U. S. DEPARTMENT OF HEALTH AND HUMAN SERVICES 

National Institutes of Health 

National Institute of Allergy and Infectious Diseases 

NIAID Biodefense Research Agenda 

for CDC Category A Agents 

February 2002 

BIODEFENSE 
NIAID Biodefense Research Agenda 

for CDC Category A Agents 

February 2002 

U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES 

National Institutes of Health 

National Institute of Allergy and Infectious Diseases 

NIH Publication No. 03-5308 

February 2002 

http://biodefense.niaid.nih.gov 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

TABLE OF CONTENTS


v PREFACE 


1 INTRODUCTION 


2 AREAS OF RESEARCH EMPHASIS 


5 GENERAL RECOMMENDATIONS 


8 ANTHRAX 


14 SMALLPOX 


20 PLAGUE 


24 BOTULISM 


28 TULAREMIA 


32 VIRAL HEMORRHAGIC FEVERS 


A-1 APPENDIX A 


iii 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

PREFACE


On February 4 and 5, 2002, the National Institute of Allergy and Infectious 

Diseases (NIAID) convened the Blue Ribbon Panel on Bioterrorism and Its 

Implications for Biomedical Research. This panel of experts was brought together 

by NIAID to provide objective expertise on the Institute’s future Biodefense 

research agenda, which is articulated in this document. Specifically, the panel was 

asked to provide the following guidance to NIAID: 

•	 Assess the current research sponsored by NIAID related to the 

development of effective measures to counter the health consequences 

of bioterrorism. 

•	 Identify goals for the highest priority areas for immediate, intermediate, and 

long-term research related to Biodefense. 

• Make recommendations on the role of NIAID in achieving these priorities. 

The panel included researchers from academic centers and private industry as 

well as representatives from government, civilian agencies, and the military. 

(See Appendix A.) Participants were selected for their scientific expertise on the 

infectious agents considered to be the major bioterrorist threats, their scientific 

leadership, and their broad research experience. An opening plenary session 

described the state of the science for each microbe identified by the Centers for 

Disease Control and Prevention (CDC) as a Category A Agent of Bioterrorism.* 

Panel members then divided into breakout groups to discuss each organism in 

more detail. 

The panel commended NIAID for developing a Biodefense research agenda and 

was supportive of the planned goals. The content of the research agenda that 

follows reflects the panel’s recommendations. Specific goals for each pathogen 

are articulated within each chapter and include priorities for immediate as well as 

intermediate/long-term research. The introduction includes general 

recommendations that are applicable for all areas of NIAID Biodefense research. 

* The Centers for Disease Control and Prevention (CDC) Category A Agents of Bioterrorism include anthrax, smallpox, plague, 

botulism, tularemia, and viral hemorrhagic fevers. (http://www.bt.cdc.gov/Agent/Agentlist.asp#categoryadiseases) 

v 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

INTRODUCTION


As concern grows about the use of biological agents in acts of terrorism or war, 

Federal health agencies are evaluating and accelerating measures to protect the 

public from the health consequences of such an attack. In 1996, when NIAID 

made public its Research Agenda for Emerging Diseases, intentional introduction 

of infectious agents was not discussed explicitly. However, recent events have 

reminded us that bioterrorism can be a major contributor in disease emergence. 

Our ability to detect and prevent infections related to bioterrorist incidents depends 

to a large degree on the state of biomedical science. Basic and applied research 

supported by the National Institutes of Health (NIH) complements the efforts of 

other Federal agencies by developing the essential tools—diagnostics, therapeutics, 

and vaccines—that are needed by physicians, nurses, epidemiologists, and other 

public health workers to prevent and control a disease outbreak. NIAID is the 

primary NIH Institute that supports and conducts research on the diagnosis, 

prevention, and treatment of infections caused by a wide variety of emerging 

pathogens, including agents that could be intentionally introduced. 

The Institute has developed a Strategic Plan for Biodefense Research at the 

National Institute of Allergy and Infectious Diseases (NIAID), which outlines plans 

for addressing research needs in the broad area of bioterrorism and emerging 

and reemerging infectious diseases. (See Appendix B.) This Biodefense 

Research Agenda supplements the strategic plan and articulates the goals for 

research on anthrax, smallpox, plague, botulism, tularemia, and viral hemorrhagic 

fevers. The research agenda focuses on the need for basic research on the 

biology of the microbe, the host response, and basic and applied research aimed 

at the development of diagnostics, therapeutics, and vaccines against these 

agents. In addition, the agenda addresses the research resources, facilities, and 

scientific manpower needed to conduct both basic and applied research on these 

agents. 

1 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

AREAS OF RESEARCH EMPHASIS


The following areas have been identified as priorities for Biodefense research. 

Biology of the Microbe 

Research into the basic biology and disease-causing mechanisms of pathogens 

underpins efforts to develop interventions against agents of bioterrorism. NIAID 

supports research to better understand the microbial components that define a 

pathogen’s life cycle, as well as the events or processes that are critical to initiating 

infection or influencing the severity of disease. This knowledge is crucial to the 

development of preventative and therapeutic strategies. An important tool in 

understanding the basic biology of pathogenic microbes, including potential agents 

of bioterrorism, is the ability to rapidly obtain genome-sequencing information 

about these agents. The application of genomics research, coupled with other 

biochemical and microbiological information, is expected to facilitate the discovery 

of new targets for diagnostics, drugs, and vaccines. Comparative genomics 

(comparing the sequences of different strains of particular organisms) will be a 

particularly important component of future research, helping us to understand 

virulence and pathogenicity factors. 

Host Response 

Research into both innate and adaptive immune responses is critical in the 

development of interventions against agents of bioterrorism. We must define 

pathogen-triggered protective and deleterious immune responses to inform the 

development of products aimed at controlling and preventing disease. The 

identification of innate immune receptors and the functional responses that they 

trigger will enable targeted activation of the innate immune response and induction of 

specific adaptive immunity. Only one adjuvant (alum) is currently approved for 

general vaccine use. Therefore, we need new immunostimulatory agents. To 

develop new tools for disease control, we need a more detailed understanding of 

signaling pathways, of compartmentalization of immune responses, and of pathogenspecific immunoregulation. Little is known about the ways that immune responses 

can vary among different individuals according to age, general health status, 

treatment with immunosuppressive drugs, and genetic makeup. An enhanced 

understanding of these variables and their impact on immunity is critical for the 

design and development of effective vaccines and immunotherapeutics. 

2 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Vaccines 

Vaccines are the most effective method of protecting the public against infectious 

diseases. New and improved vaccines against agents of bioterrorism must be 

suitable for civilian populations of varying ages and health status. In addition, 

vaccines developed to counter civilian bioterrorist attacks must be safe, easy to 

administer, and capable of an immediate protective and/or transmission-blocking 

immune response. Scientists must develop and characterize adjuvants that can 

enhance these desirable characteristics. Basic research on the host immune 

response, including correlates of protection and the basic biology of the microbe, 

will enhance vaccine development efforts. The initial focus at NIAID has been on 

the development of vaccines against smallpox and anthrax. However, the 

development of candidate vaccines for other category A agents remains a priority. 

A critical component of efforts to achieve these goals is enhanced linkages and 

partnerships with industry. 

Therapeutics 

In the event of a bioterrorism incident, we will need effective therapeutics to address 

the immediate health needs of the public. Antimicrobial agents for treating many 

infectious diseases currently exist. However, we need a broader, more robust 

arsenal of anti-infective agents to treat the broad civilian population and to intervene 

against drug-resistant variants that may emerge. Detailed knowledge of the 

pathways that are essential for replication and pathogenesis will enhance drug 

development. However, many diseases caused by category A agents do not provide 

adequate economic incentives for the development of therapeutic or preventive 

measures by industry. Therefore, collaboration between industry and government 

will be needed to produce the next generation of anti-infectives. Another priority is 

the development of monoclonal antibodies with activity against toxins or other critical 

virulence factors. 

Diagnostics 

A successful response to a bioterrorist threat requires diagnostics that can identify 

the pathogen involved. However, the initial clinical signs and symptoms of many 

agents considered biothreats are nonspecific and resemble those of common 

infections. The ability to rapidly identify the introduction of a bioterrorism organism 

3 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

or toxin will require diagnostic tools that are highly sensitive, specific, inexpensive, 

easy to use, and located in primary care settings. Ideally, these tests also could 

be used to evaluate the possible spectrum of antimicrobial resistance. We have 

the theoretical means to design and develop such assays. For example, 

microchip-based platforms containing thousands of microbial signature profiles 

could be developed. A centralized database could be constructed to collect this 

information and allow the rapid identification of unusual patterns or clustering 

anywhere in the country. Functional genomic tools to identify multiple organisms 

simultaneously will be important, as will enhanced linkages and partnerships with 

industry. 

Research Resources 

Basic research and the development of new vaccines, therapeutics, and diagnostics 

depend on the availability of research resources. Among the resources needed to 

conduct Biodefense research are genetic, genomics, and proteomics information; 

appropriate in vitro and animal models; validated assays to measure immune and 

other host responses; and standardized reagents. In addition, research with 

potentially deadly category A agents must be conducted in appropriate containment 

facilities and must follow procedures that eliminate the threat to laboratory and 

clinical personnel and adjacent communities. Access to biosafety level (BSL) 3/4 

facilities, particularly those with the capacity for animal model and clinical research, is 

limited and must be expanded. Expertise in a wide variety of areas, including 

structural biology, ecology, medicinal chemistry, bioinformatics, the development and 

humanization of antibodies, diagnostic validation, and therapeutic and vaccine 

candidate production, is needed. NIAID understands that a “goal oriented approach” 

to developing countermeasures against bioterrorism agents will require new 

collaborations between basic scientists, process development scientists, and 

engineers to quickly transition vaccine and therapeutic candidates into clinical 

testing. In addition, enhanced relationships among NIH, academia, and industry will 

be necessary to produce clinical material that meets good clinical practice 

requirements. NIAID’s research agenda will include the training of a new cohort of 

investigators, the physical infrastructure within which to conduct this research and 

production, and the technologies, animal models, and reagents necessary to pursue 

this line of research and clinical testing. 

4 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

GENERAL RECOMMENDATIONS


The following recommendations apply to all areas of NIAID counterbioterrorism research. 

Immediate Research 

•	 Develop regional Centers of Excellence for Bioterrorism and Emerging 

Diseases Research. 

•	 Expand the capacity to conduct phase I, II, and III evaluations of candidate 

vaccines and treatments for agents of bioterrorism. 

•	 Expand nonhuman primate capability to evaluate new therapeutic and 

vaccine products. 

•	 Attract new scientific disciplines to Biodefense research, and expand the 

research training of a new cohort of investigators. 

•	 Expand extramural and intramural research and clinical infrastructure, 

including construction and renovation of BSL-3/4 laboratories. 

• Expand the availability of animal models for preclinical research. 

•	 Develop rapid, inexpensive, and broad-based clinical diagnostics 

approaches using genomics and proteomics. 

•	 Encourage structural genomics and proteomics research for the targeted 

development of drugs, vaccines, and diagnostics. 

•	 Encourage industry participation to ensure the availability of rapid, 

sensitive, and licensed diagnostics to hospital clinical laboratories. 

• Expand partnership opportunities with other agencies and governments. 

•	 Develop a centralized repository for reagents and clinical specimens for 

agents of bioterrorism. 

•	 Develop procedures and cGMP facilities capable of producing 

monoclonal antibodies, vaccines, and other immunotherapies for phase 

I and II clinical studies. 

5 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

• Enhance adjuvant discovery and rational design of Toll system mediators. 

•	 Identify and characterize innate and adaptive immune responses that occur 

after exposure to agents of bioterrorism and enhance basic research on 

mucosal immunology. 

•	 Establish MHC/peptide and B-cell epitope databases that may be used to 

further define immune responses, including the identification of relevant 

immune polymorphisms, and maximize such responses. 

Intermediate and Long-Term Research 

•	 Examine human genetic variation in response to therapy, in drug 

susceptibility, and in infectivity. 

• Define the human microbiome in states of health and disease. 

•	 Develop predictive immunology tools that will allow for the design of 

interventions for currently unknown biological weapons. 

•	 Analyze unique responses at the genome level to infection with agents of 

bioterrorism. 

• Analyze gene expression of agents of bioterrorism in vivo. 

• Standardize and validate protocols that involve studies using animal models. 

• Expand clinical capacity for phase I safety studies. 

•	 Develop, test, and analyze novel vaccination and protective strategies in 

all populations (e.g., the very young, the elderly, and 

immunocompromised individuals). 

•	 Enhance currently available international capability by expanding existing 

clinical trial networks (e.g., the International Collaborations in Infectious 

Disease Research Program). 

•	 Address human resource needs required for supporting research through 

training grants, fellowships, travel grants, and faculty development. 

6 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

•	 Define the genetic basis of host susceptibility to infection with agents 

of bioterrorism. 

•	 Identify pathogen-induced immunoregulatory and immunosuppressive 

effects that influence the host’s ability to mount an effective immune 

response and respond to postexposure vaccination or immunotherapy. 

7 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

ANTHRAX


Bacillus anthracis, the agent that causes anthrax, has several characteristics that 

make it a formidable bioterrorist threat. These characteristics include its stability in 

spore form, its ease of culture and production, its ability to be aerosolized, the 

seriousness of the disease it causes, and the lack of sufficient vaccine for 

widespread use. 

Human anthrax has three major clinical forms: cutaneous, inhalational, and 

gastrointestinal. If left untreated, all three forms can result in septicemia and 

death. Early antibiotic treatment of cutaneous and gastrointestinal anthrax is 

usually curative; however, even with antibiotic therapy, inhalational anthrax is a 

potentially fatal disease. Although case-fatality estimates for inhalational anthrax 

are based on incomplete information, the historical rate is considered to be high 

(about 75 percent) for naturally occurring or accidental infections, even with 

appropriate antibiotics and all other available supportive care. However, the 

survival rate after the recent intentional exposure to anthrax in the United States 

was 60 percent for the first 10 cases. 

Biology of the Microbe 

Inhalational anthrax develops after spores are deposited in alveolar spaces and 

subsequently ingested by pulmonary alveolar macrophages. Surviving spores are 

then transported to the mediastinal lymph nodes, where they may germinate up to 

60 days or longer. After germination, replicating bacteria release toxins that result 

in disease. Major virulence factors include an antiphagocytic outer capsule and at 

least two well-characterized toxins. The two toxins, called edema factor (EF) and 

lethal factor (LF), can destroy cells or inhibit their normal functioning. A third 

component, called protective antigen (PA), when associated with both EF and LF, 

enables EF and LF to bind to a specific receptor on mammalian cells. After this 

complex is internalized, the bacteria’s toxic effects are activated. Researchers 

recently engineered mutant recombinant PAs (rPAs) that bind to the native 

receptor. These mutant rPAs also can displace wild-type PA by blocking and 

interrupting the delivery of LF and EF into cells. Recent studies also have 

identified the region of the mammalian cell receptor to which PA binds and have 

determined the structure of the LF binding site. Soluble fragments of the receptor 

containing the toxin-binding site can function as decoys to protect cells from 

damage by LF. Other recent studies have characterized the site where LF binds to 

8 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

MAPKK (mitogen-activated protein kinase kinase), a vital intracellular enzyme 

whose disruption by LF causes cell death. 

Sequencing of the chromosomal genome of B. anthracis is nearly completed. 

The genes for LF, EF, and PA are contained on plasmids that already have been 

sequenced. NIAID is expanding sequencing efforts with a comprehensive 

genomic analysis of B. anthracis and related bacilli. Researchers will use 

sequence data derived from selected strains, isolates, and related species to 

assess the degree of genetic variation and diversity. This genetic information will 

provide a framework in which to evaluate the basis for differences in pathogenicity 

and virulence that have been noted between strains. Other uses for the genomic 

data include supporting basic research to identify specific molecular markers and 

targets for strain identification and molecular genotyping; developing sequencebased detection technologies; and designing effective vaccines, therapies, and 

diagnostic tools. In addition, the data will enhance the detection of genetic 

polymorphisms that correlate with phenotypes, such as drug resistance, morbidity, 

and infectivity, as well as key events or processes that influence the germination 

of spores in vivo. A comprehensive bioinformatics resource will support and 

maintain microbial genomic databases and the development of associated 

software and bioinformatics tools. These approaches will serve as a prototype for 

other microorganisms with potential to be used as agents of bioterrorism. 

Host Response 

Individuals who contract anthrax produce IgG1 and IgG3 anti-PA specific antibodies. 

Vaccination with the UK human vaccine generates primarily IgG1 antibodies, but 

IgG3, IgG2, and IgG4 antibodies also are detectable. Protection in animal models 

correlates with anti-PA antibody production. Recent evidence suggests that spore 

components also induce antibody production in rabbits, monkeys, and guinea pigs 

that have been vaccinated with an unencapsulated, toxigenic live vaccine strain. 

Sera from these vaccinated animals exhibit two antispore activities associated with 

antibody function (i.e., inhibition of spore germination and stimulation of spore 

phagocytosis by macrophages). Antibiotic therapy started after initial infection does 

not interfere with the immune response to B. anthracis. These findings indicate that it 

may be feasible to design a postexposure vaccination strategy to protect against the 

persistence of small numbers of spores in the lungs after antibiotic therapy. 

9 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Vaccines 

Anthrax vaccine adsorbed (AVA), prepared from a cell-free filtrate of a 

nonencapsulated attenuated strain of B. anthracis, is the only licensed anthrax 

vaccine. PA of the anthrax toxin is the main component responsible for generating 

immunity. The Department of Defense (DoD) is using AVA in its mandatory 

immunization program to protect U.S. military personnel in high-threat areas. 

CDC recommends civilian use of AVA only for high-risk populations, such as 

veterinarians and laboratory workers. The schedule for administering this 

vaccine—six doses over the course of 18 months—is too protracted for use in 

response to a bioterrorism incident. Therefore, efforts are underway to devise an 

abbreviated, yet effective, postexposure immunization schedule. 

Ensuring the safety of very young, elderly, and immunocompromised 

individuals requires a different vaccine regimen than that used for the military. 

NIAID has been collaborating with DoD to support the development of nextgeneration anthrax vaccines that may be more appropriate than AVA for use in 

civilians and military personnel. A vaccine based on a rPA appears to be the 

most promising candidate. 

Although the correlates of immunity have not yet been defined, several animal 

studies have shown that neutralizing antibodies to PA elicit significant protective 

immunity against inhalational spore challenge. In vitro studies have shown that 

antibodies to PA also recognize spore coat proteins, stimulate the uptake of spores 

by phagocytes, and inhibit spore germination. 

Therapeutics 

NIAID is exploring the use of alternative antimicrobials or antitoxin therapies for 

anthrax. Together with the Food and Drug Administration (FDA), CDC, and DoD, 

NIAID is working to prioritize and accelerate testing of additional antimicrobial 

candidates for inhalational anthrax. NIAID-supported investigators recently 

published the results of two studies on the mechanisms by which anthrax toxins 

destroy cells. In one study, scientists identified the site at which PA binds on 

mammalian cells as well as a compound that can interfere with binding of the 

toxin. In the other study, researchers determined the structure of the LF 

component of the anthrax toxin. The information gained through these studies will 

likely hasten the development of new drugs to treat anthrax. 

10 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

For the treatment of inhalational anthrax in humans, FDA has approved both


penicillin G procaine and doxycycline. Ciprofloxacin also has shown efficacy in


animal studies and recently received a label indication for inhalational anthrax.


Current CDC recommendations for treating inhalational anthrax include initial


intravenous therapy with ciprofloxacin or doxycycline and one or two additional


antibiotics; patients may be switched to oral antibiotics when clinically appropriate.


Treatment should continue for 60 days. Cephalosporins are not an effective


treatment because B. anthracis isolates produce a cephalosporinase. 


However, recent cases of inhalational anthrax indicate that the combination of a


fluoroquinolone with at least one other active drug may improve efficacy. 


Diagnostics 

Rapid diagnostic tests, such as enzyme-linked immunosorbent assay (ELISA) and 

polymerase chain reaction (PCR) for PA, are available at national and state 

reference laboratories. However, these tests are not licensed. They are used 

primarily as confirmation of a clinical diagnosis because of the time required to 

transport samples and complete testing. Culture remains the standard 

confirmatory test, but definitive identification of B. anthracis requires additional 

testing beyond a standard blood culture. In the recent intentional exposures to 

anthrax, organisms could not be cultured from samples obtained after the initiation 

of antibiotic treatment. Several transportable, rapid assays are being developed 

for use in humans and veterinary practice. 

Research Resources 

Several pressing resource needs must be addressed to ensure a robust research 

program on anthrax. Although most animal species are susceptible to anthrax, 

mice are generally used in experimental studies because of their availability and 

cost. Nonhuman primates, rabbits, and guinea pigs are the animal models used in 

inhalational spore challenge studies. All animal evaluations that involve infectious 

strains of anthrax will require adequate and secure BSL-3 facilities. In addition, 

any clinical studies that involve infected patients will require appropriate facilities. 

Attracting new and experienced investigators into anthrax research also will require 

appropriate support in terms of facilities, reagents, and training. 

11 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Goals 

The following goals are priorities for Biodefense research on anthrax. 

Immediate Research Goals 

•	 Establish capacity for the development and production of pilot lots of 

candidate anthrax vaccines. 

•	 Conduct phase I and II trials with rPA anthrax vaccine candidates and 

alternative adjuvant formulations. 

•	 Expand clinical capability to accelerate phase I and II testing of candidate 

anthrax vaccines. 

•	 Establish a centralized immunology laboratory to develop and validate tests 

required for the licensure of anthrax vaccines. 

•	 Screen existing FDA-approved antimicrobials and immunomodulators for 

efficacy against anthrax. 

•	 Encourage exploration of new targets for antimicrobial therapies, including 

strategies to prevent germination of spores, to synthesize or neutralize 

toxins, and to interfere with the attachment and entry of toxins into host 

target cells. 

•	 Conduct comparative genomic sequencing of selected Bacillus strains to 

detect subtle differences in pathogenesis and virulence associated with 

antigens or other factors. 

• Develop and evaluate in vivo transmission and spore germination models. 

• Identify and characterize new virulence and pathogenicity factors. 

•	 Identify targets within innate and adaptive pathways that can be used to 

modulate infection. 

•	 Identify and characterize innate and adaptive immune responses that 

occur after initial exposure to anthrax, including responses associated with 

spore germination. 

12 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Intermediate and Long-Term Research Goals 

•	 Determine the surrogates of immunity for rPA vaccines using nonhuman 

primate challenge models. 

•	 Identify new therapeutics by translating basic research findings into targets 

for intervention, for the development and optimization of monoclonal and 

polyclonal antibodies, and for new immunotherapeutics. 

•	 Synthesize active lead compounds in sufficient quantities for preclinical 

pharmacokinetics and studies of animal model efficacy, mechanism of 

action, and toxicology. 

•	 Use genomic-sequencing information to conduct comparative genomic 

analysis and to identify genetic variations and potential targets for vaccines 

and treatments against anthrax. 

•	 Develop robust, sensitive, and specific diagnostics for the detection of 

anthrax antigens in clinical specimens. 

•	 Explore the use of microbial signatures in gene expression profiles of 

host cells. 

• Develop technologies for in situ studies of anthrax. 

•	 Continue to improve anthrax vaccines by focusing basic research on the 

identification of potential targets, the development and optimization of 

vaccine delivery systems, the optimization of B- and T-cell protective 

responses, the induction of long-term immunologic memory, and the 

development of adjuvants based on innate immunity. 

13 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

SMALLPOX


Smallpox, which is caused by the virus Variola major, is considered one of the 

most dangerous potential biological weapons because it is easily transmitted from 

person to person, no effective therapy exists, and few people carry full immunity to 

the virus. Although a worldwide immunization program eradicated smallpox 

disease in 1977, small quantities of smallpox virus still exist in two secure facilities 

in the United States and Russia. However, it is likely that unrecognized stores of 

smallpox virus exist elsewhere in the world. 

The symptoms of smallpox infection appear approximately 12 days (the range is 

from 7 to 17 days) after exposure. Initial symptoms include high fever, fatigue, 

headache, and backache. A characteristic rash, which is most prominent on the 

face, arms, and legs, follows in 2 to 3 days. The rash starts with flat red lesions 

(a maculopapular rash) that evolve into vesicles. Unlike chickenpox, the lesions 

associated with smallpox evolve at the same rate. Smallpox lesions become filled 

with pus and begin to crust early in the second week after exposure. Scabs 

develop, separate, and fall off after approximately 3 weeks. Individuals are 

generally infectious to others from the time immediately before the eruption of the 

maculopapular rash until the time scabs are shed. Smallpox spreads directly from 

person to person, primarily by aerosolized saliva droplets expelled from an infected 

person. Contaminated clothing or bed linens also can spread the virus. The 

mortality of smallpox infection is approximately 30 percent, and patients who 

recover frequently have disfiguring scars. 

Biology of the Microbe 

The variola virus has not been well studied because of the hazards associated 

with potential exposure. However, vaccinia virus, which is used as a smallpox 

vaccine and is closely related to variola, is well studied. The few comparative 

studies of the two viruses have shown that the major differences are in the host 

ranges: whereas vaccinia infects several hosts, variola infects only humans 

naturally and cynomolgus monkeys under artificial laboratory conditions. The two 

viruses can be distinguished by the appearance of lesions on chick embryo 

chorioallantoic membranes and by tissue culture growth characteristics. The 

viruses share antigens and generate cross-neutralizing antibodies, a characteristic 

that has been exploited in the use of the vaccinia vaccine to prevent smallpox. 

The two viruses can be distinguished by PCR, ELISA, radioimmunoassays, and 

14 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

monoclonal antibodies. Vaccinia is now being investigated extensively as a vector 

for the delivery of other vaccine genes. 

Two forms of infectious orthopoxvirus are produced in infected cells: intracellular 

mature virus (IMV) that remain in the infected cell and extracellular enveloped virus 

(EEV) that are released from the cell late in infection. The EEV form of the virus 

contains an additional lipid envelope and cellular and viral proteins, thus making 

EEV immunologically different from IMV. In addition, the EEV and IMV forms enter 

cells by different mechanisms, use different cell receptors, and have different 

sensitivities to antibodies and complement. Immune evasion by poxviruses is 

accomplished through mechanisms related to the release of proteins that bind 

chemokines, EEV resistance to neutralizing antibodies, and EEV resistance to 

complement destruction through acquisition of host complement control proteins. 

Variola and vaccinia belong to the Orthopoxvirus genus of poxviruses. These 

double-stranded DNA viruses replicate in the cytoplasm, unlike other DNA viruses 

that depend on host nuclear DNA replication enzymes. Several strains of variola 

and vaccinia have been genomically sequenced. The genes for structural, 

membrane, and core proteins appear to be highly conserved among 

orthopoxviruses. Genes responsible for growth in human cells also have been 

identified. NIAID will actively pursue further research in these areas. 

Host Response 

Sparse data are available on the duration of protection that vaccinia immunization 

confers against natural smallpox infection. However, it is known that natural 

infection confers life-long protection. After a smallpox outbreak in Europe, the 

case-fatality rate was 52 percent in unvaccinated people and 1.4 percent in 

people who were vaccinated up to 10 years previously. However, the casefatality rate rose to 11.4 percent in people who were vaccinated more than 20 

years previously. Individuals who recover from natural infection with variola or 

are immunized with vaccinia develop neutralizing antibodies. These antibodies 

are long-lived and can be detected up to 20 years after vaccination. Because the 

science of cellular-mediated immunity was in its infancy at the time that smallpox 

vaccination was discontinued, few data are available about the type and duration 

of both CD4 and CD8 T-cell responses or their correlation with immunity. Viral 

core and envelope proteins induce antibody responses in humans, but no specific 

15 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

B-cell, CD4, or CD8 T-cell epitopes have been mapped. Delayed type 

hypersensitivity, a measure of T cell-mediated immunity, begins to develop as 

early as 2 days after vaccinia vaccination. 

Vaccines 

Smallpox vaccine consists of a laboratory strain of vaccinia virus and has 

proven to be highly effective in preventing infection. It is believed that in 

unvaccinated people exposed to smallpox, the vaccine can lessen the severity 

of, or even prevent, illness if given within 4 days of exposure. 

Vaccination to prevent smallpox has not been required in the United States 

since 1972. People vaccinated before 1972 very likely have diminished 

immunity to smallpox; people born in the United States after 1972 have not 

been routinely vaccinated. Currently, smallpox vaccination is recommended 

and available only for individuals who are at risk of imminent exposure, such as 

laboratory personnel who work with vaccinia and other orthopoxviruses related 

to the smallpox virus. The smallpox vaccine is highly reactogenic and often 

causes fever as well as large local reactions. In addition, the vaccine is 

associated with several complications, some of which are life threatening, 

particularly in people with immune deficiencies. 

A bioterrorist attack involving smallpox, or the accidental release of the virus 

from a clandestine laboratory, would require using stores of the existing smallpox 

vaccine. Approximately 15 million doses of the FDA-approved Dryvax vaccine 

have been stored since production stopped in 1983. This supply probably would 

not be enough to respond to a national smallpox epidemic. As an immediate 

response to this vulnerability, NIAID initiated a study to determine the feasibility 

of diluting Dryvax vaccine to make more doses available from the existing 

stores. In this study, investigators examined the skin and immune system 

responses of normal, unimmunized adult volunteers who were given a 1:10 

dilution or a 1:100 dilution of Dryvax vaccine. Researchers compared the 

responses to those of other volunteers who had received the full-strength 

vaccine. The results demonstrate that the full-strength vaccine has maintained its 

potency. In addition, 70 percent of the people who received a single dose of the 

10-percent vaccine developed a sore followed by a scab at the injection site and 

antibodies in their blood, both signs that the vaccine had conferred protection. 

16 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

A larger study to compare 1:5 and 1:10 dilutions with the full-strength Dryvax 

vaccine is now underway. 

Therapeutics 

In collaboration with DoD, NIH has screened more than 500 compounds 

against smallpox and related viruses. Compounds were selected for antiviral 

testing from the following categories: FDA-approved drugs that are effective 

against other viruses, antiviral compounds that are in clinical development for 

other viruses, known experimental antiviral compounds, and new chemical 

entities. In addition, scientists are evaluating some drugs approved by FDA for 

nonviral indications. In vitro evaluation has shown that several drugs tested in 

these screening studies, including a number of cidofovir derivatives, are active 

against both vaccinia and cowpox. 

Diagnostics 

Most smallpox cases present with a characteristic rash that is denser on the 

extremities and face than on the torso. Confirmatory tests include electron 

microscopy of vesicular or pustular fluid or virus culture, PCR analysis of 

peripheral blood mononuclear cells (PBMCs) or vesicle fluid, or restriction 

fragment length polymorphisms (RFLPs). 

Research Resources 

NIAID supports evaluation of experimental antiviral compounds in a number of 

localized and systemic mouse models of vaccinia and cowpox. DoD also 

supports antiviral evaluations in similar rodent systems. Both resources 

increase the total capacity for mouse model evaluation and allow for 

independent confirmation of positive results. NIAID also supports a 

mouse/ectromelia model and a vaccinia model in rabbits. Compounds that are 

effective in these small-animal models are prioritized for evaluation in the 

monkeypox primate model by DoD investigators. Several pressing resource 

needs must still be addressed to ensure a robust research program on 

vaccinia/cowpox models. Attracting new and experienced investigators into 

research on surrogate models of smallpox also will require appropriate support 

in terms of facilities, reagents, and training. 

17 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Goals 

The following goals are priorities for Biodefense research on smallpox. 

Immediate Research Goals 

•	 Expand the existing supply of live and attenuated vaccines, with particular 

emphasis on vaccines with reduced reactogenicity. 

• Initiate and expand clinical trials of existing smallpox vaccines. 

•	 Determine the correlates of immunity for smallpox vaccines through the 

detailed evaluation of immune responses to Dryvax vaccine. 

•	 Conduct phase I and ll trials with new candidate smallpox vaccines, with 

particular emphasis on the cell culture vaccines currently under development. 

•	 Develop a centralized immunology laboratory to validate assays required 

for the licensure of smallpox vaccines. 

•	 Fully characterize cidofovir’s activity against poxviruses and ensure that an 

adequate supply of the drug is available to treat complications from 

vaccination. 

• Develop animal models for studying smallpox pathogenesis. 

•	 Expand in vitro and in vivo screening capability for oral antivirals, 

immunotherapies, and replacements for vaccinia immune globulin. 

•	 Validate current diagnostics for orthopoxvirus infections and other vesicular 

rashes (i.e., chickenpox). 

•	 Conduct comparative genomic sequencing of additional pox virus strains to 

detect potential differences in pathogenesis and virulence. 

•	 Identify and characterize host factors and viral proteins that are involved in 

the production and maintenance of the two forms of infectious 

orthopoxviruses, IMV and EEV. 

18 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Intermediate and Long-Term Research Goals 

•	 Develop detailed innate and adaptive immunologic profiles of responses to 

orthopox vaccines. 

•	 Understand and prevent complications of smallpox vaccination, such as 

eczema vaccinatum and vaccinia gangrenosa. 

•	 Develop animal model capability and provide the required standardization 

and validation for the development of vaccines and therapeutics against 

smallpox. 

•	 Develop and optimize human monoclonal and polyclonal antibodies as 

potential therapies against smallpox. 

•	 Expand basic research on host-virus interactions, viral replication, and 

immune evasion mechanisms. 

• Investigate the effect of infection on cytokine production. 

19 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

PLAGUE


Plague is caused by the bacterium Yersinia pestis. Its potential for use as a


biological weapon is based on methods that were developed to produce and


aerosolize large amounts of bacteria and on its transmissibility from person to


person in certain of its forms. An additional factor is the wide distribution of


samples of the bacteria to research laboratories throughout the world. Infection by


inhalation of even small numbers of virulent aerosolized Y. pestis bacilli can lead to


pneumonic plague, a highly lethal form of plague that can be spread from person


to person. Natural epidemics of plague have been primarily bubonic plague, which


is transmitted by fleas from infected rodents. 


Symptoms of pneumonic plague, including fever and cough, resemble those of


other respiratory illnesses such as pneumonia. Symptoms appear within 1 to 6


days after exposure and lead rapidly to death. If untreated, pneumonic plague has


a mortality rate that approaches 100 percent. Antibiotics are effective against


plague, but an effective vaccine is not widely available.


Biology of the Microbe 


Although Y. pestis is very efficient at invading host epithelial cells, the molecular


mechanisms that contribute to its invasiveness are not understood. Various iron


transport mechanisms as well as the interaction of at least three quorum-sensing


mechanisms appear to be involved. 


Because the genome of Y. pestis has been completely sequenced, it should be


possible to accelerate efforts to characterize key events in pathogenesis that will


help identify suitable vaccine candidates, diagnostic reagents, and key targets for


drug intervention. The Y. pestis outer surface membrane proteins (Yomps), of


which there are several, appear to be important virulence factors and play a major


role in pathogenesis. Y. pestis has a set of virulence-associated proteins that are


plasmid encoded. Ambient temperature and Ca++ levels regulate the expression


and secretion of these proteins through the so-called low-Ca++ response (LCR)


mechanism. Further characterization of plasmid-encoded proteins and their role in


pathogenesis could provide the basis for an effective subunit vaccine. 


To cause infection, Y. pestis and other pathogenic bacteria need to remove iron—


an essential trace nutrient—from host iron- and/or heme-chelating proteins. 


Y. pestis has three partially characterized iron transport systems that play an


20 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

important role in iron transport and removal. One of these systems is siderophoredependent and involves the synthesis of yersiniabactin (Ybt). Since the Ybt 

system is essential for iron acquisition during the early stages of plague, it may be 

an excellent target for early intervention and treatment. 

Host Response 

Two Y. pestis proteins, F1 and V, are being evaluated as vaccine candidates in 

animal models. The F1 protein is considered the principle protective antigen in the 

Plague Vaccine USP. Recent protection studies in mice show that injection with 

recombinant F1, V, or F1+V protein adsorbed to an aluminum hydroxide adjuvant 

provided protection against bubonic and pneumonic forms of the disease; similar 

information is not available for humans. The F1+V vaccine provides significant 

protection as early as 14 days after immunization. Antibody titers to F1 and V 

peak at 5 to 12 weeks after immunization and last for at least 1 year. Passive 

transfer of either V protein-specific polyclonal antiserum or a monoclonal anti-V 

antibody protects animals against lethal challenge with Y. pestis. In addition, 

Yersinia outer proteins may be antigenic. Antibodies to some of these proteins 

have been identified in sera from convalescent plague patients and in sera from 

experimentally infected mice. 

Vaccines 

A killed vaccine has been licensed for use in the United States for individuals 

considered to be at high risk for developing bubonic plague, including laboratory 

workers and military personnel in areas where plague is endemic. Vaccination 

before exposure was effective in preventing or reducing disease. However, the 

vaccine did not prevent or reduce disease from pneumonic plague. The vaccine 

has not been produced since 1999 and is no longer available. Clearly, we need to 

develop a new vaccine for use in a preexposure setting or in a postexposure 

setting in conjunction with antibiotic therapy to protect against pneumonic plague. 

The V antigen has been identified as a possible vaccine candidate. However, 

other virulence-associated antigens must be defined to devise effective multivalent 

vaccines to combat the threat of new, genetically engineered resistant strains. 

Therapeutics 

Treatment of plague with antibiotics generally is effective, but success strongly 

depends on how quickly drug therapy is started, the dose of inhaled bacteria, and 

21 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

the level of supportive care for the patient. If treatment is delayed more than 

24 hours after symptoms begin, mortality remains high. A number of drugs, 

including streptomycin, gentamicin, tetracycline, and doxycycline, are recommended 

for use in the treatment of pneumonic plague on the basis of their efficacy in animal 

models of infection. However, the potential use of engineered, multi-drug-resistant 

strains as agents of bioterrorism emphasizes the need to develop alternative 

treatment strategies and antimicrobials with novel mechanisms of action. 

Diagnostics 

A rapid diagnostic test for plague is not widely available, but confirmatory tests are 

available at some state laboratories, CDC, and military facilities. Antibody 

detection assays are not useful for diagnosis because antibodies take several days 

or weeks to develop. The relatively rare occurrence of pneumonic plague in the 

United States has resulted in a low index of suspicion of naturally occurring cases 

and a lack of early-warning sentinel systems to detect aerosol exposure. 

Research Resources 

Several resource needs must be addressed to ensure a robust research program 

on Y. pestis. Because most animal species are susceptible to plague, several 

species have been used in experimental studies. Currently, the mouse is the most 

commonly used animal model of disease. Adequate and secure BSL-2/3 facilities 

will be required for all animal evaluations that involve infectious strains of Y. pestis. 

In addition, any clinical studies that involve infected patients will require 

appropriate facilities. Attracting new and experienced investigators into Y. pestis 

research also will require appropriate support in terms of facilities, reagents, and 

training. 

Goals 

The following goals are priorities for Biodefense research on plague. 

Immediate Research Goals 

• Accelerate the search for candidate Y. pestis vaccines. 

•	 Establish capacity for the development, refinement, production, and testing 

of pilot lots of candidate Y. pestis vaccines. 

•	 Encourage exploration of new targets leading to the development of 

Y. pestis-specific chemotherapeutics and/or entities with novel modes of 

22 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

action (e.g., Ybt, TTSS, LcrV). 

•	 Screen existing FDA-approved antimicrobials and immunomodulators for 

efficacy against Y. pestis. 

•	 Develop rapid, inexpensive, and broad-based clinical diagnostics 

approaches for plague. 

•	 Develop standards for validation and comparison of potential 

plague diagnostics. 

Intermediate and Long-Term Research Goals 

•	 Characterize natural plague foci worldwide by examining factors such as 

microbial ecology and defining infectious reservoirs and vectors. 

• Develop microarrays for functional genomics studies of plague. 

•	 Identify and characterize innate and adaptive immune responses that occur 

after initial exposure to plague. 

•	 Identify inappropriate and immunopathogenic responses to infection with 

plague and strategies for the minimization of such responses. 

• Determine correlates of immunity for Y. pestis. 

•	 Examine host-pathogen interactions and host response to infection 

with plague. 

•	 Identify novel diagnostic and prognostic signatures based on molecular and 

spectral patterns after infections with plague. 

23 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

BOTULISM


Botulinum toxin, which is produced by the spore-forming anaerobic bacterium 

Clostridium botulinum, is a highly toxic substance that presents a major threat from 

intentional exposure. The toxin is highly lethal and easily produced and released 

into the environment. Botulinum toxin is absorbed across mucosal surfaces and 

irreversibly binds to peripheral cholinergic nerve synapses. Seven antigenic types 

(A–G) of the toxin exist. All seven toxins cause similar clinical presentation and 

disease; botulinum toxins A, B, and E are responsible for the vast majority of 

foodborne illnesses in the United States. 

Exposure to the toxin induces symptoms that include muscle paralysis; difficulty in 

speaking, swallowing, or seeing; and, in severe cases, the need for mechanical 

respiration. People exposed to the toxin require immediate and intensive 

supportive care and treatment. The onset and severity of symptoms depend on 

the rate and amount of toxin absorbed into circulation. With foodborne exposure, 

incubation varies from 2 hours to 8 days but is generally limited to 72 hours. 

Symptoms subside when new motor axon twigs reenervate paralyzed muscles, a 

process that can take weeks or months in adults. 

Biology of the Microbe 

C. botulinum does not normally infect humans. However, humans are exposed to 

the toxin after eating food contaminated with the organism. Botulinum toxin’s 

mechanism of action is well understood. The toxin consists of a heavy chain and 

a light chain joined by a single disulfide bond that is essential for neurotoxicity. 

Both the sequence and three-dimensional structure of the toxin have been 

determined. The structure consists of three functional domains: a catalytic subunit, 

a translocation domain, and a binding domain. The toxin binds irreversibly to an 

unidentified receptor on presynaptic membranes of peripheral cholinergic 

synapses, mainly at neuromuscular junctions. After internalization of the toxin and 

translocation into the cytosol, a Zn++-containing endopeptidase on the light chain 

blocks acetylcholine release from motor neurons. The release is blocked by 

preventing fusion of acetylcholine-containing vesicles with the terminal membrane. 

The seven botulinum toxins exhibit somewhat different protease activities, cleaving 

three SNARE proteins (synaptobrevin/ VAMP, SNAP-25, and syntaxin) at different 

sites. The molecular basis of this proteolytic specificity is not fully understood. 

The SNARE proteins are essential in the trafficking of synaptic vesicles to the 

presynaptic membrane. 

24 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Host Response


The botulinum neurotoxins that are produced by C. botulinum consist of seven


immunologic serotypes, A–G. Therapy for human botulism includes passive


immunization with equine antitoxin. The licensed antitoxin, which consists of


neutralizing antibodies to the A, B, and E toxins, is available from CDC. Recent


studies in mice identified antigenic epitopes within toxin A that activate B- and 


T-cell responses. Comparison of the amino acid sequences of toxins A, B, and E


showed considerable homology among the three proteins in some of the defined


antigenic regions. This finding suggests that it may be possible to induce cross-


protective immunity.


Vaccines 

No approved vaccine to prevent botulism exists. However, an investigational 

botulinum toxoid is available in limited quantities to induce immunity in laboratory 

workers. Since immunity develops over several months, the toxoid would not be 

useful for postexposure prophylaxis. Development of a vaccine strategy should 

take into account the therapeutic use of BotoxTM for a number of human conditions 

(e.g., strabismus, spasticity, and cervical dystonia). Vaccination of the larger 

community would negate the effectiveness of BotoxTM treatment. Therefore, it is 

likely that a vaccine would be used only in high-risk groups. 

Therapeutics 

Treatment of botulism consists of supportive care and the use of antitoxins. Timely 

administration of the available antitoxin minimizes subsequent nerve damage and 

disease severity, but it does not appear to alter existing paralysis. An investigational 

equine heptavalent antitoxin (A–G reactive), which covers all seven toxins, is being 

developed by DoD. The safety of this antitoxin has not been established. Although 

available data are limited, they do not indicate that the antitoxin poses safety 

concerns for special populations, such as pregnant women, older adults, and young 

children. Antibiotics are not effective against botulinum toxin, although clindamycin 

has been reported to have activity against one of the toxins. 

25 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Diagnostics 

Botulism is frequently misdiagnosed. Clinical diagnosis must be confirmed with a 

mouse bioassay, which is based on neutralization by specific antitoxins. This 

assay takes up to 2 days to complete and is available in only a few laboratories. A 

rapid and sensitive diagnostic for botulism is essential for treatment because 

samples must be obtained before antitoxin treatment is begun. 

Research Resources 

Unlike several other agents of bioterrorism, botulinum toxin is not an infectious 

agent. Although highly lethal, it does not require specialized facilities to ensure 

containment. However, BSL-3 procedures conducted in BSL-2 facilities are 

recommended for work with botulinum toxin. Because most animal species are 

susceptible to botulism, several species have been used in experimental studies. 

Currently, the mouse is the most commonly used animal model of disease. 

Attracting new and experienced investigators to botulinum toxin research will 

require appropriate support in terms of reagents and training. 

Goals 

The following goals are priorities for Biodefense research on botulism. 

Immediate Research Goals 

•	 Process, produce, and conduct phase I and II trials with the heptavalent 

equine antitoxin. 

•	 Scale up production and phase I testing of three human monoclonal 

antibodies to toxin A. 

• Develop and test human monoclonal antibodies to toxins B, E, C, F, G, and D. 

•	 Produce and conduct phase I and II trials of recombinant toxin B vaccine 

fragment C. 

• Develop recombinant fragment C vaccine against toxins A, F, E, C, G, and D. 

•	 Develop rapid and inexpensive diagnostics for botulism toxins and their 

genes for use in multiple settings. 

•	 Develop effective cell culture systems to study toxin binding, internalization, 

and protease activity. 

26 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Intermediate and Long-Term Research Goals 

•	 Identify toxin receptors on both mucosal transport cells (digestive and 

respiratory tracts) and on human cholinergic cells. 

• Clone, overexpress, and determine the structure of toxin receptors. 

•	 Develop specific pharmacologic agents as toxin-binding inhibitors for 

mucosal and nerve cells. 

• Characterize the molecular basis of proteolytic specificity of botulinum toxins. 

•	 Conduct basic research on the mechanisms of action of the seven toxin 

serotypes, including receptor-binding specificity and proteolytic 

substrate specificity. 

• Develop small molecule inhibitors of toxin protease activity. 

•	 Develop systems for the delivery of small molecule inhibitors to cholinergic 

nerve cells. 

27 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

TULAREMIA


Tularemia is a potential bioterrorist agent because of its high level of infectivity (as


few as 10 organisms may cause disease) and its ability to be aerosolized.


Francisella tularensis, which causes tularemia, is a non-spore-forming, facultative


intracellular bacterium that can survive at low temperatures for weeks. Two strains


of the organism have been characterized—type A, which is found in North


America, is more virulent than type B, which is found in Europe and Asia. The


disease is not transmitted from person to person; it spreads naturally from small


mammals or contaminated food, soil, or water to humans. Natural infection occurs


after inhalation of airborne particles. 


Tularemia can take one of several forms, depending on the route of exposure.


The disease resulting from the inhalation of airborne F. tularensis is the most likely


intentional exposure. The inhalation form is an acute, nonspecific illness beginning


3 to 5 days after respiratory exposure; in some cases, pleuropneumonia develops


after several days or weeks. If untreated, the disease could lead to respiratory


failure. Treatment with antibiotics reduces mortality for naturally acquired cases by


2 to 60 percent. A live attenuated tularemia vaccine developed by DoD has been


administered under an IND (investigational new drug) application to thousands of


volunteers. To date, use of this vaccine has been limited to laboratory and other


high-risk personnel.


Biology of the Microbe 

The fundamental mechanisms involved in virulence and pathogenesis are not 

known. The cell wall of F. tularensis is unusually high in fatty acids. Loss of the 

capsule may lead to loss of virulence but not viability; however, the capsule is 

neither toxic nor immunogenic. Infection with F. tularensis involves the 

reticuloendothelial system and results in bacterial replication in the lungs, liver, and 

spleen. After respiratory exposure, F. tularensis infects phagocytic cells, including 

pulmonary macrophages. In the liver, F. tularensis has been shown to invade and 

replicate in hepatocytes. Destruction of infected hepatocytes results in the release 

of bacteria and subsequent uptake by phagocytes. When lysis of hepatocytes was 

prevented by the administration of a monoclonal antibody, bacteria continued to 

replicate in the hepatocytes, leading to rapid lethality. 

28 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Host Response


Immunity against F. tularensis has been studied by evaluating the response of


humans and mice to live vaccine strain (LVS). In humans, LVS vaccination or 


F. tularensis infection induces the proliferation of bacterium-specific T cells and the


production of antibodies, including IgM, IgG, and IgA isotypes. The T-cell


response is directed towards a variety of bacterial proteins, including membrane


and heat-shock proteins; however, no specific epitopes have been identified.


Memory T-cell responses are long-lived and have been documented 10 to 30


years after vaccination. In the mouse, immunity is primarily cellular. In vivo


studies demonstrate that either CD4 or CD8 T cells can mediate resolution of LVS


infections. Antibodies appear to contribute little, if any, to protective immunity.


Passive antibody transfer provides only limited protection against LVS and no


protection against wild type F. tularensis infection in mice.


Vaccines


A live attenuated vaccine derived from an avirulent strain of F. tularensis is available


for laboratory workers. However, the vaccine is not effective for postexposure


immunization because of the incomplete protection it affords and the short


incubation time of inhaled tularemia. Although the correlates of immune protection


are not fully understood, vaccination appears to induce T-cell specific responses to


membrane proteins and to heat shock proteins chaperone-60 (Cpn60) and DnaK.


Long-term studies of survivors infected with tularemia indicate long-lasting


anamnestic responses of T cells to heat shock proteins from F. tularensis.


Therapeutics 

Tularemia has been successfully treated with aminoglycosides, macrolides, 

chloramphenicol, and fluoroquinolones. Recent studies in mice have shown that 

aerosolized liposome-encapsulated ciprofloxacin is more effective in treating 

infection than free aerosolized drug. Knowledge of optimal therapeutic 

approaches for tularemia is incomplete and limited because very few investigators 

are working on this disease. 

29 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Diagnostics 

Rapid diagnostics for tularemia are not widely available, and normal 

microbiological tests are unlikely to result in identification. Culture of F. tularensis 

from patient specimens is the definitive test, but it takes up to several days. PCR, 

antigen detection assays, enzyme-linked immunoassays, and other specialized 

tests that can be used to identify and type tularemia are available at some 

research and reference laboratories. 

Research Resources 

Several resource needs must be addressed to ensure a robust research program 

on F. tularensis. Because most animal species are susceptible to F. tularensis, 

several species have been used in experimental studies. At present, the mouse is 

the most commonly used animal model of disease. Animal evaluations that 

involve infectious strains of F. tularensis will require adequate and secure BSL-3/4 

facilities. In addition, any clinical studies that involve infected patients will require 

appropriate facilities. Attracting new and experienced investigators into F. 

tularensis research also will require appropriate support in terms of facilities, 

reagents, and training. 

Goals 

The following goals are priorities for Biodefense research on tularemia. 

Immediate Research Goals 

•	 Conduct comparative genomic sequencing of selected strains of 

F. tularensis, type A and B, LVS, and F. novicida, and develop genetic 

systems to correlate differences in pathogenesis and virulence. 

• Develop a bacterial repository of Francisella species. 

•	 Characterize responses of F. tularensis to available chemotherapeutics 

in vitro and in animal models of infection and disease. 

•	 Develop new techniques to improve conditions for the culture of the 

microbe and for rapid determination of drug sensitivity profiles. 

•	 Attract scientific researchers with expertise in a diversity of fields (e.g., 

immunology, microbiology, and lipid biochemistry) to the study of tularemia. 

•	 Develop rapid, inexpensive, and broad-based clinical diagnostics 

approaches for tularemia. 

30 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Intermediate and Long-Term Research Goals 

•	 Identify and characterize innate and adaptive immune responses that occur 

after initial exposure to F. tularensis. 

• Identify correlates of immunity for F. tularensis. 

•	 Develop antimicrobials and immunotherapies with novel mechanisms of 

action for treatment of tularemia. 

•	 Synthesize active chemical lead compounds in sufficient quantities for 

preclinical pharmacokinetic and pharmacodynamic analysis and studies of 

efficacy in animal models, mechanism of action, and toxicology. 

•	 Identify new F. tularensis vaccine candidates that can prevent or modulate 

infection, both before and after exposure. 

•	 Conduct phase I and II trials of F. tularensis vaccine candidates and 

alternative adjunctive vaccine formulations. 

•	 Establish capacity for development, refinement, and production of 

candidate F. tularensis vaccines. 

31 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

VIRAL HEMORRHAGIC FEVERS


Viral hemorrhagic fevers (VHFs) encompass a group of similar diseases caused by 

four types of viruses: 

•	 Arenaviruses, associated with Argentine, Bolivian, and Venezuelan 

hemorrhagic fevers, Lassa fever, and Sabia-associated hemorrhagic fever 

•	 Bunyaviruses, including Crimean-Congo hemorrhagic fever, Rift Valley 

fever, and Hantavirus infection 

• Filoviruses, comprising Ebola and Marburg hemorrhagic fevers 

•	 Hemorrhagic flaviviruses, including yellow fever, dengue hemorrhagic fever, 

Kyasanur Forest disease, and Omsk hemorrhagic fever. 

These viruses pose a risk from intentional exposure because, with very few 

exceptions, no vaccines or proven treatments exist, and many of the diseases are 

highly fatal. Natural infections occur when people come in contact with rodents or 

insects that are infected or act as vectors. After human infection occurs, some 

VHFs can be transmitted from person to person through close contact or 

contaminated objects, such as syringes and needles. 

Initial symptoms of VHFs are nonspecific and include fever, muscle aches, and 

fatigue. Disease often progresses to bleeding under the skin and from body 

orifices and internal organs, followed by shock, coma, seizures, and nervous 

system malfunction. Symptoms begin between a few days (in Ebola) and several 

weeks after exposure, depending on the particular virus. Mortality also varies 

widely among the diseases, and it often can be quite high. Some of these viruses 

also cause significant morbidity and mortality in domestic animals that are 

economically important. 

Biology of the Microbe 

The natural life cycle of these viruses is multifaceted. For many viruses, the life 

cycle includes one or more reservoir or amplifying hosts and often an arthropod 

(tick or mosquito) vector. However, for some viruses, such as Ebola, the natural 

reservoirs and/or vectors have not been identified. Infections with viruses that 

cause VHF generally occur in areas where the reservoir hosts or vectors live. 

Contact with body fluids, urine, or feces from rodents that serve as a reservoir can 

lead to infection. Infection also can occur when people come into contact with 

32 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

infected animals, such as monkeys. For example, laboratory workers became 

infected with Marburg hemorrhagic fever after handling monkeys infected with the 

Marburg virus. 

The viruses that cause hemorrhagic fevers are all enveloped RNA viruses. Some 

of the viruses have been extensively studied. The Ebola and Marburg viruses 

replicate in the cytoplasm of infected cells, and viruses bud from the cell surface. 

Viral proteins include a type I transmembrane glycoprotein (GP), a nucleocapsid 

protein, a matrix protein, an RNA-dependent RNA polymerase, and three other 

structural proteins. Ebola virus also codes for a nonstructural soluble GP. Soluble 

GP is found in the serum of infected people and may bind antibodies against the 

transmembrane GP. The transmembrane GP, which is located on the surface of 

the virions and infected cells, may play a role in viral entry via receptor-mediated 

endocytosis. Similar envelope and nucleocapsid proteins, as well as viral 

polymerases, have been identified for the bunyaviruses. 

The arenaviruses contain ribosomes acquired from host cells; the sandy (Latin, 

arenosos) appearance of the ribosomes within the viruses gave rise to their name. 

Replication of arenaviruses includes the transcription of viral proteins from 

genomic sense mRNA, a process termed “ambisense.” 

Host Response 

Arenaviruses induce both T- and B-cell responses that seem to provide life-long 

protection from secondary infection in humans. Neutralizing antibodies appear to 

provide good protection against New World arenavirus infections such as Junin. 

Although Old World arenaviruses, such as Lassa fever, induce high-titer IgG and 

IgM production, these responses do not always correlate with protection. 

Virus-specific T and B cells characterize the immune response to Hantavirus, 

which is a bunyavirus. For example, researchers used the Hantaan strain to 

identify at least 13 CD8 T-cell epitopes presented by the HLA-B51 molecule in 

humans. Studies of B-cell responses to the Puumala strain show that the majority 

of early IgG antibodies are directed against the nucleocapsid protein; however, 

other proteins also are recognized. 

33 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Relatively little is known about the human immune response to Ebola, the most 

well-characterized filovirus. Cell-mediated immunity seems to play an important 

role because surviving patients exhibit stronger T-cell responses than do 

nonsurvivors. A recent study of primary immune responses in asymptomatic 

Ebola-infected individuals identified evidence of T-cell activation and production of 

Ebola-specific antibodies of the IgG1 and IgG3 subclasses. Recent studies in 

guinea pigs, mice, and monkeys demonstrate that DNA vaccines encoding Ebola 

GP or nucleoprotein (NP) induce protective immunity. However, a similar DNA 

vaccine was not effective in primates infected with the Marburg virus. Studies to 

determine the protective effects of anti-Ebola hyperimmune sera in humans and 

animals have been inconclusive. For example, the transfer of neutralizing 

monoclonal antibodies completely protected mice from lethal Ebola infection, even 

if administered after a virus challenge. However, other studies showed the 

existence of infectivity-enhancing antibodies that are directed to the viral GP. 

Flaviviruses, such as dengue, elicit strong immune responses that are directed at 

numerous viral proteins as defined by studies in both humans and mice. Human 

B- and T-cell epitopes have been mapped for dengue. Murine B- and T-cell 

epitopes have been defined for several viruses, including dengue and yellow fever. 

Vaccines 

No vaccines exist for VHFs except for a licensed yellow-fever vaccine. Candidate 

experimental vaccines against Argentine hemorrhagic fever and Rift Valley fever 

have advanced to early human trials. Because of the rapid onset of disease with 

many VHFs, these vaccines may not be useful for postexposure prophylaxis. In 

addition, the presence of different serotypes of some viruses, such as Hantavirus 

and dengue, may require development of multivalent vaccines. 

Recent progress has been made in the development of a potential vaccine for 

Ebola virus. Priming with a DNA vaccine, including the genes for the 

transmembrane GP and nucleocapsid protein, followed by a boost with a 

recombinant adenovirus that contains the same genes, protected cynomolgus 

monkeys against a highly pathogenic challenge. A strong cellular response was 

identified in these animals and two correlates of immune protection have been 

identified, which will help guide human studies. 

34 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Therapeutics 

Generally, there are no effective treatments for VHFs other than supportive care. 

Lassa fever has been successfully treated with the antiviral ribavirin, and plasma 

from recovering patients has been used to treat Argentine hemorrhagic fever. 

Ribavirin was shown to be effective against Hantavirus that causes renal disease 

in China. An open-label observational study in the United States of ribavirin as a 

treatment for Hantavirus pulmonary syndrome was inconclusive. Several potential 

therapeutics showed promise against Ebola infection in mouse models but were 

not effective when tested in cynomolgus monkeys. 

Diagnostics 

Clinical diagnosis of VHFs may be difficult because initial symptoms are 

nonspecific. However, ELISA and PCR tests, as well as viral culture, can be used 

to confirm some infections within days of disease onset. These tests have been 

developed for Ebola, Marburg, Lassa, dengue, Rift Valley fever, and Hantaviruses. 

Research Resources 

Animal models exist for some of the VHFs. For example, Ebola infection of 

nonhuman primates, including African green monkeys, baboons, rhesus monkeys, 

and cynomolgus monkeys, leads to disease that is similar to human disease. 

Ebola that has been adapted to grow in mice and guinea pigs provides a smallanimal model. Animal evaluations that involve infectious strains of most VHFs 

require BSL-3/4 facilities. In addition, appropriate facilities will be required for any 

clinical studies that involve infected patients. 

Goals 

The following goals are priorities for Biodefense research on VHFs. 

Immediate Research Goals 

•	 Develop animal models that mimic human disease for studying VHF 

pathogenesis in humans. 

•	 Expand the determination of the correlates of immunity for VHF vaccines 

by using appropriate models of natural infection. 

•	 Advance the development of existing VHF vaccine candidates (e.g., Rift 

Valley fever and Junin fever). 

35 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

•	 Establish capacity for the development, refinement, and production of pilot 

lots of candidate vaccines for VHFs. 

•	 Develop a centralized immunology laboratory for the validation of tests 

required for licensure of priority VHF vaccines. 

• Screen antibodies to evaluate their possible use as immune therapy for VHFs. 

•	 Obtain clinical samples from patients with VHFs to help validate potential 

diagnostics and aid the development of new vaccine therapies. 

• Expand the in vitro and in vivo screening capability for antivirals against VHFs. 

•	 Encourage the exploration of new targets for antiviral therapies against 

VHFs. 

•	 Complete the genomic sequencing of representative members and 

strains of the VHFs, and compare them to detect differences in 

pathogenesis and virulence. 

Intermediate and Long-Term Research Goals 

•	 Perform comparative studies of VHFs to determine shared mechanisms 

of pathogenesis. 

•	 Expand the genomic analysis of VHFs, and conduct structural studies to 

aid the understanding of function. 

•	 Explore the development of genetic systems to understand VHF replication 

and pathogenesis. 

•	 Establish partnerships as appropriate to expand the study of the ecology of 

VHFs in their natural environment. 

•	 Expand the knowledge of host and vector factors and viral proteins that 

contribute to the pathogenesis and transmission of VHFs. 

• Encourage the structure-based design of new antivirals for multiple VHFs. 

•	 Identify targets within the innate and adaptive pathways that may be used 

to modulate infection. 

•	 Expand the preclinical toxicology capability and clinical capacity needed for 

the development of antivirals against VHF. 

36 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

•	 Synthesize lead compounds in sufficient quantities for preclinical 

pharmacokinetics and studies of animal model efficacy, mechanism of 

action, and toxicology. 

• Examine new technologies for the development of safe vaccines. 

• Conduct phase I and II trials with new candidate VHF vaccines. 

• Initiate human clinical trials of an Ebola vaccine. 

•	 Develop and optimize human monoclonal and polyclonal antibodies as 

therapies for priority VHFs. 

• Evaluate the basis of immune potentiation observed in some VHFs. 

37 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

A p p e n d i x A


NIAID Blue Ribbon Panel on Bioterrorism


and Its Implications for Biomedical Research


February 4–5, 2002


Participant List


A-1 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

PARTICIPANT LIST


Stephen S. Arnon, M.D. 

Infant Botulism Treatment and Prevention Program


California Department of Health Services


State of California


Room 506


2151 Berkeley Way


Berkeley, CA 94704-1011


Phone: (510) 540-2646


Fax: (510) 540-3205


Email: sarnon@dhs.ca.gov


Abdu F. Azad, Ph.D., M.P.H. 

Microbiology and Immunology


University of Maryland School of Medicine


655 West Baltimore Street


Baltimore, MD 21201


Phone: (410) 706-3335


Fax: (410) 706-0282


Email: aazad@umaryland.edu


Phil Baker, Ph.D. 

Program Officer, Lyme Disease Program


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3114


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 435-2855


Fax: (301) 402-2508


Email: pbaker@niaid.nih.gov


Jorge Benach, Ph.D. 

Molecular Genetics and Microbiology


SUNY at Stony Brook


5120 Stony Brook


Stony Brook, NY 11794-5120


Phone: (631) 632-4225


Fax: (631) 632-4294


Email: jbenach@notes.cc.sunysb.edu


Bruce Beutler, M.D. 

Immunology


The Scripps Research Institute


Imm-31


10550 North Torrey Pines Road


La Jolla, CA 92037


Phone: (858) 784-8610


Fax: (858) 784-8444


Email: bruce@scripps.edu


Michael J. Buchmeier, Ph.D. 

Neuropharmacology


Division of Virology, MC CVN-8


The Scripps Research Institute


10550 North Torrey Pines Road


La Jolla, CA 92037


Phone: (858) 784-7056


Fax: (858) 784-7369


Email: buchm@scripps.edu


R. Mark Bueller, Ph.D. 

Departments of Molecular, Microbiology and 

Immunology 

Medical School 

Saint Louis University 

1402 South Grand Boulevard 

St. Louis, MO 63104 

Phone: (314) 577-8451 

Fax: (314) 773-3403 

Email: bullerrm@slu.edu 

Arturo Casadevall, M.D., Ph.D. 

Medicine/Infectious Diseases


Albert Einstein College of Medicine


Golding Building


1300 Morris Park Avenue


Bronx, NY 10461


Phone: (718) 430-3665


Fax: (718) 430-8968


Email: casadeva@aecom.yu.edu


A-2 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Gail H. Cassell, Ph.D. 

Vice President


Eli Lilly and Company


Drop Code 0438


Lilly Corporate Center


Indianapolis, IN 46285


Phone: (317) 276-7374


Fax: (317) 276-1743


Email: cassell_gail_h@lilly.com


Robert Chanock, M.D. 

Chief, Laboratory of Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Building 7, Room 100


9000 Rockville Pike


Bethesda, MD 20892


Phone: (301) 496-4205


Email: rc106v@nih.gov


Frank Chisari, M.D. 

Departments of Molecular and Experimental 

Medicine


Division of Experimental Pathology


The Scripps Research Institute


10550 North Torrey Pines Road


La Jolla, CA 92037


Phone: (858) 784-8228


Fax: (858) 784-2160


Email: fchisari@scripps.edu


Purnell Choppin, M.D. 

President Emeritus


Howard Hughes Medical Institute


4000 Jones Bridge Road


Chevy Chase, MD 20815


Phone: (301) 215-8554


Fax: (301) 215-8566


Email: choppinp@hhmi.org


John Collier, Ph.D. 

Department of Microbiology and Molecular Genetics


Harvard Medical School


Alpert 355


200 Longwood Avenue


Boston, MA 02115


Phone: (617) 432-1930


Fax: (617) 432-0115


Email: jcollier@hms.harvard.edu


Robert B. Couch, M.D. 

Director, Distinguished Service Professor


Center for Infection and Immunity Research


Baylor College of Medicine


Room BCMA-205A, MS BCM-280


One Baylor Plaza


Houston, TX 77030-3498


Phone: (713) 798-4474


Fax: (713) 798-7375


Email: rcouch@bcm.kmc.edu


Walla Dempsey, Ph.D. 

Clinical Trials Program Officer


Virology Branch


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3150


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 496-7453


Fax: (301) 402-0659


Email: wdempsey@niaid.nih.gov


Dennis M. Dixon, Ph.D. 

Chief


Bacteriology and Mycology Branch


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3115


6700-B Rockledge Drive


Bethesda, MD 20892-7630


Phone: (301) 435-2858


Fax: (301) 402-2508


Email: dmdixon@niaid.nih.gov


A-3

NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Eric Eisenstadt, Ph.D. 

Defense Advanced Research Projects Agency


3701 North Fairfax Drive


Arlington, VA 22203-1714


Phone: (703) 696-2322


Fax: (703) 696-3999


Email: eeisenstadt@darpa.mil


Karen L. Elkins, Ph.D. 

Laboratory of Mycobacteria 

Division of Bacterial, Allergenic, 

and Parasitic Products


Center for Biologics Evaluation and Research


HFM 431


1401 Rockville Pike


Rockville, MD 20852


Phone: (301) 496-0544


Fax: (301) 402-2776


Email: elkins@cber.fda.gov


Francis A. Ennis, M.D. 

Center for Infectious Disease and Vaccine Research


University of Massachusetts Medical School


55 Lake Avenue North


Worcester, MA 01655


Phone: (508) 856-4182


Fax: (508) 856-4890


Email: francis.ennis@umassmed.edu


Anthony S. Fauci, M.D. 

Director


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Building 31, Room 7A03C


31 Center Drive


Bethesda, MD 20892


Phone: (301) 496-2263


Fax: (301) 496-4409


Email: afauci@niaid.nih.gov


Claire Fraser, Ph.D. 

The Institute for Genomic Research


9712 Medical Center Drive


Rockville, MD 20850


Phone: (301) 838-3504


Fax: (301) 838-0209


Email: cmfraser@tigr.org


Arthur M. Friedlander, M.D. 

U.S. Army Medical Research Institute 

of Infectious Diseases


Department of Defense


1425 Porter Street


Frederick, MD 21702


Phone: (301) 619-7341


Fax: (301) 619-2152


Email: arthur.friedlander@amedd.army.mil


Maria Giovanni, Ph.D. 

Assistant Director for Microbial Genomics and 

Related Technology Development


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3139


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 496-1884


Fax: (301) 480-4528


Email: mgiovanni@niaid.nih.gov


Irene Glowinski, Ph.D. 

Chief


Office of Scientific Coordination 


and Program Operations


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3145


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 435-8357


Fax: (301) 480-4528


Email: iglowinski@niaid.nih.gov


A-4

NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Jesse L. Goodman, M.D., M.P.H. 

Center for Biologics Evaluation and Research


Food and Drug Administration


Suite 200N HFM-6


1401 Rockville Pike


Rockville, MD 20852


Phone: (301) 435-5680


Fax: (301) 827-0440


Email: goodmanj@cber.fda.gov


Harry B. Greenberg, M.D. 

Microbiology and Immunology


Department of Research


Aviron


297 North Bernardo Avenue


Mountain View, CA 94043


Phone: (650) 919-3711


Fax: (650) 919-2426


Email: hgreenberg@aviron.com


Duane J. Gubler, Sc.D. 

Division of Vector-Borne Infectious Diseases


National Center for Infectious Diseases


Centers for Disease Control and Prevention


Foothills Campus


P.O. Box 2087


Fort Collins, CO 80522


Phone: (970) 221-6428


Fax: (970) 266-3502


Email: dgubler@cdc.gov


Philip Hanna, Ph.D. 

Department of Microbiology and Immunology


University of Michigan Medical School


5641 Medical Science II


Box 0620


Ann Arbor, MI 48109


Phone: (734) 615-3707


Fax: (734) 764-3562


Email: pchanna@umich.edu


Barton F. Haynes, M.D. 

Department of Medicine


Duke University Medical Center


Box 3703, DUMC


Durham, NC 27710


Phone: (919) 681-1095


Fax: (919) 681-8992


Email: hayne002@mc.duke.edu


Carole A. Heilman, Ph.D. 

Director


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3142


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: 301-496-1884


Fax: (301) 480-4528


Email: cheilman@niaid.nih.gov


Thomas V. Inglesby, M.D. 

Deputy Director


Center for Civilian Biodefense Strategies


Johns Hopkins University


Suite 830


111 Market Place


Baltimore, MD 21202


Phone: (410) 223-1655


Fax: (410) 223-1665


Email: tvi@jhsph.edu


Peter B. Jahrling, Ph.D. 

U.S. Army Medical Research Institute 

of Infectious Diseases 

1425 Porter Street 

Fort Detrick, MD 21702-5011 

Phone: (301) 619-2772 

Fax: (301) 619-4625 

Email: peter.jahrling@det.amedd.army.mil 

A-5 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Dennis L. Kasper, M.D. 

Channing Laboratory, Department of Medicine


Brigham and Women’s Hospital


181 Longwood Avenue


Boston, MA 02115


Phone: (617) 525-2280


Fax: (617) 525-0080


Email: dennis_kasper@hms.harvard.edu


Edwin Kilbourne, M.D. 

Department of Microbiology and Immunology


New York Medical College


Basic Science Building


Vahalla, NY 10595


Phone: (914) 594-4193


Fax: (914) 594-4176


Email: ekilbourne@snet.net


John Y. Killen, M.D. 

Associate Director for Research Ethics


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Building 10, Room 1C118


9000 Rockville Pike


Bethesda, MD 20892


Phone: (301) 496-2429


Email: jkillen@niaid.nih.gov


Thomas Kindt, Ph.D. 

Director, Division of Intramural Research


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Building 10, Room 4A31


9000 Rockville Pike


Bethesda, MD 20892


Phone: (301) 496-3006


Email: tk9c@nih.gov


Richard D. Klausner, M.D. 

National Academy of Sciences


2101 Constitution Avenue, NW


Washington, DC 20418


Phone: (202) 334-1238


Fax: (202) 334-1565


Email: coconnor@nas.edu


John La Montagne, Ph.D. 

Deputy Director


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Building 31, Room 7A03B


31 Center Drive


Bethesda, MD 20892


Phone: (301) 496-9677


Fax: (301) 496-4409


Email: jlamontagn@niaid.nih.gov


Sarah Landry, M.F.A. 

Health Science Communication Specialist


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3140


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 496-1884


Fax: (301) 480-4528


Email: slandry@niaid.nih.gov


Clifford Lane, M.D. 

Clinical Director


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Building 10, Room 11B09


9000 Rockville Pike


Bethesda, MD 20892


Phone: (301) 496-7196


Email: c117d@nih.gov


Dennis Lang, Ph.D. 

Enteric Diseases Program Officer


Enteric and Hepatic Diseases Branch


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3110


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 496-7051


Fax: (301) 402-1456


Email: dlang@niaid.nih.gov


A-6

NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Catherine A. Laughlin, Ph.D. 

Chief


Virology Branch


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3150


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 496-7453


Fax: (301) 402-0659


Email: claughlin@niaid.nih.gov


Stanley M. Lemon, M.D. 

School of Medicine


University of Texas Medical Branch


301 University Boulevard


Galveston, TX 77555-0133


Phone: (409) 772-4793


Fax: (409) 772-2684


Email: smlemon@utmb.edu


Stephen Leppla, Ph.D. 

Department of Oral Infection and Immunity


Division of Intramural Research


National Institute of Dental and Craniofacial


Research 

National Institutes of Health 

Building 30, Room 304 

30 Convent Drive 

Bethesda, MD 20892 

Phone: (301) 594-2865 

Fax: (301) 402-1064 

Email: leppla@nih.gov 

Stuart B. Levy, M.D. 

Departments of Molecular Biology, Microbiology, 

and Medicine 

Tufts University School of Medicine 

136 Harrison Avenue 

Boston, MA 02111 

Phone: (617) 636-6764 

Fax: (617) 636-0458 

Email: stuart.levy@tufts.edu 

Margaret Liu, M.D. 

Global Health Program


Bill and Melinda Gates Foundation


P.O. Box 23350


Seattle, WA 98102


Phone: (206) 709-3100


Fax: (206) 709-3170


Email: margaretl@gatesfoundation.org


Adel Mahmoud, M.D., Ph.D. 

President


Merck Vaccines


Merck & Company, Inc.


One Merck Drive


Whitehouse Station, NJ 08889


Phone: (908) 423-4316


Fax: (908) 735-1232


Email: adel_mahmoud@merck.com


Harold S. Margolis, M.D. 

Senior Advisor


Smallpox Preparedness and Response


Office of the Director


Centers for Disease Control and Prevention


1600 Clifton Road


Atlanta, GA 30333


Phone: (404) 639-7000


Email: hsm1@cdc.gov


Grant McFadden, Ph.D. 

University of Western Ontario


Microbiology and Immunology


Robarts Research Institute


1400 Western Road


London, Ontario, Canada N6G2V4


Phone: (519) 663-3184


Fax: (519) 663-3847


Email: mcfadden@rri.ca


A-7 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

John McGowan, Ph.D. 

Director, Division of Extramural Research


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 2142


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 496-7291


Email: jm80c@nih.gov


Pamela McInnes, D.D.S 

Deputy Director


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3142


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 496-1884


Fax: (301) 480-4528


Email: pm23m@nih.gov


Michael Meagher, Ph.D. 

Chemical Engineering


University of Nebraska-Lincoln


East Campus


355 FIB


Lincoln, NE 68583-0919


Phone: (402) 472-2342


Fax: (402) 472-1693


Email: mmeagher1@unl.edu


James Meegan, Ph.D. 

Virology Program Officer


Virology Branch


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3137


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 496-7453


Fax: (301) 402-0659


Email: jmeegan@niaid.nih.gov


Gregory Mertz, M.D. 

Division of Infectious Diseases


Internal Medicine


University of New Mexico


915 Camino De Salud


Albuquerque, NM 87131-5271


Phone: (505) 272-5666


Fax: (505) 272-8441


Email: gmertz@salud.unm.edu


Marissa Miller, D.V.M. 

Antimicrobial Resistance Program Officer


Bacteriology and Mycology Branch


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3244


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 402-8643


Fax: (301) 402-2508


Email: mamiller@niaid.nih.gov


Thomas Monath, M.D., FACP 

Vice President


Department of Research and Medical Affairs


Acambis, Inc.


38 Sidney Street


Cambridge, MA 02139


Phone: (617) 494-1339


Fax: (617) 494-0924


Email: thomas.monath@acambis.com


David Morens, M.D. 

Medical Epidemiologist


Virology Branch


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3149


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 496-7453


Fax: (301) 480-1594


Email: dmorens@niaid.nih.gov


A-8 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Bernard Moss, M.D., Ph.D. 

Laboratory of Viral Diseases


Division of Intramural Research


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Building 4, Room 229A, MSC 0445


4 Center Drive


Bethesda, MD 20892-0445


Phone: (301) 496-9869


Fax: (301) 480-1147


Email: bmoss@nih.gov


Richard W. Moyer, Ph.D. 

Molecular Genetics and Microbiology


University of Florida


P.O. Box 100266


Gainesville, FL 32610


Phone: (352) 392-7077


Fax: (352) 846-2042


Email: rmoyer@mgm.ufl.edu


Barbara Mulach, Ph.D. 

Health Science Communication Specialist


Office of the Director


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3266


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 496-1884


Fax: (301) 480-4528


Email: bmulach@niaid.nih.gov


Brian Murphy, M.D. 

Chief, Laboratory of Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Building 7, Room 106


9000 Rockville Pike


Bethesda, MD 20892


Phone: (301) 496-4205


Email: bmurphy@niaid.nih.gov


James Musser, M.D., Ph.D. 

Laboratory of Human Bacterial Pathogenesis


Division of Intramural Research


National Institute of Allergy and Infectious Diseases


National Institutes of Health


903 South 4B Street


Hamilton, MT 59840


Phone: (406) 363-9287


Fax: (406) 363-9394


Email: jmusser@niaid.nih.gov


Gary Nabel, M.D., Ph.D. 

Director, Vaccine Research Center


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Building 40, Room 4502


9000 Rockville Pike


Bethesda, MD 20892


Phone: (301) 496-1852


Email: gnabel@niaid.nih.gov


Bradley A. Perkins, M.D. 

Chief, Meningitis and Special Pathogens Branch


Division of Bacterial and Mycotic Diseases


Centers for Disease Control and Prevention


MSC C09


1600 Clifton Road


Atlanta, GA 30333


Phone: (404) 639-3158


Fax: (404) 639-3059


Email: bap4@cdc.gov


Robert D. Perry, Ph.D. 

Departments of Microbiology, Immunology, 

and Molecular Genetics


University of Kentucky


MS415 Medical Center


800 Rose Street


Lexington, KY 40536-0298


Phone: (859) 323-6341


Fax: (859) 257-8994


Email: rperry@pop.uky.edu


A-9

NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

C.J. Peters, M.D. 

Center for Biodefense


Departments of Pathology, Microbiology, 


and Immunology


University of Texas Medical Branch


301 University Boulevard


Galveston, TX 77555-0609


Phone: (409) 772-0090


Fax: (409) 772-2500


Robert Purcell, M.D. 

Co-Chief, Laboratory of Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Building 7, Room 202


9000 Rockville Pike


Bethesda, MD 20892


Phone: (301) 496-6227


Email: rpurcell@niaid.nih.gov


David A. Relman, M.D. 

Department of Microbiology, Immunology, 

and Medicine


Veterans Affairs


Stanford University


3801 Miranda Avenue


Palo Alto, CA 94304


Phone: (650) 852-3308


Fax: (650) 852-3291


Email: relman@stanford.edu


Bernard Roizman, Sc.D. 

University of Chicago


910 East 58th Street


Chicago, IL 60637


Phone: (773) 702-1898


Fax: (773) 702-1631


Email: bernard@cummings.uchicago.edu


Martin Rosenberg, Ph.D. 

Research and Development


Promega Corporation


2800 Woods Hollow Road


Madison, WI 53711-5399


Phone: (608) 274-4330


Fax: (608) 277-2601


Email: mrosenbe@promega.com


Daniel Rotrosen, M.D. 

Director, Division of Allergy and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 5142


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 496-1886


Fax: (301) 402-2571


Email: drotrosen@niaid.nih.gov


Philip K. Russell, M.D. 

Office of Public Health and Preparedness


Department of Health and Human Services


Suite G36G


200 Independence Avenue


Washington, DC 20201


Phone: (202) 260-0391


Fax: (202) 690-7412


Email: philip.russell@hhs.gov


Polly Sager, Ph.D. 

Director, Office of Scientific Coordination


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 4149


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 402-0755


Fax: (301) 480-4456


Email: psager@niaid.nih.gov


Jeffrey Schlom, Ph.D. 

Center for Cancer Research


Laboratory of Tumor Immunology and Biology


National Cancer Institute


National Institutes of Health


Room 8B09, MSC 1750


10 Center Drive, Building 10


Bethesda, MD 20892-1750


Phone: (301) 496-4343


Fax: (301) 496-2756


Email: js141c@nih.gov


A-10

NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Connie Schmaljohn, Ph.D. 

Department of Molecular Virology


Virology Division


U.S. Army Medical Research Institute 


of Infectious Diseases 

1301 Ditto Avenue 

Fort Detrick, MD 21702 

Phone: (301) 619-4103 

Fax: (301) 619-2439 

Email: connie.schmaljohn@det.ameddarmy.mil 

Gary K. Schoolnik, M.D. 

Department of Medicine


Stanford Medical School


Room 241


Beckman Center


Stanford, CA 94305


Phone: (650) 723-8158


Fax: (650) 723-1399


Email: schoolni@cmgm.stanford.edu


Lance Simpson, Ph.D. 

Occupational and Environmental Health Sciences


Department of Medicine


Jefferson Medical College


Room 314-JAH


1020 Locust Street


Philadelphia, PA 19107


Phone: (215) 955-8381


Fax: (215) 955-2169


Email: lance.simpson@mail.tju.edu


Christine Sizemore, Ph.D. 

Tuberculosis Program Officer


Respiratory Diseases Branch


Division of Microbiology and Infectious Diseases


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 3244


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 496-5305


Fax: (301) 496-8030


Email: csizemore@niaid.nih.gov


Leonard A. Smith, Ph.D. 

Chief


Department of Immunology and Molecular Biology


Division of Toxinology and Aerobiology


United States Army Medical Research Institute of


Infectious Diseases 

1425 Porter Street 

Fort Detrick, MD 21702-5011 

Phone: (301) 619-4238 

Fax: (301) 619-2348 

Email: leonard.smith@amedd.army.mil 

Susan C. Straley, Ph.D. 

Department of Microbiology, Immunology and 

Molecular Genetics 

University of Kentucky 

800 Rose Street 

Lexington, KY 40536-0298 

Phone: (859) 323-6538 

Fax: (859) 257-8994 

Email: scstra01@uky.edu 

James H. Strauss, Ph.D. 

Division of Biology 156-29


California Institute of Technology


1200 East California Boulevard


Pasadena, CA 91125


Phone: (626) 395-4903


Fax: (626) 796-4209


Email: straussj@caltech.edu


Robert B. Tesh, M.D. 

Center for Tropical Diseases


Department of Pathology


University of Texas Medical Branch


301 University Boulevard


Galveston, TX 77555-0609


Phone: (409) 747-2431


Fax: (409) 747-2429


Email: rtesh@utmb.edu


A-11 
NIAID BIODEFENSE RESEARCH A GENDA FOR CDC CATEGORY A A GENTS 

Ed Tramont, M.D. 

Director, Division of AIDS


National Institute of Allergy and Infectious Diseases


National Institutes of Health


Room 4142


6700-B Rockledge Drive


Bethesda, MD 20892


Phone: (301) 496-0545


Email: etramont@niaid.nih.gov


Emil R. Unanue, M.D. 

Department of Pathology and Immunology


Washington University School of Medicine


660 South Euclid Avenue


Box 8118


St. Louis, MO 63110


Phone: (314) 362-7440


Fax: (314) 362-4096


Email: unanue@pathbox.wustl.edu


Richard J. Whitley, M.D. 

Department of Pediatrics


University of Alabama at Birmingham


Suite 616


1600 Seventh Avenue South


Birmingham, AL 35233


Phone: (205) 934-5316


Fax: (205) 934-8559


Email: rwhitley@peds.uab.edu


John A.T. Young, Ph.D. 

Department of Oncology


McArdle Laboratory for Cancer Research


University of Wisconsin-Madison


1400 University Avenue


Madison, WI 53706


Phone: (608) 265-5151


Fax: (608) 262-2824


Email: young@oncology.wisc.edu


A-12

U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES 

National Institutes of Health 

National Institute of Allergy and Infectious Diseases 

NIH Publication No. 03-5308 

February 2002 

http://biodefense.niaid.nih.gov