspects of Channel Catfish Immunity: 

I. Effect of Low Temperatures Stress on Distribution of Epidermal Mucus Cells 

as a Factor in Saprolegniosis and II. Linkage and Function of Major 

Histocompatibility Genes. 2001. Thesis. 

Shen, L.; Stuge, T.B.; Zhou, H.; Khayat, M.; Barker, K.S.; Quiniou, S.M.; 

Wilson, M; Bengten, E.; Chinchar, V.G.; Clem, L.W.; Miller, N.W. Channel 

catfish cytotoxic cells: a mini-review. Developmental-and-comparativeimmunology 2002 Mar; 26(2): 141-9. 

Waldbieser GC

Chen, H.; Waldbieser, G.C.; Rice, C.D.; Elibol, B.; Wolters, W.R.; Hanson, L.A. 

Isolation and characterization of channel catfish natural resistance associated 

macrophage protein gene. Developmental-and-comparative-immunology 2002 

Jul; 26(6): 517-31. 

Yang, F.; Ventura-Holman, T.; Waldbieser, G.C.; Lobb, C.J. Structure, genomic 

organization, and phylogenetic implications of six new VH families in the channel 

catfish. Molecular Immunology 2003 Sep; 40(5): 247-60. 

Columbia, Missouri 

Animal Physiology Research Unit 

http://www.agron.missouri.edu/ars_columbia/apru.html

2003 ARS Immunology Research Worskshop 190 
Carroll JA

Carroll, J.A.; Fritsche, K.L.; Spencer, J.D.; Gaines, A.M.; Allee, G.L.; Matteri, 

R.L.; Kattesh, H.G.; Roberts, M.P.; Beausang, L.A.; Zannelli, M.E. Effect of fish 

oil supplementation on the young pig's immunological response to an endotoxic 

challenge. Journal of Animal Science. 2001. v. 79(Suppl.2):Abstract p. 34. 

Carroll, J.A.; Matteri, R.L.; Dyer, C.J.; Beausang, L.A.; Zannelli, M.E. Impact of 

environmental temperature on response of neonatal pigs to an endotoxin 

challenge. American Journal of Veterinary Research. 2001. v. 62. p. 561-566. 

Carroll, J.A.; Touchette, K.J.; Allee, G.L.; Alberts, D.; Stocco, D.M.; Welsh, 

T.H. Effect of E. Coli exposure on production of ACTH, steroidogenic acute 

regulatory (StAR) protein and cortisol in neonatal pigs. Journal of Animal 

Science. 2001. v. 79(Suppl.2):Abstract p. 20. 

Carroll, J.A.; Alberts, D.; Parzik, D.J.; Stocco, D.M.; Welsh, T.H. Influence of 

corticotropin-releasing hormone (CRH) on the expression of steroidogenic acute 

regulatory (StAR) protein in neonatal pigs derived by Caesarian section or natural 

birth. Journal of Animal Science. 2001. v. 79(Suppl.1):Abstract p. 384. 

Maddock, K.R.; Berg, E.P.; Stahl, C.A.; Linville, M.L.; Carroll, J.A. The effects 

of à-lipoic acid (LA) on performance and health of weaned neonatal pigs. Journal 

of Animal Science. 2001. v. 79(Suppl. 1):Abstract p. 398. 

Matteri, R.L.; Carroll, J.A.; Dyer, C.J. Neuroendocrine responses to stress. 

Moberg, G.P., Mench, J.A., editors. CABI Publishing, Oxon, UK. The Biology of 

Animal Stress: Basic Principles and Implications for Animal Welfare. 2000. p. 

43-76. 

Matteri, R.L.; Dyer, C.J.; Touchette, K.J.; Carroll, J.A.; Allee, G.L. Effects of 

weaning on somatotrophic gene expression and circulating levels of insulin-like 

growth factor-1 (IGF-1) and IGF-2 in pigs. Domestic Animal Endocrinology. 

2000. v. 19. p. 247-259. 

McKee, C.A.; Carroll, J.A.; Eicher, S.D.; Zannelli, M.E.; Beausang, 

L.A.Matteri, R.L. Supplemental vitamin C and beta-glucan alter growth and the 

LPS-induced immunological responses in young pigs. Journal of Animal Science. 

2001. v. 79(Suppl.2):Abstract p. 34. 

Seaman, J.S.; Safranski, T.J.; Matteri, R.L.; Carroll, J.A. Dexamethasone 

treatment and increased growth in neonatal piglets. Journal of Animal Science. 

2001. v. 79(Suppl.2):Abstract p. 46. 

2003 ARS Immunology Research Worskshop 191 
Seaman, J.S.; Berg, E.P.; Safranski, T.J.; Carroll, J.A. Effect of dexamethasone 

treatment on growth in neonatal swine. Journal of Animal Science. 2001. v. 

79(Suppl.1):Abstract p. 30. 

Strauch, T.A.; Carroll, J.A.; Abbey, C.A.; McArthur, N.H.; Durham, S.K.; Lucy, 

M.C.; Randel, R.D.; Welsh, T.H. Evaluation of somatotrophic gene expression 

and function in 3/4 Angus or Brahman steers. Journal of Animal Science. 2001. v. 

79(Suppl.2):Abstract p. 19. 

Whitley, N.C.; Barb, C.R.; Kraeling, R.R.; Barrett, J.B.; Rampacek, G.B.; 

Carroll, J.A.; Keisler, D.H. Feed intake and serum GH, LH and cortisol in gilts 

after intracerebroventricular or intravenous injection of urocortin. Domestic 

Animal Endocrinology. 2000. v. 19. p. 209-221. 

Columbia, Missouri 

Donald Danforth Plant Science Center, Plant Genetics Research Unit 

http://www.agron.missouri.edu/ars_columbia/pgru.html

Herman EM

Helm R.M.; Cockrell G.; Connaughton C.; West C.M.; Herman E.M.; Sampson 

H.A.; Bannon G.A.; Burks A.W. Mutational analysis of the IgE-binding epitopes 

of P34/Gly m Bd 30K. Journal of Allergy and Clinical Immunology 2000 105: 2 

pt 1, 378-384. 

Okinaka, Y.; Ching-Hong, Y.; Herman E.M; Kinney, A.; Keen, N. T. (2002) The 

P34 syringolide elicitor receptor interacts with a soybean photorespiration 

enzyme, NADH-dependent hydroxypyruvate reductase. Plant Molecular Microbe 

Interactions, 15 : 1213-1218 

Herman, E.M. Genetically modified soybeans and food allergies. Journal of 

Experimental Botany. 2003, 54: 386, 1317-1319. 

Herman,E.M.; Helm, R.M.; Jung, R.; Kinney, A.J. Genetic modification removes 

an immunodominant allergen from soybean. Plant Physiology 2003 May; 132(1): 

36-43. 

Herman, E.M. (2004) Allergenic responses to legume proteins. In Genomics for 

Legume Crops. Ed. C. Brummer and R. Wilson. pp. 326-348, AOCS Press. 

2003 ARS Immunology Research Worskshop 192 
Clay Center, Nebraska 

Roman L. Hruska U.S. Meat Animal Research Center 

http://www.marc.usda.gov/

Chitko-McKown CG

Heaton, M.P.; Grosse, W.M.; Kappes, S.M.; Keele, J.W.; Chitko-McKown, 

C.G.; Cundiff, L.V.; Braun, A.; Little, D.P.; Laegreid, W.W. Estimation of DNA 

sequence diversity in bovine cytokine genes. Mammalian Genome 2001 Jan; 

12(1): 32-7. 

Laegreid, W.W.; Heaton, M.P.; Keen, J.E.; Grosse, W.M.; Chitko-McKown, 

C.G.; Smith, T.P.; Keele, J.W.; Bennett, G.L.; Besser, T.E. Association of bovine 

neonatal Fc receptor alpha-chain gene (FCGRT) haplotypes with serum IgG 

concentration in newborn calves. Mammalian Genome 2002 Dec; 13(12): 704-10. 

Orient Point, New YorkPlum Island Animal Disease Center 

http://www.ars.usda.gov/plum/

Barrera J

Rodriguez, L.L.; Barrera, J.; Kramer, E.; Lubroth, J.; Brown, F.; Golde, W.T. A 

synthetic peptide containing the consensus sequence of the G-H loop region of 

foot-and-mouth disease virus type-O VP1 and a promiscuous T-helper epitope 

induces peptide-specific antibodies but fails to protect cattle against viral 

challenge. Vaccine. 2003 Sep 8; 21(25-26): 3751-6. 

Bautista EM

Bautista, E.M.; Ferman, G.S.; Golde, W.T. Induction of lymphopenia and 

inhibition of T cell function during acute infection of swine with foot and mouth 

disease virus (FMDV). Veterinary-immunology-and-immunopathology 2003 Mar 

20; 92(1-2): 61-73. 

Bautista, E.M.; Gregg, D.; Golde, W.T. Characterization and functional analysis 

of skin-derived dendritic cells from swine without a requirement for in vitro 

propagation. Veterinary-immunology-and-immunopathology 2002 Sep 25; 88(3-

4): 131-48. 

Bautista, E.M.; Faaberg, K.S.; Mickelson, D.; McGruder, E.D. Functional 

properties of the predicted helicase of porcine reproductive and respiratory 

syndrome virus. Virology. 2002 Jul 5; 298(2): 258-70. 

2003 ARS Immunology Research Worskshop 193 
Baxt B

Duque, H.; Baxt, B. Analysis of the bovine integrin subunits beta1 and beta5 as 

receptors for foot-and-mouth disease virus. American Society for Virology. 2001. 

Abstract p. 120. 

Neff, S.; Baxt, B. The Ability of Integrin Alpha V Beta 3 to Function as a 

receptor for foot-and-mouth disease virus is not dependent on the presence of 

complete subunit cytoplasmic domains. Journal of Virology. 2001. v. 75. p. 527-

532. 

Brum MC

Wu, Q.; Brum, M.C.; Caron, L.; Koster, M.; Grubman, M.J. Adenovirusmediated type I interferon expression delays and reduces disease signs in cattle 

challenged with foot-and-mouth disease virus. Journal of interferon and cytokine 

research 2003 Jul; 23(7): 359-68. 

Caron L

Caron, L.; Flores, E.F.; Weiblen, R.; Scherer, C.F.C.; Irigoyen, L.F.; Roehe, 

P.M.; Odeon, A.; Sur, J.H. Latent infection by bovine herpesvirus type-5 in 

experimentally infected rabbits: virus reactivation, shedding and recrudescence of 

neurological disease. Veterinary Microbiology Feb 4, 2002. v. 84 (4) p. 285-295. 

Golde WT

Mbow, M.L.; Gilmore, R.D. Jr; Stevenson, B.; Golde, W.T.; Piesman, J.; 

Johnson,. B.J. Borrelia burgdorferi-specific monoclonal antibodies derived from

mice primed with Lyme disease spirochete-infected Ixodes scapularis ticks. 

Hybridoma and Hybridomics 2002 Jun; 21(3): 179-82. 

Grubman MJ

Chinsangaram, J.; Koster, M.; Grubman, M.J. Inhibition of L-deleted foot- andmouth disease virus replication by interferon alpha/beta involves double-stranded 

RNA dependent protein kinase. Journal of Virology. 2001. v. 75. p. 5498-5503. 

Grubman, M J.; Chinsangaram, J.; Koster, M.; Moraes; Mauro, P. Foot and 

Mouth Disease Virus Vaccine, Provisional Patent, Docket Number: 0128.01, 

Serial Number: 60/286, 345, Date Filed: 4/26/2001. 

Mayr, G.A.; O'Donnell, V.; Chinsangaram, J.; Mason, P.W.; Grubman, M.J. 

Immune responses and protection against foot-and-mouth disease virus (FMDV) 

challenge in swine vaccinated with adenovirus-FMDV constructs. Vaccine. 2001. 

v. 19 p. 2152-2162. 

2003 ARS Immunology Research Worskshop 194 
Mason PW

Henry, T.; O'Donnell, V.; Pacheco, J.; Gregory, J.; Mason, P. Analyses of footand-mouth disease clones expressing truncated nonstructural 3A proteins reveal 

genetic plasticity. American Society for Virology. 2001. Abstract p. 205. 

Knowles, N.; Davies, P.; Henry, T.; O'Donnell, V.; Pacheco, J.; Mason, P. 

Emergence in Asia of foot-and-mouth disease viruses with altered host Range: 

characterization of alterations in the 3A protein. Journal of Virology. 2001. v. 75. 

p. 1551-1556. 

Mason, P.; Knowles, N. Emergence and re-emergence of foot-and-mouth disease 

in Asia: identification and characterization of new strains of an old enemy. United 

States Animal Health Association. 2000. p. 291-300. 

Mayr, G.A.; O'Donnell, V.; Chinsangaram, J.; Mason, P.W.; Grubman, M.J. 

Immune responses and protection against foot-and-mouth disease virus (FMDV) 

challenge in swine vaccinated with adenovirus-FMDV constructs. Vaccine. 2001. 

v. 19 p. 2152-2162. 

O'Donnell, V.; Pacheco, J.; Henry, T.; Mason, P. Attenuation of O Taiwan footand-mouth disease virus (FMDV) in bovines is not associated with an altered 

subcellular distribution of the 3A protein. American Society for Virology. 2001. 

Abstract. p. 136. 

Pacheco, J.; Mason, P. Multiple factors affect bovine attenuation of Asian FMDV 

strains. American Society for Virology. 2001. Abstract p. 179. 

Neilan JG

Sur, J.H.; Burrage, T.G.; Neilan, J.G.; Rock, D.L.; Zsak, L. African Swine Fever 

Virus Multigene Family 360 and 530 Genes Affect Virulence, Pathogenesis and 

Immune Response in Swine. American Society for Virology. 2001. Abstract p. 

84. 

Zsak, L.; Sur, J.-H.; Burrage, T.G.; Neilan, J.G.; Rock, D.L. African Swine Fever 

Virus Multigene Families 360 and 530 Genes Promote Infected Macrophage 

Survival. Miami Nature Biotechnology Winter Symposium. 2001. Abstract p. 97. 

Zsak, L.;, Lu, Z.; Burrage, T.G.;, Neilan, J.G.; Kutish, G.F.; Moore, D.M.; Rock, 

D.L. Africa Swine Fever Virus Multigene Family 360 and 530 Genes are Novel 

Macrophage Host Range Determinants. Journal of Virology. 2001. v. 75. p. 3066-

3076. 

Rock DL

2003 ARS Immunology Research Worskshop 195 
Sur, J.H., Burrage, T.G.; Neilan, J.G.; Rock, D.L.; Zsak, L. African Swine Fever 

Virus Multigene Family 360 and 530 Genes Affect Virulence, Pathogenesis and 

Immune Response in Swine. American Society for Virology. 2001. Abstract p. 

84. 

Tulman, E.R.; Rock, D.L. Novel Virulence and Host Range Genes of African 

Swine Fever Virus. Current Opinion in Microbiology. 2001. v. 4. p. 456-461. 

Zsak, L.; Sur, J. H.; Burrage, T.G.; Neilan, J.G.; Rock, D.L. African Swine Fever 

Virus Multigene Families 360 and 530 Genes Promote Infected Macrophage 

Survival. Miami Nature Biotechnology Winter Symposium. 2001. Abstract p. 97. 

Zsak, L.; Lu, Z.; Burrage, T.G.; Neilan, J.G.; Kutish, G.F.; Moore, D.M.; Rock, 

D.L. Africa Swine Fever Virus Multigene Family 360 and 530 Genes are Novel 

Macrophage Host Range Determinants. Journal of Virology. 2001. v. 75. p. 3066-

3076. 

Rodriguez LL

Flanagan, E.B.; Zamparo, J.M.; Ball, L.A.; Rodriguez, L.L.; Wertz, G.W.

Rearrangement of the genes of vesicular stomatitis virus eliminates clinical 

disease in the natural host: new strategy for vaccine development. Journal of 

Virology 2001 Jul; 75(13): 6107-14. 

Rodriguez, L.L. Emergence and re-emergence of vesicular stomatitis in the 

United States. Virus Research 2002 May 10; 85(2): 211-9. 

Rodriguez, L.L.; Barrera, J.; Kramer, E.; Lubroth, J.; Brown, F.; Golde, W.T. A 

synthetic peptide containing the consensus sequence of the G-H loop region of 

foot-and-mouth disease virus type-O VP1 and a promiscuous T-helper epitope 

induces peptide-specific antibodies but fails to protect cattle against viral 

challenge. Vaccine. 2003 Sep 8; 21(25-26): 3751-6. 

Rodriguez, L.L.; Pauszek, S.J.; Bunch,. T.A.; Schumann, .K.R. Full-length 

genome analysis of natural isolates of vesicular stomatitis virus (Indiana 1 

serotype) from North, Central and South America. Journal of General Virology 

2002 Oct; 83(Pt 10): 2475-83. 

2003 ARS Immunology Research Worskshop 196 
Scherer C

Scherer, C.F.C.; Flores, E.F.; Weiblen, R.; Caron, L.; Irigoyen, L.F.; Neves, J.P.; 

Maciel, M.N. Experimental infection of pregnant ewes with bovine viral diarrhea 

virus type-2 (BVDV-2): effects on the pregnancy and fetus. Veterinary 

Microbiology Apr 19, 2001. v. 79 (4) p. 285-299. 

Wu Q

Wu, Q.; Brum, M.C.; Caron, L.; Koster, M.; Grubman, M.J. Adenovirusmediated type I interferon expression delays and reduces disease signs in cattle 

challenged with foot-and-mouth disease virus. Journal of interferon and cytokine 

research 2003 Jul; 23(7): 359-68. 

Zsak L

Sur, J.H.; Burrage, T.G.; Neilan, J.G.; Rock, D.L.; Zsak, L. African Swine Fever 

Virus Multigene Family 360 and 530 Genes Affect Virulence, Pathogenesis and 

Immune Response in Swine. American Society for Virology. 2001. Abstract p. 

84. 

Zsak, L.; Sur, J. H.; Burrage, T.G.; Neilan, J.G.; Rock, D.L. African Swine Fever 

Virus Multigene Families 360 and 530 Genes Promote Infected Macrophage 

Survival. Miami Nature Biotechnology Winter Symposium. 2001. Abstract p. 97. 

Zsak, L.; Lu, Z.; Burrage, T.G.; Neilan, J.G.; Kutish, G.F.; Moore, D.M.; Rock, 

D.L. Africa Swine Fever Virus Multigene Family 360 and 530 Genes are Novel 

Macrophage Host Range Determinants. Journal of Virology. 2001. v. 75. p. 3066-

3076. 

College Station, Texas 

Southern Plains Agricultural Research Center 

http://sparc.usda.gov/

Crippen TL

Crippen, T.L.; Bischoff, K.M.; Lowry, V.K.; Kogut, M.H. rP33 activates 

bacterial killing by chicken peripheral blood heterophils. Journal of Food 

Protection 2003 May; 66(5): 787-92. 

Crippen, T.L.; Sheffield, C.L.; He, H.; Lowry, V.K.; Kogut, M.H. Differential 

nitric oxide production by chicken immune cells. Developmental and 

Comparative Immunology 2003 Jun; 27(6-7): 603-10. 

2003 ARS Immunology Research Worskshop 197 
Farnell, M.B.; Crippen, T.L.; He, H.; Swaggerty, C.L.; Kogut, M.H. Oxidative 

burst mediated by toll like receptors (TLR) and CD14 on avian heterophils 

stimulated with bacterial toll agonists. Developmental and Comparative 

Immunology 2003 May; 27(5): 423-9. 

Ferro PJ

Peterson, M.J.; Aguirre, R.; Ferro, P.J.; Jones, D.A.; Lawyer, T.A.; Peterson, 

M.N.; Silvy, N.J. Infectious disease survey of Rio Grande wild turkeys in the 

Edwards Plateau of Texas. Journal of Wildlife Diseases 2002 Oct; 38(4): 826-33. 

Peterson, M.J.; Ferro, P.J.; Peterson, M.N.; Sullivan, R.M.; Toole, B.E.; Silvy, 

N.J. Infectious disease survey of lesser prairie chickens in north Texas. Journal of 

Wildlife Diseases 2002 Oct; 38(4): 834-9. 

Swaggerty, C.L.; Pevzner, I.Y.; Ferro, P.J.; Crippen, T.L.; Kogut, M.H. 

Association between in vitro heterophil function and the feathering gene in 

commercial broiler chickens. Avian Pathology 2003 Oct; 32(5): 483-8. 

He H

Farnell, M.B.; He,H.; Genovese, K.; Kogut, M.H. Pharmacological analysis of 

signal transduction pathways required for oxidative burst in chicken heterophils 

stimulated by a Toll-like receptor 2 agonist. International Immunopharmacology 

2003 Nov; 3(12): 1677-84. 

He, H.; Crippen, T.L.; Farnell, M.B.; Kogut, M.H. Identification of CpG 

oligodeoxynucleotide motifs that stimulate nitric oxide and cytokine production in 

avian macrophage and peripheral blood mononuclear cells. Developmental and 

Comparative Immunology 2003 Jun; 27(6-7): 621-7. 

He, H.; Farnell, M.B.; Kogut, M.H. Inflammatory agonist stimulation and signal 

pathway of oxidative burst in neonatal chicken heterophils. Comparative 

Biochemistry and Physiology. Part-A, Molecular and Integrative Physiology 2003 

May; 135(1): 177-84. 

He, H.; Kogut, M.H. CpG-ODN-induced nitric oxide production is mediated 

through clathrin-dependent endocytosis, endosomal maturation, and activation of 

PKC, MEK1/2 and p38 MAPK, and NF-kappaB pathways in avian macrophage 

cells (HD11). Cell Signalling 2003 Oct; 15(10): 911-7. 

2003 ARS Immunology Research Worskshop 198 
Kogut MH

Kogut, M.H.. 2001. Signal transduction pathways activated by engaging 

immunoglobulin Fc receptors on chicken heterophils. Developmental and 

Comparative Immunology 25:639-646. 

Kogut, M.H.; 2002. Dynamics of a protective avian inflammatory response: the 

role of an IL-8-like cytokine in the recruitment of heterophils to the site of organ 

invasion by Salmonella enteritidis. Comparative Immunology, Microbiology, & 

Infectious Diseases. 25:159-172. 

Kogut, M.H.; Rothwell, L., Kaiser, P. 2002. Differential effects of age on 

chicken heterophil functional activation by recombinant chicken interleukin-2. 

Developmental and Comparative Immunology 26:817-830. 

Kogut, M.H.; Crippen, T.L.; Lowry, V.K.; Farnell, M. 2002. Selective 

pharmacological inhibitors reveal the role of Syk tyrosine kinase, phospholipase 

C, phosphatidylinositol 3’-kinase, and p38 mitogen-activated protein kinase in Fc 

receptor-mediated signaling of chicken heterophil degranulation. International 

Immunopharmacology 2:963-973. 

Alfano, J.C.; Petrone, V.M.; Fehervari, T.; Nava G.; Kogut, M.H.; Nisbet, D.; 

Tellez, G. 2002. Resistance to velogenic Newcastle Disease virus in Leghorn 

chickens by use of prophylactic lymphokines. Avian Diseases 46:525-534. 

Kogut, M.H.. Immunomodulation of the avian innate immune response to 

prevent bacterial infections in poultry. Feedinfo.com 1/29/02. 

Farnell, M.B.; Crippen, T.L.; Haiqi, H.; Swaggerty, C.L.; Kogut, M.H. 2003. 

Oxidative burst mediated by Toll-like receptors (TLR) and CD14 on avian 

heterophils stimulated with bacterial Toll agonists. Developmental and 

Comparative Immunology 27:423-429. 

Kogut, M.H.; Farnell, M.; Lowry, V.K. 2003. The use of selective 

pharmacological inhibitors to delineate signal transduction pathways activated 

during complement receptor-mediated degranulation in chicken heterophils. 

International Immunopharmacology 3:693-706. 

Farnell, M.B.; Kogut, M.H.; He, H. 2003. Differential activation of signaling 

transduction pathways mediating oxidative burst by chicken heterophils in

response to stimulation with lipopolysaccharide and lipoteichoic acid. 

Inflammation 27:225-231. 

Kogut, M.H.; Rothwell, L.; Kaiser, P.. 2003. Differential regulation of cytokine 

gene expression by avian heterophils during receptor-mediated phagocytosis of 

2003 ARS Immunology Research Worskshop 199 
opsonized and nonopsonized Salmonella enteritidis. Journal of Interferon and 

Cytokine Research 23:319-327. 

Farnell, M.B.; He, H.; Genovese, K.; Kogut, M.H.. 2003. Pharmacological 

analysis of signal transduction pathways required for oxidative burst in chicken 

heterophils stimulated by a Toll-like receptor-2 agonist. International 

Immunopharmacology 3:1677-1684. 

Kogut, M.H.; Rothwell, L.; Kaiser, P. 2003. Priming by recombinant chicken 

interleukin-2 induces selective expression of IL-8 and IL-18 mRNA in chicken 

heterophils during receptor-mediated phagocytosis of opsonized and 

nonopsonized Salmonella enterica serovar Enteritidis. Molecular Immunology 

40:603-610.

Kogut, M.H.; Genovese, K.J.; Lowry, V.K. Differential activation of signal 

transduction pathways mediating phagocytosis, oxidative burst, and degranulation 

by chicken heterophils in response to stimulation with opsonized Salmonella 

enteriditis. Inflammation. 2001. v. 25. p. 7-15. 

Kogut, M.H.; Genovese, K.J.; Nisbet, D.J. Signal transduction pathways 

activated by engaging immunoglobulin Fc receptors on chicken heterophils. 

Developmental and Comparative Immunology 2001, 25: 7, 639-646. 

Kogut, M.H.; Genovese, K.J.; Stanker, L.S. Immune lymphokine-mediated 

control of salmonellosis in Swine. 2001. U.S. Patent 6,221,348. 

Kogut, M.; Lowry, V.K.; Farnell, M. Selective pharmacological inhibitors reveal

the role of Syk tyrosine kinase, phospholipase C, phosphatidylinositol-3'-kinase, 

and p38 mitogen-activated protein kinase in Fc receptor-mediated signaling of 

chicken heterophil degranulation. International Immunopharmacology 2002 Jun; 

2(7): 963-73. 

Kogut, M.H.; Pishko, E.J.; Kaspers, B.; Weining, K.C. Modulation of functional 

activities of chicken heterophils by recombinant chicken IFN- gamma. Journal of 

Interferon and Cytokine Research. 2001. v. 21. p. 85- 92. 

Lowry VK

Crippen, T.L.; Sheffield, C.L.; He, H.; Lowry, V.K.; Kogut, M.H. Differential 

nitric oxide production by chicken immune cells. Developmental and 

Comparative Immunology 2003 Jun; 27(6-7): 603-10. 

Kogut, M.; Lowry, V.K.; Farnell, M. Selective pharmacological inhibitors reveal 

the role of Syk tyrosine kinase, phospholipase C, phosphatidylinositol-3'-kinase, 

and p38 mitogen-activated protein kinase in Fc receptor-mediated signaling of 

2003 ARS Immunology Research Worskshop 200 
chicken heterophil degranulation. International Immunopharmacology 2002 Jun; 

2(7): 963-73. 

Nisbet DJ

Kogut, M.H.; Genovese, K.J.; Nisbet, D.J. Signal transduction pathways 

activated by engaging immunoglobulin Fc receptors on chicken heterophils. 

Developmental and Comparative Immunology 2001, 25: 7, 639-646. 

Nava M.G.; Nisbet, D.J.; Lowry, V.K.; Tellez, I.G. Effect of prophylactic 

administration of defined probiotic on mortality and horizontal transmission by 

Salmonella gallinarum in broiler chicks. Proceedings of the Western Poultry 

Disease Conference Davis, Calif. : University of California. 2001. (50th) p. 155-

158. 

Swaggerty CL

Swaggerty, C.L.; Pevzner, I.Y.; Ferro, P.J.; Crippen, T.L.; Kogut, M.H. 

Association between in vitro heterophil function and the feathering gene in 

commercial broiler chickens. Avian Pathology 2003 Oct; 32(5): 483-8. 

Swaggerty, C.L.; Pevzner, I.Y.; Lowry, V.K.; Farnell, M.B.; Kogut, M.H. 

Functional comparison of heterophils isolated from commercial broiler chickens. 

Avian Pathology 2003 Feb; 32(1): 95-102. 

Houston, Texas 

Children's Nutrition Research Center 

http://www.bcm.tmc.edu/cnrc/index.html

Smith C.W.

Rivera, C. A.; Tcharmtchi, M. H.; Mendoza, L.; Smith; C. W. 2003. 

Endotoxemia and hepatic injury in a rodent model of hindlimb unloading. J 

Appl Physiol 95: 1656-1663. 

Robker, R.L.; Collins, R.G.; Beaudet,A.L.; Mersmann, H.J.; Smith; C. W. 2004. 

Leukocyte Migration in Adipose Tissue of Mice Null for ICAM-1 and Mac-1 

Adhesion Receptors. Obesity Research 12: June (in press). 

Li, Z.; Rivera, C.A.; Burns, A.R.; Smith; C. W. 2004. Hind 

Limb Unloading Depresses Corneal Epithelial Wound Healing in Mice. J Applied 

Physiol 97: August (in press). 

2003 ARS Immunology Research Worskshop 201 
Anderson, D.C.; Smith; C. W."Leukocyte adhesion deficiencies" In: The 

Metabolic and Molecular Bases of Inherited Disease, 8th Edition,Volume III, 

Scriver, C.R., Beaudet, A.L., Sly, W.S., Valle, D., Childs, B., Vogelstein, B. 

(eds), McGraw-Hill, New York, Chapter 188, pp.4829-4856, 2001. 

Seo, S.M.; McIntire, L.V.; Smith; C. W. 2001. Effects of IL-8, Gro-", and LTB4 

on the adhesive kinetics of LFA-1 and Mac-1 on human neutrophils. Am. J. 

Physiol.Cell.Physiol. 281:C1568-C1578. 

Prince, J.E.; Brayton, C.F.; Fossett, M.C.; Durand, J. S.; Kaplan, L.; Smith; C. 

W.; Ballantyne, C.M. 2001. The differential roles of LFA-1 and Mac-1 in host 

defense against systemic infection with Streptococcus pneumoniae1. J.Immunol. 

166:7362-7369. 

Collins, R.G.; Jung, U.; Ramirez, M.; Bullard, D.C.; Hicks, M.J.; Smith; C. W.; 

Ley, K.; Beaudet, A.L. 2001. Dermal and pulmonary inflammatory disease in Eselectin and P-selectin double-null mice is reduced in triple-selectin-null mice. 

Blood. 98:727-735. 

Mariscalco, M.M.; Vergara, W.; Mei, J.; Smith, E.; Smith; C. W. 2002. 

Mechanisms of decreased leukocyte localization in the developing host. 

Am.J.Physiol.Heart Circ.Physiol. 282:H636-H644. 

Burns, A.R.; Smith; C. W.; Walker, D. 2003. Unique structural features that 

influence neutrophil emigration into the lung. Physiol.Rev. 83:309-336. 

Chakraborty, A.; Hentzen, E.R.; Seo, S.M.; Smith; C. W. 2003. Granulocyte 

colony-stimulating factor promotes adhesion of neutrophils. Am.J.Physiol.Cell 

Physiol. 284:C103-C110. 

Logan, Utah 

Poisonous Plant Research 

http://www.ars.usda.gov/pandp/locations/locations.htm?modecode=54-28-20-00

Gardner DR

Panter, K.E.; Gardner, D.R.; James, L.F.; Stegelmeier, B.L.; Molyneux, R.J. 

2000. Natural Toxins from Poisonous Plants Affecting Reproductive Function in 

Livestock. In. Natural and Selected Synthetic Toxins: Biological Implications. 

(A.T. Tu and W. Gaffield, eds) American Chemical Society, Washington, DC. 

Pp154-172. 

2003 ARS Immunology Research Worskshop 202 
Pfister, J.A.; Panter, K.E.; Gardner, D.R.; Stegelmeier, B.L.; Molyneux, R.J.; 

Ralphs, M.H.; Lee, S.T. Alkaloids as anti-quality factors in plants on western U.S. 

rangelands. J. Range Manage. 2001. 54:447-461. 

James LF

Panter, K.E.; Gardner, D.R.; James, L.F.; Stegelmeier, B.L.; Molyneux, R.J. 

2000. Natural Toxins from Poisonous Plants Affecting Reproductive Function in 

Livestock. In. Natural and Selected Synthetic Toxins: Biological Implications. 

(A.T. Tu and W. Gaffield, eds) American Chemical Society, Washington, DC. 

Pp154-172. 

Ralphs, M.H.; Greathouse, G.; Knight, A.P.; James , L.F. Cattle preference for 

Lambert locoweed over white locoweed. J. Range Manage. 2001. 54:265-268. 

Panter KE

Panter, K.E.; Stegelmeier, B.L. Toxicoses of Food Animals. Veterinar Clinics of 

North America: Food Animal Practice 2000. 6(3):531-544. 

Panter, K.E.; Gardner, D.R.; James, L.F.; Stegelmeier, B.L.; Molyneux, R.J. 

2000. Natural Toxins from Poisonous Plants Affecting Reproductive Function in 

Livestock. In. Natural and Selected Synthetic Toxins: Biological Implications. 

(A.T. Tu and W. Gaffield, eds) American Chemical Society, Washington, DC. 

Pp154-172. 

Pfister, J.A.; Panter, K.E.; Gardner, D.R.; Stegelmeier, B.L.; Molyneux, R.J.; 

Ralphs, M.H.; Lee, S.T. Alkaloids as anti-quality factors in plants on western U.S. 

rangelands. J. Range Manage. 2001. 54:447-461. 

Pfister JA

Launchbaugh, K.; Provenza, F.D.; Pfister, J.A. Animal response to anti- quality 

factors in forages. J. Range Manage. 2001. 54:431-440. 

Pfister, J.A.; Panter, K.E.; Gardner, D.R.; Stegelmeier, B.L.; Molyneux, R.J.; 

Ralphs, M.H.; Lee, S.T. Alkaloids as anti-quality factors in plants on western U.S. 

rangelands. J. Range Manage. 2001. 54:447-461. 

Ralphs MH

Pfister, J.A.; Panter, K.E.; Gardner, D.R.; Stegelmeier, B.L.; Molyneux, R.J.; 

Ralphs, M.H.; Lee, S.T. Alkaloids as anti-quality factors in plants on western 

U.S. rangelands. J. Range Manage. 2001. 54:447-461. 

2003 ARS Immunology Research Worskshop 203 
Ralphs, M.H.; Greathouse, G.; Knight, A.P.; James, L.F.. Cattle preference for 

Lambert locoweed over white locoweed. J. Range Manage. 2001. 54:265-268. 

Stegelmeier BL

Panter, K.E.; Stegelmeier, B.L. Toxicoses of Food Animals. Veterinar Clinics of 

North America: Food Animal Practice 2000. 6(3):531-544. 

Panter, K.E.; Gardner, D.R.; James, L.F.; Stegelmeier, B.L.;Molyneux, R.J. 

2000. Natural Toxins from Poisonous Plants Affecting Reproductive Function in 

Livestock. In. Natural and Selected Synthetic Toxins: Biological Implications. 

(A.T. Tu and W. Gaffield, eds) American Chemical Society, Washington, DC. 

Pp154-172. 

Pfister, J.A.; Panter, K.E.; Gardner, D.R.; Stegelmeier, B.L.; Molyneux, R.J.; 

Ralphs, M.H.; Lee, S.T. Alkaloids as anti-quality factors in plants on western U.S. 

rangelands. J. Range Manage. 2001. 54:447-461. 

Pullman, Washington 

Animal Disease Research Unit 

http://pwa.ars.usda.gov/pullman/adru/web2.htm

Goff WL

Davidson, W.R.; Goff, W.L.; Williams, E.S.; Barker, I.K. Editors. Iowa State 

University Press, Ames, IA. Infectious Diseases of Wild Mammals, Third Edition. 

chapter 27. 2001. p. 455-466. 

Goff, W.L.; Johnson, W.C.; Parish, S.M.; Barrington, G.M.; Tuo, W.; Valdez, 

R.A. The age-related immunity in cattle to Babesia bovis infection involves the 

rapid induction of IL-12, IFN-gamma and iNOS mRNA expression in the spleen. 

Parasite Immunology 2001. v. 23: p. 453-461. 

Johnson, W.C.; Lasri, S.; Kappmeyer, L.S.; Rhalem, A.; Sahibi, H.; Knowles, 

D.P.; Goff, W.L. Factors influencing the deployment of a Babesia caballi cELISA 

in tropical areas. Proceedings of the 10th International Symposium of Veterinary 

Laboratory Diagnosticians. 2001. p. 322-323. 

Rhalem, A.; Sahibi, H.; Lasri, S.; Johnson, W.C.; Kappmeyer, L.S.; Hamidouch, 

A.; Knowles, D.P.; Goff, W.L. Validation of a competitive enzyme-linked 

immunosorbent assay for diagnosing Babesia equi infections of Moroccan origin 

and its use in determining the seroprevalence of B. equi in Morocco. Journal of 

Veterinary Diagnostic Investigations. 2001. v. 13: p. 249-251. 

2003 ARS Immunology Research Worskshop 204 
Goff, W.L.; Johnson, W.C.; Tuo, W.; Valdez, R.A.; Parish, S.M.; Barrington, 

G.M.; Davis, W.C. Age-related innate immune response in calves to Babesia 

bovis involves IL-12 induction and IL-10 modulation. Annals of the New York 

Academy of Science 2002. v. 969: p. 164-168. 

Goff, W.L.; Johnson, W.C.; Parish, S.M.; Barrington, G.M.; Elsasser T.H.; 

Davis, W.C.; Valdez, R.A. IL-4 and IL-10 inhibition of IFN-gamma- and TNFalpha-dependent nitric oxide production from bovine mononuclear phagocytes 

exposed to Babesia bovis merozoites. Veterinary Immunology and 

Immunopathology 2002. v. 84: p. 237-251. 

Boonchit, S.; Xuan, X.; Yokoyama, N.; Goff, W.L.; Wagner, G.; Igarashi, I. 

Evaluation of an enzyme-linked immunosorbent assay with recombinant rhoptryassociated protein-1 antigen against Babesia bovis for the detection of specific 

antibodies in cattle. Journal of Clinical Microbiology 2002. v. 40: p. 3771-3775. 

Goff, W.L.; McElwain, T.F.; Suarez, C.E.; Johnson, W.C.; Brown, W.C.;

Norimine, J.; Knowles, D.P. Competitive enzyme-linked immunosorbent assay 

based on a rhoptry-associated protein 1 epitope specifically identifies Babesia 

bovis-infected cattle. Clinical and Diagnostic Laboratory Immunology 2003. v. 

10: p. 38-43. 

Goff, W.L.; Johnson, W.C.; Horn, R.H.; Barrington, G.M.; Knowles, D.P. The 

innate immune response in calves to Boophilus microplus tick transmitted 

Babesia bovis involves type-1 cytokine induction and NK-like cells in the spleen. 

Parasite Immunology 2003. v. 25: p. 185-188. 

Boonchit, S.; Xuan, X.; Yokoyama, N.; Goff, W.L.; Waghela, S.D.; Wagner, G.; 

Igarashi, I. Improved enzyme-linked immunosorbent assay using C-terminal 

truncated recombinant antigens of Babesia bovis rhoptry-associated protein 1 for 

detection of specific antibodies. Journal of Clinical Microbiology 2004. v. 42: p. 

1601-1604. 

Herrmann LM

Herrmann, L. M.; Baszler, T.V.; Knowles, D.P. PrP(c) mRNA, but not PrP(Sc) 

is found in the salivary glands of scrapie-infected sheep. Biochimica et 

Biophysica Acta. 2000. v1479. p.147-154. 

Herrmann, L. M.; Davis, W.C.; Knowles, D.P.; Wardrop, K.J.; Sy, M.S.; 

Gambetti, P.; O'Rourke, K.I. Cellular prion protein is expressed on peripheral 

blood mononuclear cells but not platelets of normal and scrapie-infected sheep. 

Haematologica. 2001. v86. p.146-153. 

2003 ARS Immunology Research Worskshop 205 
Knowles DP Jr

Bradway, D.S.; Torioni de Echaide, S.; Knowles, D.P.; Hennager, S.G.; 

McElwain, T.F. Sensitivity and specificity of the complement fixation test for 

detetgion of Anaplasma marginale persistgently infected cattle. Journal 

Veterinary Diagnostic Investigation. 2001. v. 13. p.77-79. 

Brayton, K.A.; Knowles, D.P.; McGuire, T.C; Palmer, G.H. Efficient use of 

small genome to generate antigenic diversity in tick-borne ehrlichial pathogens. 

2001. Proceedings National Academy Science v.98 p.4130-4135. 

Herrmann, L. M.; Baszler, T.V.; Knowles, D.P. PrP(c) mRNA, but not PrP(Sc) is 

found in the salivary glands of scrapie-infected sheep. Biochimica et Biophysica 

Acta. 2000. v1479. p.147-154. 

Herrmann, L.M.;, Davis, W.C.; Knowles, D.P.; Wardrop, K.J.; Sy, M.S.; 

Gambetti, P.; O'Rourke, K.I. Cellular prion protein is expressed on peripheral 

blood mononuclear cells but not platelets of normal and scrapie-infected sheep. 

Haematologica. 2001. v86. p.146-153. 

Johnson, W.C.; Lasri, S.; Kappmeyer, L.S.; Rhalem, A.; Sahibi, H.; Knowles, 

D.P.; Goff, W.L. Factors influencing the deployment of a Babesia caballi cELISA 

in tropical areas. Proceedings of the 10th International Symposium of Veterinary 

Laboratory Diagnosticians. 2001. p. 322-323. 

Rhalem, A.; Sahibi, H.; Lasri, S.; Johnson, W.C.; Kappmeyer, L.S.; Hamidouch, 

A.; Knowles, D.P.; Goff, W.L. Validation of a competitive enzyme-linked 

immunosorbent assay for diagnosing Babesia equi infections of Moroccan origin 

and its use in determining the seroprevalence of B. equi in Morocco. Journal of 

Veterinary Diagnostic Investigations. 2001. v. 13: p. 249-251. 

Shoda, L.K.; Kegerreis, K.A.; Suarez, C.E.; Mwangi, W.; Knowles, D.P.; Brown, 

W.C. Inclusion of a immunoestimulatory CpG oligodeoxynucleotide sequence in 

plasmid DNA induced IL-12, TNF-a, and NO production by bovine monocytederived macrophages. J. Leukoc Biol. 2001 Jul;70(1):103-12. 

Tuo, W.; Zhuang, D.; Knowles, D.P.; Cheevers, W.P.; Sy, M.S.; O'Rourke, K.I. 

Prp-c and Prp-Sc at the fetal-maternal interface. Journal of Biological Chemistry. 

2001. v276. p.18229-18234. 

Valdez, R.A.; McGuire, T.C.; Brown, W.C.; Davis, W.C.; Knowles, D.P. Long 

term in vivo depletion of functional CD4+ T lymphocytes from calves requires 

both thymectomy and anti CD4 monoclonal antibody treatment. 2001. 

Immuology, v.102.p.426-433. 

2003 ARS Immunology Research Worskshop 206 
O’Rourke KI

Hamir, A. N.; Cutlip, R.C.; Miller, J.M.; Williams, E.S.; Stack, M.J.; Miller, 

M.W.; O'Rourke, K.I., Chaplin, M.J. Preliminary findings on the experimental 

transmission of chronic wasting disease agent of mule deer to cattle. Journal of 

Veterinary Diagnostic Investigation. 2001. v13. p.91-96. 

Herrmann, L. M.; Davis, W.C.; Knowles, D.P.; Wardrop, K.J.; Sy, M.S.; Gambetti, P.; 

O'Rourke, K.I. Cellular prion protein is expressed on peripheral blood mononuclear 

cells but not platelets of normal and scrapie-infected sheep. Haematologica. 2001. v86. 

p.146-153. 

Kim, H.; O'Rourke, K.I.; Walter, M.; Purchase, H.G.; Enck, J.; Shin, T.K. 

Immunohistochemical detection of scrapie prion proteins in clinically normal sheep in 

Pennsylvania. Journal of Veterinary Diagnostic Investigation. 2001. v13. p.89-91. 

Koo, H. C.; Park, Y.H.; Lee, B. C.; Chae, C.; O'Rourke, K.I., Baszler, T.V. 

Immunohistochemical detection of prion protein (PrP-Sc) and epidemiological study 

of BSE in Korea. Journal of Veterinary Science. 2001. v2. p.25-31. 

Raymond, G. J.; Bossers, A.; Raymond, L.D.; O'Rourke, K.I.; McHolland, L.E.; 

Bryant, III, P.K.; Miller, M.W.; Williams, E.S.; Smits, M.; Caughey, B. 2000. 

Evidence of a molecular barrier limiting susceptibility of humans, cattle and sheep to 

chronic wasting disease. EMBO Journal. 2000. v19. p.4425-4430. 

Tuo, W.; Zhuang, D.; Knowles, D.P.; Cheevers, W.P.; Sy, M.S.; O'Rourke, K.I. Prpc and Prp-Sc at the fetal-maternal interface. Journal of Biological Chemistry. 2001. 

v276. p.18229-18234. 

Suarez CE

Camacho-Nuez, M.; Munoz, M.L.; Suarez, C.E.; McGuire, T.C.; Brown, W.C.; 

Palmer G.H. Expression of polymorphic msp1b genes during acute Anaplasma 

marginale rickettsemia. Infection and Immunity.2000. 68(4):1946-52. 

Florin-Christensen, J.; Suarez, C.E.; Florin-Christensen, M.; Wainszelbaum M.; 

Brown W.C.; McElwain, T.F.; Palmer, G.H. A unique phospholipid organization 

in bovine erythrocyte membranes. In Press, Proc Natl Acad Sci U S A. 2001 Jul 

3;98(14):7736-41. 

Florin-Christensen, J.; Suarez, C. E.; Florin-Christensen, M.; Hines, S.A.; 

McElwain, T.F.; Palmer G.H. Phosphatidylcholine formation is the predominant 

lipid biosynthetic event in the hemoparasite Babesia bovis, Molecular and 

Biochemical Parasitology. 106(1):147-56, 2000. 

2003 ARS Immunology Research Worskshop 207 
Shoda, L.K.; Kegerreis, K.A.; Suarez, C.E.; Roditi, I.; Corral, R.S.; Bertot, G.M.; 

Norimine, J.; Brown, W.C. DNA from protozoal parasites Babesia bovis, 

Trypanosoma cruzi and T. brucei is mitogenic for B lymphocytes and stimulates 

macrophage expression of interleukin- 12, tumor necrosis factor alpha and nitric 

oxide. Infection and Immunity. 69(4):2162-71. 2001. 

Shoda, L.K.; Kegerreis, K.A.; Suarez, C.E.; Mwangi, W.; Knowles, D.P.; 

Brown, W.C. Inclusion of a immunoestimulatory CpG oligodeoxynucleotide 

sequence in plasmid DNA induced IL-12, TNF-a, and NO production by bovine 

monocyte-derived macrophages. J. Leukoc Biol. 2001 Jul;70(1):103-12. 

Suarez, C.E.; Florin-Christensen, M.; Hines, S.A.; Palmer, G.H.; Brown, W.C.; 

McElwain, T.F. Characterization of allelic variation in the Babesia bovis

merozoite surface antigen-1 (msa-1) gene locus and identification of a crossreactive, inhibition sensitive MSA-1 epitope. Infection and Immunity. 

68(12):6865-70, 2000. 

Valdez RA

Goff, W.L.; Johnson, W.C.; Parish, S.M.; Barrington, G.M.; Tuo, W.; Valdez, 

R.A. The age-related immunity in cattle to Babesia bovis infection involves the 

rapid induction of IL-12, IFN-gamma and iNOS mRNA expression in the spleen. 

Parasite Immunology 2001. v. 23: p. 453-461. 

Valdez, R.A.; McGuire, T.C.; Brown, W.C.; Davis, W.C.; Knowles, D.P. Long 

term in vivo depletion of functional CD4+ T lymphocytes from calves requires 

both thymectomy and anti CD4 monoclonal antibody treatment. 2001. 

Immuology, v.102.p.426-433. 

Kearneysville, West Virginia 

National Center for Cool and Cold Water Aquaculture 

http://ncccwa.ars.usda.gov/

Palti Y

Palti, Y.; Nichols, K.M.; Waller, K.I.; Parsons, J.E.; Thorgaard, G.H. Association 

between DNA polymorphisms tightly linked to MHC class II genes and IHN virus 

resistance in backcrosses of rainbow and cutthroat trout. Aquaculture Mar 15, 

2001. v. 194 (3/4) p. 283-289. 

Thorgaard, G.H.; Bailey, G.S.; Williams, D.; Buhler, D.R.; Kaattari, S.L.; Ristow, 

S.S.; Hansen, J.D.; Winton, J.R.; Bartholomew, J.L.; Nagler, J.J.; Walsh, P.J.; 

Vijayan, M.M.; Devlin, R.H.; Hardy, R.W.; Overturf, K.E.; Young, W.P; 

Robison, B.D.; Rexroad, C.; Palti, Y. Status and opportunities for genomics 

2003 ARS Immunology Research Worskshop 208 
research with rainbow trout. Comparative Biochemistry and Physiology. Part-B, 

Biochemistry and Molecular Biology 2002 Dec; 133(4): 609-46. 

Wiens GD

Wiens, G.D.; Brown, M.; Rittenberg, M.B. Repertoire shift in the humoral 

response to phosphocholine-keyhole limpet hemocyanin: VH somatic mutation in 

germinal center B cells impairs T15 Ig function. Journal of Immunology 2003 

May 15; 170(10): 5095-102. 

Wiens, G.D.; Pascho, R.; Winton, J.R. A single Ala139-to-Glu substitution in the 

Renibacterium salmoninarum virulence-associated protein p57 results in antigenic 

variation and is associated with enhanced p57 binding to chinook salmon 

leukocytes. Applied and Environmental Microbiology 2002 Aug; 68(8): 3969-77. 

Laramie, Wyoming 

Arthropod-Borne Animal Diseases Research Laboratory 

http://uwadmnweb.uwyo.edu/Usda_Ars/home.htm

Mecham JO

Mecham, J.O.; Jochim, M.M. 2000. Development of an enzyme-linked 

immunosorbent assay for the detection of antibody to epizootic hemorrhagic 

disease of deer virus. J. Vet. Diagn. Invest. 12: 142-145. 

Mecham, J.O.; Wilson, W.C.. Application of baculovirus expressed antigens in 

enzyme-linked immunosorbent assays for detection of antibody to bluetongue and 

epizootic hemorrhagic disease viruses. American Association of Veterinary 

Laboratory Diagnosticians, Hershey, Pennsylvania, 2001, pp. 20. 

Mecham, J.O. Neutralization of bluetongue virus in invertebrate cells. American 

Society for Virology, Madison, Wisconsin, 2001, Abstr. P23-1. 

Mecham, J.O.; Wilson, W.C. 2004. Antigen capture competitive enzyme-linked 

immunosorbent assays using baculovirus-expressed antigens for diagnosis of 

bluetongue virus and epizootic hemorrhagic disease virus. J. Clinical 

Microbiology 42 (2):518-523. 

White, D.M.; Mecham, J.O. Lack of detectable bluetongue virus in skin of 

seropositive cattle: Implications for vertebrate overwintering. 3rd Internatinal 

Symposium on Bluetongue, In Press. 

 

Gerry, A.C.;Mullens, B.A.; MacLachlan, N.J.; Mecham, J.O. Seasonal 

transmission of bluetongue virus by Culicoides sonorensis (Diptera: 

2003 ARS Immunology Research Worskshop 209 
Ceratopogonidae) at a southern California dairy and evaluation of vectorial 

capacity as a predictor of bluetongue virus transmission. Journal of Medical 

Entomology. 2001. v. 38, no. 2. p. 197-209. 

Mecham, J.O. Neutralization of bluetongue virus in invertebrate cells. American 

Society for Virology. 2001. Abstract P28-1. 

Mecham, J.O., et al. Report of the committee on bluetongue and bovine 

retroviruses. Proceedings of the United States Animal Health Association. 2000. 

v.104. p. 159-168. 

Meissner, J.D.; Mecham, J.O.; Wilson, W.C. Verification of bluetongue virus S9 

segment nucleotide sequences. Virus Research. 2001. (In Press). 

Wilson, W.C.; Mecham, J.O. Molecular evolution of orbiviruses. Proceedings of 

the United States Animal Health Association. 2000. v. 104. p. 169-180. 

Wilson, W.C.; Ma, H. C.; Venter, E.H.; van Djik, A.A.; Seal, B.S.; Mecham, 

J.O. Erratum to 'Phylogenetic relationships of bluetongue viruses based on gene 

S7'. Virus Research 2001. v. 73. p. 201-202. 

Wilson WC

Johnson, D.J.; Wilson,W.C.; Paul, P.S. Validation of a reverse transcriptase 

multiplex PCR test for the serotype determination of U.S. isolates of bluetongue 

virus. Veterinary Microbiology. 2000. v. 76. p. 105-115. 

Meissner, J.D.; Mecham, J.O.;, Wilson, W.C. Verification of bluetongue virus S9 

segment nucleotide sequences. Virus Research. 2001. (In Press). 

Wilson, W.C.; Ma, H. C.; Venter, E.H.; van Djik, A.A.; Seal, B.S.; Mecham, J.O. 

Erratum to 'Phylogenetic relationships of bluetongue viruses based on gene S7'.

Virus Research 2001. v. 73. p. 201-202. 

Wilson, W.C.; Mecham, J.O. Molecular evolution of orbiviruses. Proceedings of 

the United States Animal Health Association. 2000. v. 104. p. 169-180. 

2003 ARS Immunology Research Worskshop 210 
APPENDIX

• List of Participants 

• Author Index 

2003 ARS Immunology Research Worskshop 211 
List of Participatnts 

Uma S. Babu 

Food and Drug Administration 

8301 Muirkirk Road 

Laurel, MD 20708 

301-827-8515 

301-594-0517 

usb@cfsan.fda.gov 

Cynthia Baldwin 

University of Massachusetts 

410 Paige Laboratory 

Amherst, MA 01003 

413-545-3167 

413-545-6326 

cbaldwin@vasci.umass.edu 

Rick Barb 

USDA, ARS, SAA, Animal Physiology 

Research Unit 

Russell Research Center 

P.O. Box 5677 

Athens, GA 30604-5677 

706-583-8276 

706-542-0399 

rbarb@saa.ars.usda.gov 

José Barrera 

USDA, ARS, Plum Island Animal Disease

Ctr 

P. O. Box 848 

Greenport, NY 11944-0848 

631-323-3266 

631-323-3262 

Elida M. Bautista 

USDA, ARS, Plum Island Animal Disease

Ctr 

P. O. Box 848 

Greenport, NY 11944-0848 

631-323-3292 

631-323-3295 

ebautista@piadc.ars.usda.gov 

Jay A. Berzofsky 

Center for Cancer Research, National 

Cancer Institute 

National Institute of Health 

Building 10, Room B12 

Bethesda, MD 20892 

301-496-6874 

307-496-9956 

berzofsky@helix.nih.gov 

Joseph T. Bielitzki 

Defense Advanced Research Projects 

Agency/DSO 

3701 North Fairfax Drive 

Arlington, VA 22203-1714 

703-696-5278 

jbielitzki@darpa.mil 

Lanie Bilodeau 

USDA, ARS, Catfish Genetics Research 

Unit 

141 Experiment Station Road 

Stoneville, MS 38776 

662-686-3591 

662-686-3567 

abilodeau@ars.usda.gov

2003 ARS Immunology Research Worskshop 212 
Willi K. Born 

National Jewish Medical & Research 

Center 

1400 Jackson Street 

Denver, CO 80220 

303-398-1087 

303-398-1396 

birn@ujc.org 

Sonia de Avila Botton 

USDA, ARS, Plum Island Animal 

Disease Center 

31 Clearwater Road 

Old Saybrook, CT 06475 

631-323-3325 

631-323-3006 

sbotton@piadc.ars.usda.gov 

David A. Brake 

Pfizer Animal Health R&D 

Pfizer Global Research and 

Development, Pfizer, Inc. 

M.S. 81180C2 

Groton, CT 06340 

860-441-8230 

860-715-7936 

david_brake@groton.pfizer.com

Deborah L. Brennan 

USDA, ARS, MSA 

P. O. Box 225 

141 Experiment Station Road 

Stoneville, MS 38776 

662-686-5266 

662-686-5459 

dbrennan@ars.usda.gov 

Robert E. Briggs 

USDA, ARS, National Animal Disease 

Center 

2300 Dayton Avenue 

Ames, IA 50010 

515-663-7762 

515-663-7458 

bbriggs@nadc.ars.usda.gov

Kim A. Brogden 

USDA, ARS, National Animal Disease 

Center 

P. O. Box 70, 2300 Dayton Avenue 

Ames, IA 50010 

515-663-7980 

515-663-7458 

kbrogden@nadc.ars.usda.gov 

Wendy C. Brown 

Washington State University 

Department of Veterinary Microbiology 

and Pathology 

Pullman, WA 99164-7040 

509-335-6067 

509-335-8529 

wbrown@vetmed.wsu.edu 

Mario Celso Sporotto Brum 

USDA, ARS, Plum Island Animal 

Disease Center 

Foot and Mouth Disease Unit 

P. O. Box 848 

Greenport, NY 11922 

631-323-3320 

631-323-3006 

mbrum@piadc.ars.usda.gov 

John Butler 

Department of Microbiology 

The University of Iowa 

51 Newton Road, 3550 Bowen Science 

Building 

Iowa City, IA 52242 

319-335-7776 

john-butler@uiowa.edu 

Luizinho Caron 

USDA, ARS, PIADC, UNICAMP-IBPPGBM 

1146 Boston Post Road 

Old Saybrook, CT 06475 

631-323-3320 

631-323-3355 

lcaron@piadc.ars.usda.gov

2003 ARS Immunology Research Worskshop 213 
Carol G. Chitko-McKown 

USDA, ARS, U. S. Meat Animal 

Research Center 

P. O. Box 166, State Spur 18D 

Clay Center, NE 68933-0166 

402-762-4372 

402-762-4375 

mckown@email.marc.usda.gov 

Michelle M. Colby 

Office of Science and Technology 

Policy 

438 Eisenhower Executive Office 

Building 

Washington, DC 20502 

202-456-6067 

202-456-6027 

mcolby@ostp.eop.gov 

Darrell F. Cole 

USDA, ARS, MWA 

1815 N. University Street 

Peoria, IL 61604 

309-681-6600 

309-681-6684 

dfc@mwa.ars.usda.gov 

Stephanie D. Collier 

USDA, ARS, Poultry Research Unit 

P. O. Box 5367 

Mississippi State, MS 39762 

662-320-7483 

662-320-7589 

scollier@msa-msstate.ars.usda.gov

Tawni Crippen 

USDA, ARS, SPARC 

2881 F&B Road 

College Station, TX 77845 

979-260-3773 

979-260-9332 

Rami A. Dalloul 

USDA, ARS, ANRI, Animal Parasitic 

Diseases Laboratory 

Bldg 1043, Room 101, BARC-East 

Beltsville, MD 20705 

301-504-8771 

301-504-5103 

rdalloul@anri.barc.usda.gov 

Bernadette Davillier 

USDA, ARS, NPS, APPVS 

GWCC 

5601 Sunnyside Avenue, Bldg 4-2170D 

Beltsville, MD 20705-5138 

301-504-4698 

301-504-5467 

Mari Ellen Davis 

University of Arkansas 

Department of Animal Science, AFLS

B-107 

Fayetteville, AR 72701 

479-575-5036 

479-575-5756 

Henry Dawson 

USDA, ARS, Nutrient Requirements and 

Functions Laboratory 

10300 Baltimore Avenue 

Bldg 307, Room 206, BARC-East 

Beltsville, MD 20705 

301-504-9412 ext. 278 

301-504-9062 

dawson@307.bhnrc.usda.gov 

Annie De Groot 

EpiVax, Inc. 

16 Bassett Street 

Providence, RI 02903 

401-272-2123 

401-272-7562 

Anne_DeGroot@Brown.edu

2003 ARS Immunology Research Worskshop 214 
Ann M. Donoghue 

USDA, ARS, Poultry Production and 

Product Safety Research 

POSC 303 University of Arkansas 

Fayetteville, AR 72701 

479-575-2413 

479-575-4202 

donoghue@uark.edu 

Susan Eicher 

USDA, ARS, Livestock Behavior 

Research Unit 

125 S. Russell Street, 216 Poultry, 

Purdue University 

Lafayette, IN 47907 

765-496-3665 

765-496-1993 

spruiett@purdue.edu 

Theodore Elsasser 

USDA, ARS, ANRI, Growth Biology 

Laboratory 

Bldg 20, Room 201, BARC-East 

Beltsville, MD 20705 

301-504-8281 

301-504-8623 

elsasser@anri.barc.usda.gov 

Gisela F. Erf 

University of Arkansas 

Department of Poultry 

Fayetteville, AR 72701 

479-575-8664 

479-575-7139 

gferf@uark.edu 

Mark Estes

University of Texas Medical Branch 

2-330F Children's Hospital - Department 

of Pediatrics 

Galveston, TX 77555-0732 

409-772-0434 

409-772-0460 

dmestes@utamb.edu 

Matthew J. Fenton 

University of Maryland School of 

Medicine 

685 W. Baltimore, MSTF-800 

Baltimore, MD 21201-1192 

410-706-0159 

410-706-8162 

mfenton@medicine.umaryland.edu 

Brad Fenwick 

USDA, CSREES 

800 9th Street, SW

Waterfront Center 

Washington, DC 20024 

202-401-5022 

202-401-2653 

bfenwick@csrees@usda.gov 

Geoffrey Ferman 

USDA, ARS, Plum Island Animal 

Diseases Center 

P. O. Box 848 

Greenport, NY 11944-0848 

631-323-3285 

631-323-3295 

gferman@piadc.ars.usda.gov 

Pamela J. Ferro 

USDA, ARS, SPARC 

2881 F & B Road, Building 1 

College Station, TX 77845 

979-260-9525 

979-260-9332 

ferro@ffsru.tamu.edu 

Janet Fulton 

Hy-Line International 

P. O. Box 310, 240th Street 

Dallas Center, IA 50063 

515-992-4173 

jfulton@hyline.com

2003 ARS Immunology Research Worskshop 215 
Louis Gasbarre 

USDA, ARS, Bovine Functional 

Genomics Laboratory 

Bldg. 200, BARC-East, Room 6 

Beltsville, MD 20705 

301-504-8509, 301-504-6426 

lgasbarr@anri.barc.usda.gov 

Cyril Gay 

USDA, ARS, NPS, APPVS 

GWCC 

5601 Sunnyside Avenue, Bldg 4-2194 

Beltsville, MD 20705-5138 

301-504-4786 

301-504-5467 

cgg@ars.usda.gov 

Jesse P. Goff 

USDA, ARS, National Animal Disease 

Center 

2300 Dayton Avenue 

Ames, IA 50010 

515-663-7547 

515-663-7669 

jgoff@nadc.ars.usda.gov 

Will L. Goff

USDA, ARS, Animal Disease Research 

Unit 

3003 ADBF, Washington State 

University 

Pullman, WA 99164-6630 

509-335-6003 

509-335-8328 

wgoff@vetmed.wsu.edu 

William T. Golde 

USDA, ARS, Plum Island Animal 

Diseases Center 

P. O. Box 848 

Greenport, NY 11944-0848 

631-323-3249 

631-323-3005 

wgolde@piadc.ars.usda.gov 

Ronald D. Green 

USDA, ARS, NPS, APPVS 

GWCC, 5601 Sunnyside Avenue, Bldg 

4-2164 

Beltsville, MD 20705-5138 

301-504-4736 

rdg@ars.usda.gov 

Adam Grow

USDA, APHIS, VS, NCAHP, EST

4700 River Road, Unit 43 

Riverdale, MD 20737 

301-736-3752 

adam.g.grow@aphis.usda.gov 

Andrew Hammond 

USDA, ARS, SAA 

P. O. Box 5677 

950 College Station Road 

Athens, GA 30604 

706-546-3311 

706-546-3398 

ACH@saa.ars.usda.gov 

James A. Harp 

USDA, ARS, National Animal Disease 

Center 

2300 Dayton Avenue 

Ames, IA 50010 

515-663-7533 

515-663-7669 

jharp@nadc.ars.usda.gov 

Gary J. Hausman 

USDA, ARS, Russell Research Center 

Animal Physiology Research Unit 

950 College Station Road 

Athens, GA 30605-2720 

706-583-8275 

706-542-0399 

ghausman@saa.ars.usda.gov 

2003 ARS Immunology Research Worskshop 216 
Haiqi He 

USDA, ARS 

2881 F&B Road 

College Station, TX 77845 

979-260-3771 

979-260-9332 

he@ffsru.tamu.edu 

Robert Heckert 

USDA, ARS, NPS, APPVS 

GWCC 

5601 Sunnyside Avenue, Bldg 4-2176 

Beltsville, MD 20705-5138 

301-504-5774 

301-504-5467 

rah@ars.usda.gov 

Eliot Herman 

USDA, ARS 

Plant Genetic Research Unit, Donald 

Danforth Plant Science Center 

975 N. Warson Road 

St. Louis, MO 63105 

314-587-1292 

314-587-1392 

eherman@danforthcenter.org 

Lynn M. Herrmann 

USDA, ARS, Animal Disease Research 

Unit 

Washington State University 

ADBF, Room 3003 

Pullman, WA 991640-6630 

509-335-6068 

509-335-8328 

lherrman@vetmed.wsu.edu 

Adrianna D. Hewings 

USDA, ARS, MWA 

1815 N. University Street 

Peoria, IL 61604 

309-681-6602 

309-681-6684 

Hewingsa@mwa.ars.usda.gov 

Ronald L. Horst 

USDA, ARS, National Animal Disease 

Center 

2300 Dayton Avenue 

Ames, IA 50010 

515-663-7312 

515-663-7669 

rhorst@nadc.ars.usda.gov 

Geraldine Huff 

USDA, ARS, Poultry Production and 

Product Safety Research 

POSC 303 University of Arkansas 

Fayetteville, AR 72701 

479-575-7966 

479-575-4202 

grhuff@uark.edu 

Henry D. Hunt 

USDA, ARS, Avian Diseases and 

Oncology Research 

3606 E. Mount Hope Road 

East Lansing, MI 48823-5338 

517-337-6834 

517-337-6776 

hunthd@msu.edu 

Peter J. Johnson 

USDA, CSREES 

800 9th Street, S.W.

Washington, DC 20024 

202-401-1896 

pjohnson@csrees.usda.gov 

Pete Kaiser

Institute for Animal Health 

Compton, Berkshire RG20 7NN, UK 

+44 1635 577277 

pete.kaiser@bbsrc.ac.uk 

2003 ARS Immunology Research Worskshop 217 
Ildy M. Katona 

Uniformed Services University of the 

Health Sciences 

Department of Pediatrics at USUHS 

4301 Jones Bridge Road, Room A3007 

Bethesda, MD 20814 

301-295-3365 

301-295-3898 

dthomas@usuhs.mil 

Marcus Kehrli 

USDA, ARS, National Animal Disease 

Ctr 

Virus and Prion Diseases of Livestock 

2300 Dayton Avenue 

Ames, IA 50010 

515-663-7254 

mkehrli@nadc.ars.usda.gov 

Kayoko Kimura 

USDA, ARS, National Animal Disease 

Center 

2300 Dayton Avenue 

Ames, IA 50010 

515-663-7419 

515-663-7669 

Kirk C. Klasing 

University of California 

Department of Animal Science 

1 Shields Avenue 

Davis, CA 95616 

530-752-1901 

530-752-0175 

kcklasing@ucdavis.edu 

Donald P. Knowles 

USDA, ARS, Animal Disease Research 

Unit 

Washington State University 

ADBF, Room 3003 

Pullman, WA 99164-6630 

509-335-6022 

509-335-8328 

dknowles@vetmed.wsu.edu 

Michael H. Kogut 

USDA, ARS, Southern Plains 

Agricultural Research Center 

Food and Feed Safety Research 

2881 F&B Road 

College Station, TX 77845-9594 

979-260-3772 

979-260-9332 

kogut@ffsru.tamu.edu 

John Korslund 

USDA, APHIS, VS, AHP, NCAAP, 

EST 

4700 River Road 

Riverdale, MD 20737 

301-734-5914 

301-734-7984 

john.a.korslund@aphis.usda.gov 

Tim R. Kramer 

USDA, ARS 

900 S. Shackleford Road 

Suite 509 

Little Rock, AR 72211 

501-954-8102 

501-954-9596 

tkramer@spa.ars.usda.gov 

Arthur M. Kreig 

Coley Pharmaceutical Group 

93 Worcester Street, Suite 101 

Wellesley, MA 02481 

781-431-9000 ext.1284 

781-431-0951 

akreig@coleypharma.com 

William W. Laegreid 

USDA, ARS, U. S. Meat Animal 

Research Center 

P. O. Box 166, State Spur 18D 

Clay Center, NE 68933-0166 

402-762-4177 

402-762-4375 

laegreid@email.marc.usda.gov 

2003 ARS Immunology Research Worskshop 218 
Kelly M. Lager 

USDA, ARS, National Animal Disease 

Center 

P. O. Box 70 

Ames, IA 50010 

515-63-7371 

515-663-7458 

klager@nadc.ars.usda.gov 

Jean Larson 

USDA, ARS, NAL, Animal Welfare 

Information Center 

10301 Baltimore Avenue 

Beltsville, MD 20705 

301-504-5215 

301-504-7122 

jlarson@nal.usda.gov 

Beth Lautner 

National Pork Board 

1776 NW 114th Street 

Clive, IA 50325 

515-223-2623 

515-223-2646 

beth.lautner@porkboard.org 

Robert Li

USDA, ARS, Catfish Genetics Research 

Unit 

141 Experiment Station Road 

Stoneville, MS 38776 

662-686-3535 

662-686-3567 

rli@ars.usda.gov 

Hyun Lillehoj

USDA, ARS, ANRI, APDL 

Building 1040, BARC-East 

Beltsville, MD 20705 

301-504-8771 

301-504-5103 

hlilleho@anri.barc.usda.gov 

James Lindsay 

USDA, ARS, NPS, APPVS 

GWCC 

5601 Sunnyside Avenue, Bldg 4-2182 

Beltsville, MD 20705-5138 

301-504-4674 

301-504-5467 

jal@ars.usda.gov 

John D. Lippolis 

USDA, ARS, National Animal Disease 

Center 

2300 Dayton Avenue 

Ames, IA 50010 

515-663-7446 

515-663-7669 

Linda Logan-Henfrey 

Pathobiology College of Veterinary 

Medicine 

Mailstop 1961, Texas A&M University 

College Station, TX 77843-1255 

979-845-2660 

979-862-3128 

llogan@cvm.tamu.edu 

Virginia K. Lowry 

Texas A&M University 

USDA, ARS, SPARC 

2881 F & B Road 

College Station, TX 77845 

979-260-9263 

979-260-9332 

lowry@ffsru.tamu.edu 

Joan K. Lunney 

USDA, ARS, Animal Parasitic Diseases 

Laboratory 

10300 Baltimore Avenue 

Bldg 1040, Rom 107, BARC-East 

Beltsville, MD 20705 

301-504-9368 

301-504-5306 

jlunney@anri.barc.usda.gov 

2003 ARS Immunology Research Worskshop 219 
Lolita Mangra 

USDA, ARS, NPS, APPVS 

GWCC 

5601 Sunnyside Avenue, Bldg 4-2172A 

Beltsville, MD 20705-5138 

301-504-4778 

301-504-5467 

lsm@ars.usda.gov 

Wilda Martinez

USDA, ARS, NAA 

600 East Mermaid Lane 

Wyndmoor, PA 19038 

215-233-6593 

215-233-6719 

wilda.martinez@ars.usda.gov 

Charles V. Maxwell 

University of Arkansas 

Department of Animal Science, AFLS

B-106 

Fayetteville, AR 72701 

479-575-2111 

479-575-5756 

cmaxwell@uark.edu 

Scott McVey

Pfizer Animal Health 

601 W. Cornhusker Highway 

Lincoln, NE 68527 

402-441-2817 

402-441-2760 

d.scott.mcvey@pfizer.com 

James O. Mecham 

USDA, ARS, Arthropod-Borne Animal

Diseases Research Laboratory 

P. O. Box 3965, University Station 

Laramie, WY 82071 

307-766-3620 

307-766-3500 

jmecham@uwyo.edu 

Jeremy Merchant-Forde 

USDA, ARS, LBRU 

Poultry Science Building, Purdue 

University 

125 S. Russell street 

West Lafayette, IN 47907 

705-494-6358 

705-496-1993 

merchant@perdue.edu 

Simin Meydani 

USDA, HNRC at Tufts University 

711 Washington Street 

Boston, MA 02111 

617-556-3129 

617-556-3224 

simin.meydani@tufts.edu 

Wongi Min 

USDA, ARS, Animal Parasitic Diseases

Lab. 

Animal and Natural Resources Institute 

Building 1040, BARC-East 

Beltsville, MD 20705 

301-504-8771 

301-504-5103 

wongimin@anri.barc.usda.gov 

Brian Murphy 

National Institutes of Health (NIH) 

National Institute of Allergies and 

Infectious Diseases (NIAID) 

50 South Drive MSC 8007 

Bethesda, MD 20892 

301-594-1616 

301-480-5033 

bm25f@nih.gov 

Michael Murtaugh 

Department of Veterinary PathoBiology 

University of Minnesota 

1971 Commonwealth Avenue 

St. Paul, MN 55108 

612-625-6735 

612-625-5203 

murta001@umn.edu

2003 ARS Immunology Research Worskshop 220 
Peter L. Nara 

Biological Mimetics, Inc. 

124 Byte Drive 

Frederick, MD 21702 

301-620-7691 

301-620-1827 

John Neill 

USDA, ARS, National Animal Disease 

Center 

2300 Dayton Avenue 

Ames, IA 50010 

515-663-7730 

515-663-7458 

jneill@nadc.ars.usda.gov 

Brian J. Nonnecke 

USDA, ARS, National Animal Disease 

Center 

2300 Dayton Avenue 

Ames, IA 50010 

515-663-7311 

515663-7669 

Evelyn A. Nystrom 

USDA, ARS, National Animal Disease 

Ctr. 

P. O. Box 70 

Ames, IA 50010 

515-663-7376 

515--663-7458 

enystrom@nadc.ars.usda.gov 

Yniv Palti 

USDA, ARS, National Center for Cool 

and Cold Water Aquaculture

11876 Leetown Road 

Kearneysville, WV 25430 

304-724-8340 

304-725-0351 

ypalti@ncccwa.ars.usda.gov 

Muquarrab A. Qureshi 

USDA, CSREES 

1400 Independence Avenue S.W.

Washington, DC 20250-2220 

202-401-4895 

202-401-1602 

mqureshi@csrees.usda.gov 

Narayan Rath 

USDA, ARS, Poultry Production and 

Product Safety Research 

POSC 303 University of Arkansas 

Fayetteville, AR 72701 

479-575-6189 

479-575-4202 

nrath@uark.edu 

Richard B. Raybourne

Food and Drug Administration 

8301 Muirkirk Road 

Laurel, MD 20708 

301-827-8099 

301-594-0517 

Richard.Raybourne@cfsan.fda.gov 

Timothy A. Reinhardt 

USDA, ARS, National Animal Disease 

Center 

2300 Dayton Avenue 

Ames, IA 50010 

515-663-7540 

515-663-7669 

treinhar@nadc.ars.usda.gov 

Juergen A. Richt 

USDA, ARS, National Animal Disease 

Center 

2300 Dayton Avenue 

Ames, IA 50010 

515-663-7366 

515-663-7548 

jricht@nadc.ars.usda.gov 

2003 ARS Immunology Research Worskshop 221 
Julia Ridpath 

USDA, ARS, National Animal Disease 

Center 

P.O. Box 70, 2300 Dayton Avenue 

Ames, IA 50010 

515-663-7586 

515-663-7458 

jridpath@nadc.ars.usda.gov 

Jane Robens 

USDA, ARS, NPS, APPVS 

GWCC 

5601 Sunnyside Avenue, Bldg 4-2184 

Beltsville, MD 20705-5138 

301-504-5381 

301-504-5467 

jfr@ars.usda.gov 

M. Fernanda Rodriguez 

USDA, ARS, National Center for Cool 

and Cold Water Aquaculture

11876 Leetown Road 

Kearneysville, WV 25430 

301-724-8340 ext 2184 

frodriguez@ncccwa.ars.usda.gov 

Gary A. Rohrer 

USDA, ARS, U.S. Meat Animal 

Research Center 

Spur 18D, P. O. Box 166 

Clay Center, NE 68933-0166 

402-762-4365 

402-762-4390 

rohrer@email.marc.usda.gov 

Ronald Rosenberg 

USDA, ARS, NPS, APPVS 

GWCC 

5601 Sunnyside Avenue, Bldg 4-2178 

Beltsville, MD 20705-5138 

301-504-5771 

301-504-5467 

rmr@ars.usda.gov 

James A. Roth 

Iowa State University 

VMPM, College of Veterinary Medicine 

Ames, IA 50011 

515-294-8459 

515-294-8259 

jaroth@iastate.edu 

Randy E. Sacco 

USDA, ARS, National Animal Disease 

Center 

2300 Dayton Avenue 

 Ames, IA 50010 

515-663-7354 

515-663-7458 

rsacco@nadc.ars.usda.gov 

Charles Fernando Capinos Scherer 

USDA, ARS, PIADC, University of 

Missouri 

97 High Street, Route 81 

Clinton, CT 06413-1733 

860-669-0137 

csherer@piadc.ars.usda.gov 

Ronald D. Schultz

University of Wisconsin-Madison 

Department of Pathobiological Sciences, 

School of Veterinary Medicine 

2015 Linden Drive 

Madison, WI 53706-1102 

608-263-9888 

608-262-1739 

manningj@svm.vetmed.wisc.edu 

Richard Shelby 

USDA, ARS, Aquatic Animal Health 

Laboratory 

P. O. Box 952 

Auburn, AL 36830 

334-887-3741 

334-887-2983 

2003 ARS Immunology Research Worskshop 222 
Franklin A. Sher 

National Institutes of Health, National 

Institute of Allergy and Infectious 

Diseases 

50 South Drive, MSC-8002, Building. 

50, Room 6140, 

Bethesda, MD 20892 

301-496-3535 

301-402-0890 

asher@niaid.nih.gov

C. Wayne Smith 

USDA, ARS, Baylor College of 

Medicine 

Immunology & Medicine-Thrombosis 

Research, Head, Section of Leukocyte 

Biology 

1100 Bates, Room 6014 

Houston, TX 77030-2600 

713-798-0311 

713-798-0337 

cwsmith@bcm.tmc.edu 

Douglas Smith 

Baylor University Medical Center 

3500 Gaston Avenue 

Dallas, TX 75246 

214-820-2119 

214-820-6384 

dsmith@baylorhealth.edu

Lewis Smith 

USDA, ARS, NPS, APPVS 

GWCC 

5601 Sunnyside Avenue, Bldg 4-2168 

Beltsville, MD 20705-5138 

301-504-5925 

301-504-5467 

lws@ars.usda.gov 

Joseph T. Spence 

USDA, ARS, NPS, APPVS 

GWCC 

5601 Sunnyside Avenue, Bldg 4-2186 

Beltsville, MD 20705-5138 

301-504-7050 

301-504-4725 

jts@ars.usda.gov 

Phillip J. Sprino 

Colorado Serum Company 

4950 York Street 

Denver, CO 80216-0428 

303-295-7527 

303-295-1923 

psprino@colorado-serum.com

Judith R. Stabel 

USDA, ARS, National Animal Disease 

Center 

2300 Dayton Road 

Ames, IA 50010 

515-663-7304 

515-663-7458 

jstabel@nadc.ars.usda.gov 

Charles Stephensen 

USDA, ARS, Western Human Nutrition 

Research Center 

Nutrition Department 

1 Shields Avenue, University of 

California 

Davis, CA 95616 

530-754-9266 

530-752-8966 

cstephensen@ucdavis.edu 

Donice Stewart 

USDA, ARS, NPS, APPVS 

GWCC 

5601 Sunnyside Avenue, Bldg 4-2174 

Beltsville, MD 20705-5138 

301-504-4583 

301-504-5467 

dcs@ars.usda.gov 

2003 ARS Immunology Research Worskshop 223 
M. Suresh 

Department of Pathobiological Sciences 

School of Veterinary Medicine, 

University of Wisconsin-Madison 

2015 Linden Drive 

Madison, WI 53706-1102 

608-265-9791 

608-263-0438 

sureshm@svm.vetmed.wisc.edu 

Christina Swaggerty 

USDA, ARS, SPARC, FFSRU 

2881 F & B Road 

College Station, TX 77845 

979-260-9397 

979-260-9332 

swaggerty@ffsru.tamu.edu 

David E. Swayne 

USDA, ARS, Southeast Poultry 

Research Laboratory 

RBRRC, 950 College Station Road 

Athens, GA 32604 

706-546-3433 

706-546-3161 

dswayne@seprl.usda.gov 

Wenbin Tuo 

USDA, ARS, APDL, ANRI 

Bldg 1040, Room 1, BARC-East 

Beltsville, MD 20705 

301-504-8258 

wtuo@anri.barc.usda.gov 

Joseph F. Urban 

USDA, ARS, Nutrient Requirements and 

Functions Laboratory 

10300 Baltimore Avenue 

Bldg 307-C, Room 214, BARC-East 

Beltsville, MD 20705 

301-504-5528 ext. 267 

301-504-9062 

urban@307.bhnrc.usda.gov 

Ray Waters 

USDA, ARS, National Animal Disease 

Center 

2300 Dayton Avenue 

Ames, IA 50010 

515-663-7756 

515-663-7458 

rwaters@nadc.ars.usda.gov 

David White 

USDA, ARS, ABADRL

College of Agriculture 

Department 3965, 1000 E. University 

Avenue 

Laramie, WY 82071 

307-766-3601 

307-766-3500 

dmwhite@uwyo.edu 

Ann L. Wiegers

USDA, APHIS, VS 

510 South 17th Street, Suite 104 

Ames, IA 50010 

515-232-5785 ext.503 

515-232-7120 

ann.l.wiegers@aphis.usda.gov 

Gregory Wiens

USDA, ARS, National Center for Cool 

and Cold Water Aquaculture

11876 Leetown Road 

Kearneysville, WV 25430 

304-724-8340 

304-725-0351 

gwiens@ncccwa.ars.usda.gov 

David Woodland 

Trudeau Institute 

154 Algonquin Avenue 

Soranac Lake, NY 12983 

518-891-3080 

518-891-5126 

dwoodland@trudeauinstitute.org 

2003 ARS Immunology Research Worskshop 224 
Dayong Wu 

USDA, ARS, Human Nutrition Research 

Center on Aging at Tufts University 

711 Washington Street 

Boston, MA 02111 

617-556-3368 

617-556-3224 

dayong.wu@tufts.edu 

Lijuan Yuan 

The Ohio State University 

FAHRP/OARDC 1680 Madison Avenue 

Wooster, OH 44691 

330-263-3740 

330-264-3677 

yuan.21@osu.edu 

Xiaoping Zhu 

University of Maryland at College Park 

Lab of Immunology, VA-MD Regional 

College of Veterinary Medicine 

8075 Greenmead Drive 

College Park, MD 20742 

301-314-6814 

301-314-6855 

xz60@umail.umd.edu 

Federico A. Zuckerman

University of Illinois 

College of Veterinary Medicine 

2001 South Lincoln Avenue 

Urbana, IL 61802 

217-333-7767 

217-244-7421 

fazaaa@uiuc.edu 

Kurt Zuelko 

USDA, ARS, ANRI, Biotechnology & 

Germplasm Laboratory 

Bldg 200, Room 124, BARC-East 

Beltsville, MD 20705 

301-504-5161 

301-504-5123 

kzuelke@anri.barc.usda.gov 

2003 ARS Immunology Research Worskshop 225 
Author Index 

Abdel-Azim, GA................................. 75 

Adolfsson O ........................................ 70 

Allen PC............................................ 159 

Andersen AA .................................... 132 

Araujo R.............................................. 36 

Araujo, R............................................. 76 

Arns, CW ............................................ 80 

Augustine PC .................................... 160 

Babu, Uma .......................................... 43 

Bacon LD.......................................... 183 

Badger TM........................................ 119 

Baldwin, C. L...................................... 39 

Balog JM..................................... 70, 115 

Bannantine JP...................................... 66 

Bannerman DD ........................... 55, 160 

Barrera J...................................... 47, 193 

Bautista EM .......................... 20, 79, 193 

Bautista, Elida M. ............................... 43 

Baxt B ............................................... 194 

Belisle S .............................................. 69 

Beshah E ....................................... 30, 65 

Beshah, E ............................................ 74 

Bilodeau AL................................ 75, 189 

Bono JL............................................... 55 

Born, Willi .......................................... 22 

Botton SA............................................ 79 

Boyd P................................................. 23 

Boyd, P................................................ 65 

Branton SL........................................ 189 

Bricker BJ......................................... 133 

Briggs RE.......................................... 133 

Brockmeier SL............................ 56, 134 

Brogden KA...................................... 135 

Brown, Wendy C. ............................... 23 

Brum MC ........................ 20, 79, 80, 194 

Brum, Mario C.................................... 43 

Burton, JL ........................................... 75 

Butte, N............................................... 73 

Call J................................................... 30 

Call, J.................................................. 74 

Caron L ....................................... 80, 194 

Carroll JA.......................................... 191 

Casey TA ............................................ 80 

Champagne ET.................................. 157 

Cheevers, W.P..................................... 53 

Cheng HH ................................... 27, 183 

Cheng HW .................................. 44, 132 

Cheng, H. ............................................ 54 

Chitko-McKown CG................... 55, 193 

Chung S............................................. 158 

Collier SD ........................................... 47 

Collins, R ............................................ 73 

Crippen TL.................................. 43, 197 

Dalloul R....................................... 19, 68 

Dalloul, Rami...................................... 43 

Davis, W.C.......................................... 49 

Dawson H.................. 30, 34, 60, 65, 161 

Dawson, H........................................... 74 

Dawson, H........................................... 68 

De Groot, Annie.................................. 27 

Dean-Nystrom EA ...................... 39, 143 

DeCunha, Cristina W.......................... 48 

Dellagostin, O.A. ................................ 48 

Donoghue AM .................................. 116 

Dubey JP............................... 23, 60, 162 

Dubey, JB............................................ 65 

Eiben, L............................................... 54 

Eicher SD.............................. 18, 44, 131 

Eicher, S.D.......................................... 45 

Eiting, K.T. ......................................... 55 

Elsasser TH....................................... 162 

Emara, M............................................. 77 

Estes, D. Mark..................................... 25 

Evans JD ........................................... 163 

Evans JJ............................................. 109 

Fach, SJ............................................... 49 

Fadly AM.......................................... 184 

Fayer R.............................................. 163 

Ferman GS .......................................... 20 

Ferman, Douglas Gregg...................... 43 

Ferman, Geoffrey S............................. 43 

Ferro PJ........................... 37, 71, 78, 198 

Ferro, PJ.............................................. 76 

Fetterer RH.................................. 23, 163 

Fetterer, R. .......................................... 65 

Foote, M.R. ......................................... 52 

2003 ARS Immunology Research Worskshop 226 
Foote, MR........................................... 73 

Fowler, M.A........................................ 52 

Fowler, MA......................................... 73 

Fox JM ................................................ 55 

Frank GH .......................................... 135 

Freeman, AE ....................................... 75 

Freytag T............................................. 74 

Gardner DR....................................... 202 

Gasbarre LC.......................... 23, 36, 163 

Gasbarre, LC....................................... 76 

Gasbarre. L.......................................... 65 

Gast RK............................................. 124 

Gause, W............................................. 63 

Gay RT.......................................... 32, 69 

Gay, R.T.............................................. 73 

Goff JP .......................................... 50, 58 

Goff WL...................................... 45, 204 

Goff, J. P. ............................................ 25 

Goff, J.P. ............................................. 50 

Golde WT............ 20, 47, 62, 79, 80, 194 

Golde, William T. ............................... 43 

Gregg D......................................... 20, 62 

Grosse WM......................................... 55 

Grubman MJ ................... 20, 79, 80, 194 

Grubman, Marvin................................ 43 

Hakkak R........................................... 120 

Han SN............................ 32, 69, 70, 180 

Han, SN............................................... 73 

Hansen, J.D. ........................................ 54 

Hare W................................................ 30 

Hare, W............................................... 74 

Harlan, J.M.......................................... 55 

Harp JA....................................... 50, 136 

He H...................................... 41, 43, 198 

Heaton MP .......................................... 55 

Heckert, Robert A. .............................. 43 

Herman EM....................................... 192 

Herrmann LM ............................. 28, 205 

Herrmann, L.M. .................................. 53 

Hines, SA............................................ 48 

Holt PS........................................ 50, 125 

Horst RL............................................ 138 

Horton R.............................................. 84 

Hovatter K........................................... 53 

Huff GR ...................................... 70, 116 

Huff WE...................................... 70, 117 

Hunt HD...................................... 27, 185 

Hunt, H................................................ 54 

Hwang DH ........................................ 120 

Jacob RA........................................... 121 

James A. Harp..................................... 80 

James LF ........................................... 203 

Janutenaite, J....................................... 56 

Jenkins MC ....................................... 164 

Jiang X ................................................ 74 

Johnson WC........................................ 45 

Johnson, T. A. ..................................... 45 

Johnson, T.R. ...................................... 45 

Kaiser P............................................... 45 

Kaiser, P.............................................. 76 

Kapczynski DR..................... 81, 84, 126 

Kappmeyer LS .................................... 48 

Keen JE............................................... 55 

Kehrli ME Jr ......................... 58, 75, 138 

Kelley DS.......................................... 121 

Kelm, SC............................................. 75 

Khalifeh MS........................................ 64 

Kim, JK............................................... 81 

Kimura K .................................... 58, 138 

Kimura, K. .................................... 25, 50 

King DJ....................................... 84, 126 

Klasing, Kirk C. .................................. 31 

Klesius PH ........................................ 110 

Klinman, Dennis M............................. 44 

Knowles DP Jr ................ 28, 45, 48, 206 

Knowles, D.P. ..................................... 53 

Kogut MH..... 37, 41, 43, 45, 71, 78, 199 

Kogut, MH.......................................... 76 

Koster MJ............................................ 79 

Kuck, AL............................................. 75 

Laegreid WW...................................... 55 

Lager KM.................................... 56, 139 

Ledbetter T.......................................... 30 

Ledbetter, T......................................... 74 

Lee CW............................................... 83 

Lee LF............................................... 187 

Lee, Jai Wei ........................................ 46 

Lehmkuhl HD ................................... 140 

Lillehoj HS.... 19, 37, 40, 68, 77, 81, 165 

Lillehoj, HS................................... 77, 82 

Lillehoj, Hyun S.................................. 43 

Lim CE.............................................. 112 

2003 ARS Immunology Research Worskshop 227 
Lippolis JD.......................................... 58 

Lopez, A.M. ........................................ 48 

Lowry VK................................... 71, 200 

Lunney JK................................... 60, 173 

Madden, K........................................... 63 

Maleki SJ .................................... 72, 158 

Marko M ............................................. 69 

Mason PW......................................... 195 

Mathews B.......................................... 65 

Matthews BF....................................... 37 

Matthews, BF...................................... 77 

Maue A................................................ 82 

McGuire, Travis C. ............................. 48 

McMunn KA....................................... 18 

McMunn, K.A..................................... 45 

Mecham JO................................. 51, 209 

Mengeling WL.................................. 140 

Mersmann, H....................................... 73 

Meydani S................. 32, 67, 69, 70, 181 

Meydani, SN ....................................... 73 

Miller JM .................................... 66, 141 

Miller LC ............................................ 55 

Miller, B.L. ......................................... 52 

Miller, BL ........................................... 73 

Min W............................... 37, 40, 77, 81 

Min, HS............................................... 77 

Min, W.......................................... 77, 82 

Moraes MP.......................................... 80 

Morash, M........................................... 54 

Morimoto M........................................ 65 

Morimoto, M....................................... 63 

Murtaugh, Michael.............................. 35 

Neilan JG .......................................... 195 

Neill JD................................. 59, 62, 142 

Nielsen NC........................................ 132 

Nisbet DJ........................................... 201 

Nishi SM............................................. 60 

Noakes, M........................................... 54 

Nonnecke BJ............. 25, 33, 52, 85, 142 

Nonnecke, BJ...................................... 73 

Nonnecker, B.J.................................... 50 

O’Rourke KI ..................................... 207 

Oesper JC............................................ 80 

Olsen SC ..................................... 82, 144 

Ordovas J ............................................ 70 

Paape MJ..................................... 46, 176 

Palmer MV................ 49, 66, 82, 85, 145 

Palti Y ................................... 53, 54, 208 

Panter KE.......................................... 203 

Perdue ML ........................................ 127 

Pesch B................................................ 56 

Pevzner, IY ......................................... 76 

Pfister JA........................................... 203 

Phillips, R............................................ 54 

Prior RL ............................................ 120 

Quiniou SM....................................... 190 

Rahner TE........................................... 50 

Ralphs MH........................................ 203 

Rath NC ................................ 60, 70, 118 

Raybourne, Richard ............................ 44 

Reddy SM ......................................... 188 

Register KB....................................... 145 

Reinhardt TA .............................. 58, 145 

Rexroad C III ...................................... 53 

Richt JA ............................................ 146 

Ridpath JF............................. 59, 62, 147 

Ristow, S.S.......................................... 54 

Robker, R............................................ 73 

Rock DL............................................ 196 

Rodriguez LL........................ 47, 62, 196 

Rothwell L .......................................... 45 

Rutherford, Mark ................................ 35 

Sacco RE............................... 49, 59, 148 

Scherer C..................................... 62, 197 

Schnell, S ............................................ 75 

Schoene N..................................... 30, 65 

Schoene, N.......................................... 74 

Schultz-Cherry S................................. 84 

Seal BS.............................................. 128 

Shafer-Weaver KA............................ 177 

Shea-Donohue T. .................... 30, 63, 65 

Shea-Donohue, T ................................ 74 

Shelby R.A........................................ 112 

Shoemaker CA.................................. 113 

Silva RF ............................................ 188 

Small B................................................ 75 

Smith CW.................................... 73, 201 

Solano-Aguilar GI................. 30, 65, 177 

Solano-Aguilar, G............................... 74 

Sonstegard T ....................................... 36 

Sonstegard, T ...................................... 76 

Stabel JR ............................... 64, 66, 150 

2003 ARS Immunology Research Worskshop 228 
Stabel TJ...................................... 56, 150 

Staeheli, P ........................................... 82 

Stanton TB........................................ 151 

Stegelmeier BL ................................. 204 

Stephensen CB............................ 74, 122 

Stoffegren WS..................................... 82 

Suarez CE.......................................... 207 

Suarez DL ............................. 83, 84, 129 

Suresh, M. ........................................... 41 

Swaggerty CL ....................... 37, 78, 201 

Swaggerty, CL .................................... 76 

Swayne DE.................................. 84, 130 

Tabatabai LB..................................... 151 

Thorgaard, G.H. .................................. 54 

Trout JM............................................ 178 

Tumpey T.............................. 81, 84, 131 

Tuo W ............................................... 178 

Tuo, wenbin ........................................ 65 

Tuo, Wenbin ....................................... 23 

Urban JF Jr................ 30, 60, 63, 65, 179 

Urban, J............................................... 74 

Valdez RA......................................... 208 

Van Tassel, C...................................... 76 

Van Tassell C...................................... 36 

Waldbieser GC.................................. 190 

Waters WR.... 33, 49, 52, 66, 82, 85, 151 

Waters, WR......................................... 73 

Welch T............................................... 53 

Wesley RD........................................ 155 

Whipple DL ................................ 85, 156 

Wiens GD.................................... 53, 209 

Wilson WC........................................ 210 

Winn, R.K. .......................................... 55 

Witter RL .......................................... 188 

Wolters W........................................... 75 

Woodland, David L............................. 22 

Wu DY.................................. 32, 67, 182 

Wu Q................................................. 197 

Wu, D.................................................. 73 

Xiao, Zhengguo................................... 35 

Xie, Hang ............................................ 43 

Xu D.................................................. 115 

Yersin AG......................................... 156 

Zarlenga DS .......................... 63, 65, 179 

Zhang H. ............................................. 27 

Zhang, H. ............................................ 54 

Zhao, A. .............................................. 63 

Zhao, Xin ............................................ 46 

Zimmerman, A.................................... 54 

Zsak L ............................................... 197 

Zuerner RL........................................ 156 

Zunino SJ.......................................... 123 

2003 ARS Immunology Research Worskshop 229