bluuebunny/crossref_metadata_2025 huggingface dataset:
Safety and Enhanced Immunogenicity of a Hepatitis B Core ParticlePlasmodium falciparumMalaria Vaccine Formulated in Adjuvant Montanide ISA 720 in a Phase I Trial
Authors: Oliveira Giane A. , Wetzel Kristiane , Calvo-Calle J. Mauricio , Nussenzweig Ruth , Schmidt Annette , Birkett Ashley , Dubovsky Filip , Tierney Eveline , Gleiter Christoph H. , Boehmer Gabriele , Luty Adrian J. F. , Ramharter Michael , Thornton George B. , Kremsner Peter G. , Nardin Elizabeth H.
Published: May 2005

Highly purified subunit vaccines require potent adjuvants in order to elicit optimal immune responses. In a previous phase I trial, an alum formulation of ICC-1132, a malaria vaccine candidate comprising hepatitis B core (HBc) virus-like particle containingPlasmodium falciparumcircumsporozoite (CS) protein epitopes, was shown to elicitPlasmodium falciparum-specific antibody and cellular responses. The present study was designed as a single-blind, escalating-dose phase I trial to evaluate the safety and immunogenicity of single intramuscular doses of ICC-1132 formulated in the more potent water-in-oil adjuvant Montanide ISA 720 (ICC-1132/ISA 720). The vaccine was safe and well tolerated, with transient injection site pain as the most frequent complaint. All vaccinees that received either 20 μg or 50 μg of ICC-1132/ISA 720 developed antiimmunogen and anti-HBc antibodies. The majority of volunteers in these two groups developed sporozoite-specific antibodies, predominantly of opsonizing immunoglobulin G subtypes. Peak titers and persistence of parasite-specific antibody following a single injection of the ISA 720 formulated vaccine were comparable to those obtained following two to three immunizations with alum-adsorbed ICC-1132. Peripheral blood mononuclear cells of ICC-1132/ISA 720 vaccinees proliferated and released cytokines (interleukin 2 and gamma interferon) when stimulated with recombinantP. falciparumCS protein, and CS-specific CD4+T-cell lines were established from volunteers with high levels of antibodies to the repeat region. The promising results obtained with a single dose of ICC-1132 formulated in Montanide ISA 720 encourage further clinical development of this malaria vaccine candidate.
Human Immune Response to a Plague Vaccine Comprising Recombinant F1 and V Antigens
Authors: Williamson E. D. , Flick-Smith H. C. , LeButt C. , Rowland C. A. , Jones S. M. , Waters E. L. , Gwyther R. J. , Miller J. , Packer P. J. , Irving M.
Published: May 2005

The human immune response to a new recombinant plague vaccine, comprising recombinant F1 (rF1) and rV antigens, has been assessed during a phase 1 safety and immunogenicity trial in healthy volunteers. All the subjects produced specific immunoglobulin G (IgG) in serum after the priming dose, which peaked in value after the booster dose (day 21), with the exception of one individual in the lowest dose level group, who responded to rF1 only. Three subjects, found to have an anti-rV titer at screening, were excluded from the overall analysis. Human antibody functionality has been assessed by quantification of antibody competing for binding to rV in vitro and also by the transfer of protective immunity in human serum into the naïve mouse. Human and macaque IgG competed for binding to rV in vitro with a mouse monoclonal antibody, previously shown to protect mice against challenge with plague, suggesting that this protective B-cell epitope on rV is conserved between these three species. Total IgG to rV in individuals and the titer of IgG competing for binding to rV correlated significantly at days 21 (
r
= 0.72;
P
< 0.001) and 28 (
r
= 0.82;
P
< 0.001). Passive transfer of protective immunity into mice also correlated significantly with total IgG titer to rF1 plus rV at days 21 (
r
2
= 98.6%;
P
< 0.001) and 28 (
r
2
= 76.8%;
P
< 0.03). However, no significant vaccination-related change in activation of peripheral blood mononuclear cells was detected at any time. Potential serological immune correlates of protection have been investigated, but no trends specific to vaccination could be detected in cellular markers.
Antimicrobial Activity of Murine Lung Cells against
Staphylococcus aureus
Is Increased In Vitro and In Vivo after Elafin Gene Transfer
Authors: McMichael J. W. , Maxwell A. I. , Hayashi K. , Taylor K. , Wallace W. A. , Govan J. R. , Dorin J. R. , Sallenave J.-M.
Published: May 2005

Staphylococcus aureus
is a pathogen often found in pneumonia and sepsis. In the context of the resistance of this organism to conventional antibiotics, an understanding of the regulation of natural endogenous antimicrobial molecules is of paramount importance. Previous studies have shown that both human and mouse airways express a variety of these molecules, including defensins, cathelicidins, and the four-disulfide core protein secretory leukocyte protease inhibitor. We demonstrate here by culturing mouse tracheal epithelial cells at an air-liquid interface that, despite the production of
Defb1
,
Defb14
, and
Defr1
in this system, these cells are unable to clear
S. aureus
when exposed to this respiratory pathogen. Using an adenovirus (Ad)-mediated gene transfer strategy, we show that overexpression of elafin, an anti-elastase/antimicrobial molecule (also a member of the four-disulfide core protein family), dramatically improves the clearance of
S. aureus
. In addition, we also demonstrate that this overexpression is efficient in vivo and that intratracheal instillation of Ad-elafin significantly reduced the lung bacterial load and demonstrates concomitant anti-inflammatory activity by reducing neutrophil numbers and markers of lung inflammation, such as bronchoalveolar lavage levels of tumor necrosis factor and myeloperoxidase. These findings show that an increased antimicrobial activity phenotype is provided by the elafin molecule and have implications for its use in
S. aureus
-associated local and systemic infections.
Bordetella bronchisepticaAdherence to Cilia Is Mediated by Multiple Adhesin Factors and Blocked by Surfactant Protein A
Authors: Edwards Jessica A. , Groathouse Nathan A. , Boitano Scott
Published: May 2005

In the virulent state (Bvg+),Bordetella bronchisepticaexpresses adhesins and toxins that mediate adherence to the upper airway epithelium, an essential early step in pathogenesis. In this study, we used a rabbit tracheal epithelial cell binding assay to test how specific host or pathogen factors contribute to ciliary binding. The host antimicrobial agent surfactant protein A (SP-A) effectively reduced ciliary binding by Bvg+B. bronchiseptica. To evaluate the relative contributions of bacterial adhesins and toxins to ciliary binding, we used mutant strains ofB. bronchisepticain the binding assay. When compared to Bvg+or Bvg−phase-lockedB. bronchisepticastrains, single-knockout strains lacking one of the known adhesins (filamentous hemagglutinin, pertactin, or fimbriae) displayed an intermediate ciliary binding capacity throughout the coincubation. AB. bronchisepticastrain deficient in adenylate cyclase-hemolysin toxin also displayed an intermediate level of adherence between Bvg+and Bvg−strains and had the lowest ciliary affinity of any of the Bvg+phase strains tested. AB. bronchisepticastrain that was missing dermonecrotic toxin also displayed intermediate binding; however, this strain displayed ciliary binding significantly higher than most of the adhesin knockouts tested. Taken together, these findings suggest that virulent-stateB. bronchisepticaexpresses multiple adhesins with overlapping contributions to ciliary adhesion and that host production of SP-A can provide innate immunity by blocking bacterial adherence to the ciliated epithelium.
Characterization of Fluorescent Chimeras of Cholera Toxin and
Escherichia coli
Heat-Labile Enterotoxins Produced by Use of the Twin Arginine Translocation System
Authors: Tinker Juliette K. , Erbe Jarrod L. , Holmes Randall K.
Published: May 2005

Cholera toxin (CT) is an AB
5
toxin responsible for the profuse secretory diarrhea resulting from
Vibrio cholerae
infection. CT consists of a pentameric, receptor-binding B subunit (CTB) and a monomeric A subunit (CTA) that has latent enzymatic activity. In addition to its enterotoxicity, CT has potent mucosal adjuvant activity and can also function as a carrier molecule with many potential applications in cell biology. In earlier studies, the toxic CTA
1
domain was replaced by several other antigenic protein domains to produce holotoxin-like chimeras for use as potential mucosal vaccines. In the present study we utilized the twin arginine translocation (
tat
) system to produce fluorescent CT chimeras, as well as fluorescent chimeras of
Escherichia coli
heat-labile toxins LTI and LTIIb. Fusion proteins containing either green fluorescent protein (GFP) or monomeric red fluorescent protein (mRFP) and the A
2
domain of CT, LTI, or LTIIb were transported to the periplasm of
E. coli
by the
tat
system, and the corresponding B polypeptides of CT, LTI, and LTIIb were transported to the periplasm by the
sec
system. The fluorescent fusion proteins were shown to assemble spontaneously and efficiently with the corresponding B polypeptides in the periplasm to form chimeric holotoxin-like molecules, and these chimeras bound to and entered cultured cells in a manner similar to native CT, LTI, or LTIIb. The GFP and mRFP derivatives of CT, LT, and LTIIb developed here are useful tools for studies on the cell biology of trafficking of the CT/LT family of bacterial enterotoxins. In addition, these constructs provide proof in principle for the development of novel chimeric CT-like or LT-like vaccine candidates containing CTA
2
fusion proteins that cannot be delivered to the periplasm of
E. coli
by use of the
sec
secretion pathway.
Infection of Newborn Piglets with
Bordetella pertussis
: a New Model for Pertussis
Authors: Elahi S. , Brownlie R. , Korzeniowski J. , Buchanan R. , O'Connor B. , Peppler M. S. , Halperin S. A. , Lee S. F. , Babiuk L. A. , Gerdts V.
Published: May 2005

Bordetella pertussis
is the causative agent of pertussis or whooping cough. This bacterium is a human pathogen that under experimental conditions also infects selected rodents and primates. Here, we show for the first time that newborn piglets can be infected with
B. pertussis
when it is delivered intrapulmonarily. Infected piglets displayed fever and respiratory symptoms, such as nasal discharge, nonparoxysmal coughing, and breathing difficulties. Eventually, all infected animals developed severe bronchopneumonia, which in some cases was combined with a fibrinous pleuritits. Immunohistochemical staining revealed the presence of large numbers of
B. pertussis
cells within airways, adhering to the epithelial lining or phagocytosed by macrophages and neutrophils. Viable bacteria were reisolated from bronchoalveolar lavages and lung lesions for more than 10 days postinfection. The systemic presence of pertussis toxin was shown by hypoglycemia, lymphocytosis, and induction of a clustered pattern of CHO cells by serum and bronchoalveolar lavage samples. Thus, a large-animal model for pertussis was developed, which should complement existing rodent models for identifying the immune responses relevant to the design of new vaccines. In particular, this model should help researchers analyze the roles of both maternal and mucosal immunity in disease protection against pertussis and should ultimately assist in the design of new vaccines for early life protection.
Comparative Secretome Analyses of Three
Bacillus anthracis
Strains with Variant Plasmid Contents
Authors: Lamonica Janine M. , Wagner MaryAnn , Eschenbrenner Michel , Williams Leanne E. , Miller Tabbi L. , Patra Guy , DelVecchio Vito G.
Published: May 2005

Bacillus anthracis
, the causative agent of anthrax, secretes numerous proteins into the extracellular environment during infection. A comparative proteomic approach was employed to elucidate the differences among the extracellular proteomes (secretomes) of three isogenic strains of
B. anthracis
that differed solely in their plasmid contents. The strains utilized were the wild-type virulent
B. anthracis
RA3 (pXO1
+
pXO2
+
) and its two nonpathogenic derivative strains: the toxigenic, nonencapsulated RA3R (pXO1
+
pXO2
−
) and the totally cured, nontoxigenic, nonencapsulated RA3:00 (pXO1
−
pXO2
−
). Comparative proteomics using two-dimensional gel electrophoresis followed by computer-assisted gel image analysis was performed to reveal unique, up-regulated, or down-regulated secretome proteins among the strains. In total, 57 protein spots, representing 26 different proteins encoded on the chromosome or pXO1, were identified by peptide mass fingerprinting. S-layer-derived proteins, such as Sap and EA1, were most frequently observed. Many sporulation-associated enzymes were found to be overexpressed in strains containing pXO1
+
. This study also provides evidence that pXO2 is necessary for the maximal expression of the pXO1-encoded toxins lethal factor (LF), edema factor (EF), and protective antigen (PA). Several newly identified putative virulence factors were observed; these include enolase, a high-affinity zinc uptake transporter, the peroxide stress-related alkyl hydroperoxide reductase, isocitrate lyase, and the cell surface protein A.
Characterization of CD8+T Cells and Microenvironment in Oral Lesions of Human Immunodeficiency Virus-Infected Persons with Oropharyngeal Candidiasis
Authors: McNulty Kelly M. , Plianrungsi Jananya , Leigh Janet E. , Mercante Donald , Fidel Paul L.
Published: May 2005

Oropharyngeal candidiasis (OPC), the most common oral infection in human immunodeficiency virus-positive persons, correlates with reduced blood CD4+T cells. In those with OPC, CD8+T cells accumulate at the lamina propria-epithelium interface at a distance from the organism at the outer epithelium. The present study aimed to characterize the tissue-associated CD8+T cells and tissue microenvironment in both OPC+and OPC−persons. The results show that the majority of CD8+T cells possess the αβ T-cell receptor, the thymus-derived αβ CD8 antigen heterodimer, and similar levels of the α4β7, α4β1, and αeβ7homing receptors. Studies to evaluate the tissue microenvironment showed that in OPC+persons, the adhesion molecule for T cells to enter mucosa, mucosal addressin cell adhesion molecule, is significantly increased, whereas E-cadherin, which allows T cells to migrate through mucosa, is significantly decreased compared to OPC−persons. These results continue to support a role for CD8+T cells against OPC under conditions of reduced numbers of CD4+T cells, with susceptibility to infection potentially associated with a dysfunction in mucosal CD8+T-cell migration by reduced tissue-associated E-cadherin.
Differential Effects of Total and Partial Neutralization of Tumor Necrosis Factor on Cell-Mediated Immunity toMycobacterium bovisBCG Infection
Authors: Guler Reto , Olleros Maria L. , Vesin Dominique , Parapanov Roumen , Garcia Irene
Published: May 2005

The effects of total and partial inhibition of tumor necrosis factor (TNF) on sensitivityto Mycobacterium bovisBCG infection were investigated by using transgenic mice in which hepatocytes produced different amounts of human soluble TNF receptor 1 (sTNFR1) fused to the Fc fragment of human immunoglobulin G3 that could be detected in the serum. Transgenic mice expressing high serum levels of sTNFR1, neutralizing all circulating TNF, failed to develop differentiated granulomas and bactericidal mechanisms, and they succumbed to BCG infection. sTNFR1 transgenic mice did not activate BCG-induced Th1-type cytokines early in infection, but uncontrolled cytokine release was found late in infection. In this work we also evaluated the effect of partial inhibition of TNF on resistance to BCG infection. Transgenic mice expressing low levels of sTNFR1 were protected against BCG infection, and they developed increased bactericidal mechanisms, such as enhanced inducible nitric oxide synthase activity, increased macrophage activation, and showed higher numbers of liver granulomas early in infection compared to their negative littermates. Our data suggest that while total inhibition of TNF prevented BCG-induced cell-mediated immune responses, partial inhibition of TNF could contribute to macrophage activation, induction of bactericidal mechanisms, and granuloma formation in the early phase of BCG infection.
Phase 1 Clinical Trial of Apical Membrane Antigen 1: an Asexual Blood-Stage Vaccine for
Plasmodium falciparum
Malaria
Authors: Malkin Elissa M. , Diemert David J. , McArthur Julie H. , Perreault John R. , Miles Aaron P. , Giersing Birgitte K. , Mullen Gregory E. , Orcutt Andrew , Muratova Olga , Awkal May , Zhou Hong , Wang Jin , Stowers Anthony , Long Carole A. , Mahanty Siddhartha , Miller Louis H. , Saul Allan , Durbin Anna P.
Published: May 2005

Apical membrane antigen 1 (AMA1), a polymorphic merozoite surface protein, is a leading blood-stage malaria vaccine candidate. A phase 1 trial was conducted with 30 malaria-naïve volunteers to assess the safety and immunogenicity of the AMA1-C1 malaria vaccine. AMA1-C1 contains an equal mixture of recombinant proteins based on sequences from the FVO and 3D7 clones of
Plasmodium falciparum
. The proteins were expressed in
Pichia pastoris
and adsorbed on Alhydrogel. Ten volunteers in each of three dose groups (5 μg, 20 μg, and 80 μg) were vaccinated in an open-label study at 0, 28, and 180 days. The vaccine was well tolerated, with pain at the injection site being the most commonly observed reaction. Anti-AMA1 immunoglobulin G (IgG) was detected by enzyme-linked immunosorbent assay (ELISA) in 15/28 (54%) volunteers after the second immunization and in 23/25 (92%) after the third immunization, with equal reactivity to both AMA1-FVO and AMA1-3D7 vaccine components. A significant dose-response relationship between antigen dose and antibody response by ELISA was observed, and the antibodies were predominantly of the IgG1 isotype. Confocal microscopic evaluation of sera from vaccinated volunteers demonstrated reactivity with
P. falciparum
schizonts in a pattern similar to native parasite AMA1. Antigen-specific in vitro inhibition of both FVO and 3D7 parasites was achieved with IgG purified from sera of vaccinees, demonstrating biological activity of the antibodies. To our knowledge, this is the first AMA1 vaccine candidate to elicit functional immune responses in malaria-naïve humans, and our results support the further development of this vaccine.
Strain-Specific Effects of Probiotics on Gut Barrier Integrity following Hemorrhagic Shock
Authors: Luyer Misha D. , Buurman Wim A. , Hadfoune M'hamed , Speelmans Gea , Knol Jan , Jacobs Jan A. , Dejong Cornelis H. C. , Vriesema Aldwin J. M. , Greve Jan Willem M.
Published: May 2005

Probiotic therapy modulates the composition of the intestinal flora and inhibits the inflammatory response. These properties may be of benefit in the preservation of gut barrier integrity after injury or stress. In this study, we examined the effect of two
Lactobacillus
strains selected for their pathogen exclusion properties on intestinal barrier integrity following hemorrhagic shock. Additionally, the responsiveness of the macrophage cell line RAW 264.7 to combined exposure to
Lactobacillus
DNA or oligodeoxynucleotides containing CpG motifs (CpG-ODN) and endotoxin was assessed by measuring tumor necrosis factor alpha (TNF-α) release. Rats were administered lactobacilli (5 × 10
9
CFU) or vehicle for 7 days and were subjected subsequently to hemorrhagic shock by withdrawal of 2.1 ml blood/100 g tissue. Levels of plasma endotoxin, bacterial translocation to distant organs, and filamentous actin (F-actin) in the ileum were determined 24 h later. Rats treated with
Lactobacillus rhamnosus
showed reduced levels of plasma endotoxin (8 ± 2 pg/ml versus 24 ± 4 pg/ml;
P
= 0.01), bacterial translocation (2 CFU/gram versus 369 CFU/gram;
P
< 0.01), and disruption of F-actin distribution following hemorrhagic shock compared with nontreated control rats. In contrast, pretreatment with
Lactobacillus fermentum
had no substantial effect on gut barrier integrity. Interestingly, DNA preparations from both lactobacilli reduced endotoxin-induced TNF-α release dose dependently, whereas CpG-ODN increased TNF-α release. In conclusion, the pathogen exclusion properties of both
Lactobacillus
strains and the reduction of endotoxin-induced inflammation by their DNA in vitro are not prerequisites for a beneficial effect of probiotic therapy on gut barrier function following hemorrhagic shock. Although pretreatment with
Lactobacillus
spp. may be useful to preserve gut barrier integrity following severe hypotension, a thorough assessment of specific strains seems to be essential.
EnhancedPseudomonas aeruginosaBiofilm Development Mediated by Human Neutrophils
Authors: Walker Travis S. , Tomlin Kerry L. , Worthen G. Scott , Poch Katie R. , Lieber Jonathan G. , Saavedra Milene T. , Fessler Michael B. , Malcolm Kenneth C. , Vasil Michael L. , Nick Jerry A.
Published: May 2005

Cystic fibrosis (CF) lung disease features persistent neutrophil accumulation to the airways from the time of infancy. CF children are frequently exposed toPseudomonas aeruginosa, and by adulthood, 80% of CF patients are chronically infected. The formation of biofilms is a particularly important phenotypic characteristic ofP. aeruginosathat allows for bacterial survival despite aggressive antibiotic therapy and an exuberant immune response. Here, we show that the presence of neutrophils enhances initialP. aeruginosabiofilm development over a period of 72 h through the formation of polymers comprised of actin and DNA. F-actin was found to be a site of attachment forP. aeruginosa. These actin and DNA polymers are present in CF sputum, and disruption of the polymers dispersed the associatedP. aeruginosacells and reduced biofilm development. These findings demonstrate a potential maladaptation of the primary innate response. When the host fails to eradicate the infection, cellular components from necrotic neutrophils can serve as a biological matrix to facilitateP. aeruginosabiofilm formation.
Opsonized Virulent
Brucella abortus
Replicates within Nonacidic, Endoplasmic Reticulum-Negative, LAMP-1-Positive Phagosomes in Human Monocytes
Authors: Bellaire Bryan H , Roop R. Martin , Cardelli James A.
Published: May 2005

Cells in the
Brucella
spp. are intracellular pathogens that survive and replicate within host monocytes.
Brucella
maintains persistent infections in animals despite the production of high levels of anti-
Brucella
-specific antibodies. To determine the effect of antibody opsonization on the ability of
Brucella
to establish itself within monocytes, the intracellular trafficking of virulent
Brucella abortus
2308 and attenuated
hfq
and
bacA
mutants was followed in the human monocytic cell line THP-1. Early trafficking events of
B. abortus
2308-containing phagosomes (BCP) were indistinguishable from those seen for control particles (heat-killed
B. abortus
2308, live
Escherichia coli
HB101, or latex beads). All phagosomes transiently communicated the early-endosomal compartment and rapidly matured into LAMP-1
+
, cathepsin D
+
, and acidic phagosomes. By 2 h postinfection, however, the number of cathepsin D
+
BCP was significantly lower for live
B. abortus
2308-infected cells than for either
Brucella
mutant strains or control particles.
B. abortus
2308 persisted within these cathepsin D
−
, LAMP-1
+
, and acidic vesicles; however, at the onset of intracellular replication, the numbers of acidic
B. abortus
2308 BCP decreased while remaining cathepsin D
−
and LAMP-1
+
. In contrast to
B. abortus
2308, the isogenic
hfq
and
bacA
mutants remained in acidic, LAMP-1
+
phagosomes and failed to initiate intracellular replication. Notably, markers specific for the host endoplasmic reticulum were absent from the BCPs throughout the course of the infection. Thus, opsonized
B. abortus
in human monocytes survives within phagosomes that remain in the endosomal pathway and replication of virulent
B. abortus
2308 within these vesicles corresponds with an increase in intraphagosomal pH.
Expression of Genes Encoding Innate Host Defense Molecules in Normal Human Monocytes in Response toCandida albicans
Authors: Kim Hee Sup , Choi Eun Hwa , Khan Javed , Roilides Emmanuel , Francesconi Andrea , Kasai Miki , Sein Tin , Schaufele Robert L. , Sakurai Kenichi , Son Chang Gue , Greer Braden T. , Chanock Stephen , Lyman Caron A. , Walsh Thomas J.
Published: May 2005

Little is known about the regulation and coordinated expression of genes involved in the innate host response toCandida albicans. We therefore examined the kinetic profile of gene expression of innate host defense molecules in normal human monocytes infected withC. albicansusing microarray technology. Freshly isolated peripheral blood monocytes from five healthy donors were incubated withC. albicansfor 0 to 18 h in parallel with time-matched uninfected control cells. RNA from monocytes was extracted and amplified for microarray analysis, using a 42,421-gene cDNA chip. Expression of genes encoding proinflammatory cytokines, including tumor necrosis factor alpha, interleukin 1 (IL-1), IL-6, and leukemia inhibitory factor, was markedly enhanced during the first 6 h and coincided with an increase in phagocytosis. Expression of these genes returned to near baseline by 18 h. Genes encoding chemokines, including IL-8; macrophage inflammatory proteins 1, 3, and 4; and monocyte chemoattractant protein 1, also were strongly up-regulated, with peak expression at 4 to 6 h, as were genes encoding chemokine receptors CCR1, CCR5, CCR7, and CXCR5. Expression of genes whose products may protect monocyte viability, such as BCL2-related protein, metallothioneins, CD71, and SOCS3, was up-regulated at 4 to 6 h and remained elevated throughout the 18-h time course. On the other hand, expression of genes encoding T-cell-regulatory molecules (e.g., IL-12, gamma interferon, and transforming growth factor β) was not significantly affected during the 18-h incubation. Moreover, genes encoding IL-15, the IL-13 receptor (IL-13Ra1), and CD14 were suppressed during the 18-h exposure toC. albicans. Thus,C. albicansis a potent inducer of a dynamic cascade of expression of genes whose products are related to the recruitment, activation, and protection of neutrophils and monocytes.
Effects of Prednisolone Treatment on Cytokine Expression in Patients with Leprosy Type 1 Reactions
Authors: Andersson Anna K. , Chaduvula MeherVani , Atkinson Sara E. , Khanolkar-Young Saroj , Jain Suman , Suneetha Lavanya , Suneetha Sujai , Lockwood Diana N. J.
Published: May 2005

Leprosy type 1 reactions (T1R) are due to increased cell-mediated immunity and result in localized tissue damage. The anti-inflammatory drug prednisolone is used for treatment, but there is little good in vivo data on the molecular actions of prednisolone. We investigated the effect of prednisolone treatment on tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), IL-10, and transforming growth factor β1 (TGF-β1) mRNA and protein expression in blood and skin biopsies from 30 patients with T1R in India. After 1 month of prednisolone treatment the sizes of the skin granulomas were reduced, as were the grades of cells positive for TNF-α and IL-10 in skin lesions. Increased production of TGF-β1 was seen in skin lesions after 6 months of prednisolone treatment. Expression of mRNA for TNF-α, IL-1β, and TGF-β1 was reduced, whereas no change in IL-10 mRNA expression was detected during treatment. The circulating cytokine profiles were similar in patients with and without T1R, and prednisolone treatment had no detectable effects on cytokine expression in the blood. The data emphasize the compartmentalization of pathology in T1R and the importance of the immune response in the skin. Clinical improvement and cytokine expression were compared. Surprisingly, patients with improved skin and nerve function and patients with nonimproved skin and nerve function had similar cytokine profiles, suggesting that clinical improvement is not directly mediated by the cytokines studied here. This in vivo well-controlled study of the immunosuppressive effects of prednisolone showed that the drug does not switch off cytokine responses effectively.
Protein Microarray for Profiling Antibody Responses to
Yersinia pestis
Live Vaccine
Authors: Li Bei , Jiang Lingxiao , Song Qifeng , Yang Junxin , Chen Zeliang , Guo Zhaobiao , Zhou Dongsheng , Du Zongmin , Song Yajun , Wang Jin , Wang Hongxia , Yu Shouyi , Wang Jian , Yang Ruifu
Published: May 2005

A protein microarray representing 149
Yersinia pestis
proteins was developed to profile antibody responses in EV76-immunized rabbits. Antibodies to 50 proteins were detected. There are 11 proteins besides F1 and V antigens to which the predominant antibody response occurred, and these proteins show promise for further evaluation as candidates for subunit vaccines and/or diagnostic antigens.
Deletion of the Ferric Uptake Regulator Fur Impairs the In Vitro Growth and Virulence of
Actinobacillus pleuropneumoniae
Authors: Jacobsen Ilse , Gerstenberger Jörg , Gruber Achim D. , Bossé Janine T. , Langford Paul R. , Hennig-Pauka Isabel , Meens Jochen , Gerlach Gerald-F.
Published: May 2005

In order to investigate the role of the ferric uptake regulator Fur in the porcine lung pathogen
Actinobacillus pleuropneumoniae
, we constructed an isogenic in-frame deletion mutant,
A. pleuropneumoniae
Δ
fur
. This mutant showed constitutive expression of transferrin-binding proteins, growth deficiencies in vitro, and reduced virulence in an aerosol infection model.
Endogenous Interleukin-6 Plays a Crucial Protective Role in Streptococcal Toxic Shock Syndrome via Suppression of Tumor Necrosis Factor Alpha Production
Authors: Diao Hongyan , Kohanawa Masashi
Published: May 2005

During aStreptococcus pyogenesinfection in interleukin-6 (IL-6)-deficient mice, there is elevation of serum tumor necrosis factor alpha (TNF-α) levels, muscular necrosis, and death compared with infection of C57BL/6 mice. Anti-TNF-α monoclonal antibody treatment decreased mortality and muscular necrosis in the infected IL-6-deficient mice. These results suggest that IL-6 plays a crucial protective role via suppression of TNF-α production inS. pyogenesinfection.
Genetic Basis for Conversion of Rough-to-Smooth Colony Morphology in
Actinobacillus actinomycetemcomitans
Authors: Wang Ying , Liu Amy , Chen Casey
Published: May 2005

The basis of the rough-to-smooth conversion of
Actinobacillus actinomycetemcomitans
was examined. Smooth variants often contained mutations at the
flp
promoter region. Replacing the mutated
flp
promoter with the wild-type promoter restored the rough phenotype. The expression level of the
flp
promoter was ∼100-fold lower in smooth than in rough strains. Mutations of the
flp
promoter are a cause of the rough-to-smooth conversion.
Responses of
Mycobacterium tuberculosis
to Growth in the Mouse Lung
Authors: Dubnau Eugenie , Chan John , Mohan V. P. , Smith Issar
Published: May 2005

Using a promoter trap, we have identified 56
Mycobacterium tuberculosis
genes preferentially expressed in the mouse lung. Quantitative real-time PCR showed that RNA levels of several genes were higher from bacteria growing in mouse lungs than from broth cultures. These results support the current hypothesis that
Mycobacterium tuberculosis
utilizes fatty acids as a carbon source in the mouse lung.
Genetic and Functional Analyses of the
Actinobacillus
actinomycetemcomitans
AfeABCD Siderophore-Independent Iron Acquisition System
Authors: Rhodes Eric R. , Tomaras Andrew P. , McGillivary Glen , Connerly Pamela L. , Actis Luis A.
Published: May 2005

The
Actinobacillus
actinomycetemcomitans
afeABCD
iron transport system, the expression of which is controlled by iron and Fur, was identified in three different isolates. The protein products of this locus are related to bacterial ABC transporters involved in metal transport. Transformation of the
Escherichia
coli
1017 iron acquisition mutant with a plasmid harboring
afeABCD
promoted cell growth under iron-chelated conditions. However, insertion disruption of each of the
afeABCD
coding regions abolished this growth-relieving effect. The replacement of the parental
afeA
allele with the derivative
afeA
::EZ::TN drastically reduced the ability of
A. actinomycetemcomitans
cells to grow under iron-chelated conditions.
Effect of Anaerobiosis and Nitrate on Gene Expression in
Pseudomonas aeruginosa
Authors: Filiatrault M. J. , Wagner V. E. , Bushnell D. , Haidaris C. G. , Iglewski B. H. , Passador L.
Published: May 2005

DNA microarrays were used to examine the transcriptional response of
Pseudomonas aeruginosa
to anaerobiosis and nitrate. In response to anaerobic growth, 691 transcripts were differentially expressed. Comparisons of
P. aeruginosa
grown aerobically in the presence or the absence of nitrate showed differential expression of greater than 900 transcripts.
Involvement of Sortase Anchoring of Cell Wall Proteins in Biofilm Formation by
Streptococcus
mutans
Authors: Lévesque Céline M. , Voronejskaia Elena , Huang Yi-Chen Cathy , Mair Richard W. , Ellen Richard P. , Cvitkovitch Dennis G.
Published: May 2005

Streptococcus
mutans
is one of the best-known biofilm-forming organisms associated with humans. We investigated the role of the sortase gene (
srtA
) in monospecies biofilm formation and observed that inactivation of
srtA
caused a decrease in biofilm formation. Genes encoding three putative sortase-dependent proteins were also found to be up-regulated in biofilms versus planktonic cells and mutations in these genes resulted in reduced biofilm biomass.
Expression Cloning of Gamma Interferon-Inducing Antigens ofMycobacteriumaviumsubsp.paratuberculosis
Authors: Nagata Reiko , Muneta Yoshihiro , Yoshihara Kazuhiro , Yokomizo Yuichi , Mori Yasuyuki
Published: May 2005

Three recombinant proteins, Map10, Map39, and Map41, produced based on nucleotide sequences obtained from the screening ofMycobacteriumaviumsubsp.paratuberculosisgenomic library expressed inEscherichiacolisignificantly elicited gamma interferon production in peripheral blood mononuclear cells from infected cattle. Two of these proteins were members of the PPE protein family.
Macrophage Migration Inhibitory Factor Reduces the Growth of VirulentMycobacterium tuberculosisin Human Macrophages
Authors: Oddo Mauro , Calandra Thierry , Bucala Richard , Meylan Pascal R. A.
Published: May 2005

Macrophage migration inhibitory factor (MIF) is a key mediator of the innate immune system and plays a crucial role in the host response to bacterial infections. Its role in immunity to intracellular pathogens has not been well studied. Here, we show that MIF released by infected human macrophages inhibits the growth of virulentMycobacterium tuberculosis.
Recombinant Cysteine Proteinase fromLeishmania (Leishmania) chagasiImplicated in Human and Dog T-Cell Responses
Authors: da Costa Pinheiro Paulo Henrique , de Souza Dias Suzana , Eulálio Kelsen Dantas , Mendonça Ivete L. , Katz Simone , Barbiéri Clara Lúcia
Published: May 2005

High in vitro lymphoproliferative responses were induced in humans and dogs by a recombinantLeishmania(Leishmania)chagasicysteine proteinase, with secretion of IFN-γ in asymptomatic subjects or of IFN-γ, interleukin 4 (IL-4), and IL-10 in oligosymptomatic subjects. In contrast, responses of symptomatic patients and dogs were lower, with production of IL-4 and IL-10.
Entamoeba histolytica
-Secreted Products Degrade Colonic Mucin Oligosaccharides
Authors: Moncada Darcy , Keller Kathy , Chadee Kris
Published: May 2005

Degradation of the mucus layer by
Entamoeba histolytica
is a prerequisite for invasion of the colonic mucosa. In this study, we demonstrate that amoeba-secreted products degrade
3
H-labeled and native colonic mucin oligosaccharides independently of proteolytic activity. We conclude that
E. histolytica
degrades mucin oligosaccharides, which may facilitate parasite invasion of the colon.
Genomic Comparison of
cag
Pathogenicity Island (PAI)-Positive and -Negative
Helicobacter pylori
Strains: Identification of Novel Markers for
cag
PAI-Positive Strains
Authors: Terry Courtney E. , McGinnis Lisa M. , Madigan Katharine C. , Cao Ping , Cover Timothy L. , Liechti George W. , Peek Richard M. , Forsyth Mark H.
Published: May 2005

In an analysis of
Helicobacter pylori
genomic DNA by macroarray methodology, genomic DNA from a panel of
cag
pathogenicity island (PAI)-negative
H. pylori
clinical isolates failed to hybridize with 27 genes located outside the
cag
PAI in a
cag
PAI-positive reference strain. PCR analyses confirmed that HP0217 (encoding a lipopolysaccharide biosynthetic protein) and HP1079 (encoding a protein of unknown function) were present significantly more frequently in
cagA
-positive strains than in
cagA
-negative strains. A low G+C content of these two genes suggests they were acquired by horizontal transfer events.
Combining Suppression Subtractive Hybridization and Microarrays To Map the Intraspecies Phylogeny of
Flavobacterium
psychrophilum
Authors: Soule Marilyn , Cain Kenneth , LaFrentz Stacey , Call Douglas R.
Published: May 2005

Reciprocal subtractive libraries were prepared for two strains of
Flavobacterium
psychrophilum
, one virulent and the other avirulent in a trout challenge model. Unique clones were sequenced and their distribution assessed among 34 strains. The analysis showed that
F. psychrophilum
is composed of two genetic lineages, possibly reflecting host specificity.
Contribution of Nitric Oxide to CpG-Mediated Protection againstListeria monocytogenes
Authors: Ito Shuichi , Ishii Ken J. , Ihata Atsushi , Klinman Dennis M.
Published: May 2005

Immunostimulatory CpG oligodeoxynucleotides (ODN) improve host resistance to listeriae. CpG ODN trigger immune cells to produce gamma interferon and “prime” host cells to secrete nitric oxide in response to bacterial exposure. CpG treatment does not protect inducible nitric oxide synthase 2 knockout mice, indicating that NO is critical to CpG-mediated protection against listeriae.
A
rot
Mutation Restores Parental Virulence to an
agr
-Null
Staphylococcus aureus
Strain in a Rabbit Model of Endocarditis
Authors: McNamara Peter J. , Bayer Arnold S.
Published: May 2005

Mutations in
rot
restore in vitro toxin production to
agr
-negative strains of
Staphylococcus aureus
. We show that a
rot
mutation returns wild-type virulence to an
agr
mutant, as measured in experimental endocarditis infections by target organ bacterial counts. Implications of our data are discussed in terms of
agr
antagonist strategies.
Boosting with Poxviruses Enhances
Mycobacterium bovis
BCG Efficacy against Tuberculosis in Guinea Pigs
Authors: Williams A. , Goonetilleke N. P. , McShane H. , Clark Simon O. , Hatch Graham , Gilbert S. C. , Hill A. V. S.
Published: May 2005

Tuberculosis is rising in the developing world due to poor health care, human immunodeficiency virus type 1 infection, and the low protective efficacy of the
Mycobacterium bovis
BCG vaccine. A new vaccination strategy that could protect adults in the developing world from tuberculosis could have a huge impact on public health. We show that BCG boosted by poxviruses expressing antigen 85A induced unprecedented 100% protection of guinea pigs from high-dose aerosol challenge with
Mycobacterium tuberculosis
, suggesting a strategy for enhancing and prolonging the efficacy of BCG.
Immunological Evaluation and Cellular Location Analysis of the TprI Antigen ofTreponema pallidumsubsp.pallidum
Authors: Giacani Lorenzo , Sambri Vittorio , Marangoni Antonella , Cavrini Francesca , Storni Elisa , Donati Manuela , Corona Silvia , Lanzarini Paolo , Cevenini Roberto
Published: May 2005

The TprI antigen ofTreponema pallidumsubsp.pallidumis a putative virulence factor predicted to be located in the outer membrane of the syphilis spirochete. In this study, we analyzed the immune response against TprI and its subunits in sera collected both from rabbits experimentally infected with the Nichols strain and from patients with syphilis, showing a different pattern of reactivity toward the antigen in these two groups of samples. The protective ability of recombinant TprI and its hypothetical outer membrane location were also investigated. Although no rabbit was protected after challenge, immunoelectron microscopy results, to be further investigated, were compatible with the outer membrane location of the antigen.
CD4+T Cells Play a Dominant Role in Protection against New World Leishmaniasis Induced by Vaccination with the P-4 Amastigote Antigen
Authors: Kar Sujata , Metz Christine , McMahon-Pratt Diane
Published: May 2005

Immunodepletion studies of P-4-vaccinated mice indicate that CD4+and not CD8+T cells are critical for protection againstLeishmania pifanoi(Leishmania mexicanacomplex). Although a moderate CD8+T-cell response is elicited by vaccination, CD4+T cells are the dominant responding population in vitro and at the cutaneous site of infection. These protective T cells produce gamma interferon (IFN-γ), macrophage migration inhibitory factor (MIF), and tumor necrosis factor/lymphotoxin (TNF/LT), each of which significantly contributed to intracellular parasite destruction in vitro. These results indicate that a singular CD4+T-cell response (IFN-γ, MIF, and/or LT/TNF) can provide protection against New World cutaneous leishmaniasis.
A slab-on-slab model for the Flims rockslide (Swiss Alps)
Authors: Pollet Nicolas , Cojean Roger , Couture Réjean , Schneider Jean-Luc , Strom Alexander L , Voirin Claire , Wassmer Patrick
Published: May 2005

The Flims rockslide is the largest landslide in the Alps, with an estimated volume of 12 km3. It resulted from a prehistoric high-speed movement of a large limestone mass. Several main factors influenced the mobility of the Flims rockslide: (i) the steep slopes of the Rhine River valley that blocked the spreading of the rock debris out of the limits of Rabiusa and Carreratobel tributary valleys; (ii) the resisting forces taking place at the base of the rockslide by friction and substratum obstacles; and (iii) the rock mass evolving to a granular state, as observed in the deposits, in which coherence of the original rock massif has been preserved. We expect that most of the energy was consumed by impacting on the opposite slope, forcing the rock mass to stop. Lateral parts and some portions of debris, which entered valleys of the right tributaries of the Rhine River, created tongues by rock avalanche motion, indicating transport velocity. These rock masses eroded the valley fill to create a large mixed mass at the toe of the rockslide deposits. Thus, the Flims rock slope movement can be classified as a rockslide to its middle section and as rock avalanches at its lateral margins. A slab-on-slab model is proposed to characterize transformation of the rock mass during transport, with different stages of motion. Beginning as a rockslide, a delaminating process took place to produce a multislab shearing motion. Shearing and fracturing create dilatancy of the sliding rock debris, with spreading constrained by topographic effects. Dynamic disintegration processes explain the production of fine particles and are at the origin of the granular state of the deposits. Lateral sections of the debris mass continued to flow in the absence of topographic constraints.Key words: rockslide, rock avalanche, Flims, disintegration, topographic control.
2004 Fred Beamish Award Lecture  Analytical  materials chemistry on old CDs  Beyond self-assembly
Authors: Ho Cassie , Soolaman Dinah M , Yu Hua-Zhong
Published: May 2005

This review describes a variety of new chemistry that has been established in our research laboratory and by other scientists worldwide related to optical compact disc (CD) technology. In particular, we have discovered that gold substrates from CD-Rs can be used to prepare high-quality, self-assembled monolayers (SAMs) after removing the protective polymer films with concentrated nitric acid. It was proven that these substrates are feasible for electrochemical, spectroscopic, and surface wetting studies. We have also demonstrated that CD-R gold films are ideal micropatterned conductive substrates for the "customized" fabrication of material microstructures. In addition, we have been working on the micropatterning of DNA probes on CD-R substrates, and exploring the possibility of using the CD-R writing/reading system for biochip fabrication and gene analysis.Key words: compact disc-recordable (CD-R), self-assembled monolayers (SAMs), electrochemical analysis, material micro- and (or) nanostructures, biomedical diagnosis.
Browse site selection by snowshoe hares: effects of food supply and predation risk
Authors: Hodges K E , Sinclair A R.E
Published: May 2005

If snowshoe hares (Lepus americanus Erxleben, 1777) change their foraging behaviour through the population cycle as food supply and predation pressure change, these shifts could contribute to their population cycles by affecting survival and reproduction. We examined whether hares change their foraging movements and browse site selection in response to manipulations of food addition and predator reduction during a cyclic low phase. Snowshoe hares on sites with supplemental rabbit chow ate fewer species per site and preferred to browse in slightly denser cover than unfed hares. Differences in foraging behaviour were linked to season and site characteristics. Snowshoe hares moved similar distances and spent similar amounts of time per browse site in the presence and absence of terrestrial predators. Hares protected from predators used slightly more browse sites in thick cover, but this pattern was partially due to differences in availability. The absence of terrestrial predators had little effect on snowshoe hare foraging behaviour; instead, browse distribution patterns explained most of the behavioural variation. Thus, the predicted patterns in response to the manipulations did not occur, and our results challenge the idea that changes in snowshoe hare foraging behaviour contribute to their cyclic dynamics.
Comment on "Identifying spiders through DNA barcodes"
Authors: Prendini Lorenzo
Published: May 2005

R.D.H. Barrett and P.D.N Hebert have demonstrated that it is possible to identify members of a mostly local spider fauna using a short fragment of the mitochondrial gene coding for cytochrome c oxidase I. There are instances where DNA-based identification may be very useful, e.g., in identifying juvenile life stages of groups in which adults are required for morphological identification, or matching morphologically different sexes or life stages when those associations are unknown. DNA-based identification may be the easiest and most cost-effective way, or even the only feasible way, to address some of these questions. However, these are also the least challenging problems in taxonomy, and their solution is unlikely to relieve the "taxonomic impediment". Furthermore, to promote the utility of DNA barcoding as a global identification system, these authors must demonstrate that their approach works for distinguishing all the members of a speciose clade, wherever in the world they occur. Much of diversity occurs allopatrically and neither the study by R.D.H. Barrett and P.D.N. Hebert, nor any other presented to date, even begins to address the feasibility of DNA-based identification at this level of detail.
NEW QUANTUM "az+b" GROUPS
Authors: SOŁTAN PIOTR MIKOŁAJ
Published: May 2005

We construct quantum "az+b" groups for new values of the deformation parameter. Along the way we introduce new special functions and study their analytic properties as well as analyze the commutation relations determined by the choice of parameter.