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Article ; Online: Crowd control: Bacillus anthracis and quorum sensing.

Rollins, Sean M / Schuch, Raymond

2010 Volume 1, Issue 2, Page(s) 57–59

Abstract: In 1965, Dr. Alexander Tomasz identified a critical component of the DNA uptake mechanism used by competent Streptococcus pneumoniae: the pneumococci secrete a polypeptide that induces the expression of proteins to allow foreign DNA to pass through the ... ...

Abstract	In 1965, Dr. Alexander Tomasz identified a critical component of the DNA uptake mechanism used by competent Streptococcus pneumoniae: the pneumococci secrete a polypeptide that induces the expression of proteins to allow foreign DNA to pass through the bacterium's cell wall. This hormone-like substance was the first of numerous "quorum-sensing" factors that have since been identified in many microbial processes. Detailed insights into the molecular mechanisms of quorum-sensing are now emerging, owing largely to studies focusing on the ability of marine organisms like Vibrio fischeri and Vibrio harveyi to produce light at high cell densities. The complexities of bioluminescence induction, and indeed that of an ever increasing group of other quorum-sensing phenotypes, show that such signaling pathways are not just an interesting phenomenon but rather represent a widespread mechanism by which bacterial populations can communicate, coordinate behavior and act in a cooperative manner in the environment.
MeSH term(s)	Bacillus anthracis/genetics ; Bacillus anthracis/physiology ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Gene Expression Regulation, Bacterial ; Quorum Sensing
Chemical Substances	Bacterial Proteins
Language	English
Publishing date	2010-03
Publishing country	United States
Document type	Comment ; Editorial
ZDB-ID	2657572-3
ISSN	2150-5608 ; 2150-5594
ISSN (online)	2150-5608
ISSN	2150-5594
DOI	10.4161/viru.1.2.11051
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Article: Yersinia pestis and the plague.

Rollins, Sarah E / Rollins, Sean M / Ryan, Edward T

American journal of clinical pathology

2001 Volume 119 Suppl, Page(s) S78–85

Abstract: Yersinia pestis is the cause of plague, an illness that may manifest in bubonic, pneumonic, or septicemic form. Plague has killed an estimated 200 million humans throughout history, and plague is endemic in many areas of the world. Approximately 2,000 ... ...

Abstract	Yersinia pestis is the cause of plague, an illness that may manifest in bubonic, pneumonic, or septicemic form. Plague has killed an estimated 200 million humans throughout history, and plague is endemic in many areas of the world. Approximately 2,000 cases of plague are reported each year to the World Health Organization, and concern has been raised about the possible use of Y pestis as an agent of bioterrorism. The genome of Y pestis has been sequenced, including the 3 virulence plasmids, pPst, pLcr, and pFra, and advances have been made in understanding the bacterial pathogenesis of Y pestis infection. Advances also have been made in rapid diagnosis, the understanding of immune responses during plague, and vaccine development.
MeSH term(s)	Bioterrorism ; Disease Outbreaks ; Genome, Bacterial ; Humans ; Plague/immunology ; Plague/microbiology ; Plague/pathology ; Yersinia pestis/genetics ; Yersinia pestis/pathogenicity
Language	English
Publishing date	2001-10-25
Publishing country	England
Document type	Journal Article ; Research Support, U.S. Gov't, P.H.S. ; Review
ZDB-ID	2944-0
ISSN	1943-7722 ; 0002-9173
ISSN (online)	1943-7722
ISSN	0002-9173
DOI	10.1309/DQM9-3R8Q-NQWB-FYU8
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Article ; Online: Robust microarray production of freshly expressed proteins in a human milieu.

Festa, Fernanda / Rollins, Sean M / Vattem, Krishna / Hathaway, Margarita / Lorenz, Phillip / Mendoza, Eliseo A / Yu, Xiaobo / Qiu, Ji / Kilmer, Greg / Jensen, Penny / Webb, Brian / Ryan, Ed T / LaBaer, Joshua

Proteomics. Clinical applications

2013 Volume 7, Issue 5-6, Page(s) 372–377

Abstract: Purpose: In vitro transcription/translation (IVTT) systems are widely used in proteomics. For clinical applications, mammalian systems are preferred for protein folding and activity; however, the level of protein obtained is low. A new system extracted ... ...

Abstract	Purpose: In vitro transcription/translation (IVTT) systems are widely used in proteomics. For clinical applications, mammalian systems are preferred for protein folding and activity; however, the level of protein obtained is low. A new system extracted from human cells (1-Step Human Coupled IVT (HCIVT)) has the potential to overcome this problem and deliver high yields of protein expressed in a human milieu. Experimental design: Western blots and self-assembled protein microarrays were used to test the efficiency of protein synthesis by HCIVT compared to rabbit reticulocyte lysate (RRL). The arrays were also used to measure the immune response obtained from serum of patients exposed to pathogens or vaccine. Results: HCIVT performed better than RRL in all experiments. The yield of protein synthesized in HCIVT is more than ten times higher than RRL, in both Western blot and protein microarrays. Moreover, HCIVT showed a robust lot-to-lot reproducibility. In immune assays, the signals of many antigens were detected only in HCIVT-expressed arrays, mainly due to the reduction in the background signal and the increased levels of protein on the array. Conclusion and clinical relevance: HCIVT is a robust in vitro transcription and translation system that yields high levels of protein produced in a human milieu. It can be used in applications where protein expression in a mammalian system and high yields are needed. The increased immunogenic response of HCIVT-expressed proteins will be critical for biomarker discovery in many diseases, including cancer.
MeSH term(s)	Animals ; Blotting, Western ; Humans ; Protein Array Analysis ; Protein Biosynthesis ; Proteins/analysis ; Proteins/metabolism ; Rabbits ; Reticulocytes/metabolism
Chemical Substances	Proteins
Language	English
Publishing date	2013-05-17
Publishing country	Germany
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2261788-7
ISSN	1862-8354 ; 1862-8346
ISSN (online)	1862-8354
ISSN	1862-8346
DOI	10.1002/prca.201200063
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Article: Transcutaneous Immunization with Clostridium difficile Toxoid A Induces Systemic and Mucosal Immune Responses and Toxin A-Neutralizing Antibodies in Mice

Ghose, Chandrabali / Kalsy, Anuj / Sheikh, Alaullah / Rollenhagen, Julianne / John, Manohar / Young, John / Rollins, Sean M / Qadri, Firdausi / Calderwood, Stephen B / Kelly, Ciaran P / Ryan, Edward T

Infection and immunity. 2007 June, v. 75, no. 6

2007

Abstract: Clostridium difficile is the leading cause of nosocomial infectious diarrhea. C. difficile produces two toxins (A and B), and systemic and mucosal anti-toxin A antibodies prevent or limit C. difficile-associated diarrhea. To evaluate whether ... ...

Abstract	Clostridium difficile is the leading cause of nosocomial infectious diarrhea. C. difficile produces two toxins (A and B), and systemic and mucosal anti-toxin A antibodies prevent or limit C. difficile-associated diarrhea. To evaluate whether transcutaneous immunization with formalin-treated C. difficile toxin A (CDA) induces systemic and mucosal anti-CDA immune responses, we transcutaneously immunized three cohorts of mice with CDA with or without immunoadjuvantative cholera toxin (CT) on days 0, 14, 28, and 42. Mice transcutaneously immunized with CDA and CT developed prominent anti-CDA and anti-CT immunoglobulin G (IgG) and IgA responses in serum and anti-CDA and anti-CT IgA responses in stool. Sera from immunized mice were able to neutralize C. difficile toxin A activity in an in vitro cell culture assay. CDA itself demonstrated adjuvant activity and enhanced both serum and stool anti-CT IgA responses. Our results suggest that transcutaneous immunization with CDA toxoid may be a feasible immunization strategy against C. difficile, an important cause of morbidity and mortality against which current preventative strategies are failing.
Keywords	Clostridium difficile ; antibodies ; blood serum ; cell culture ; cholera toxin ; diarrhea ; immune response ; immunization ; immunoglobulin A ; immunoglobulin G ; mice ; morbidity ; mortality ; neutralization
Language	English
Size	p. 2826-2832.
Publishing place	American Society for Microbiology
Document type	Article
ZDB-ID	218698-6
ISSN	1098-5522 ; 0019-9567
ISSN (online)	1098-5522
ISSN	0019-9567
Database	NAL-Catalogue (AGRICOLA)

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Article: In vivo induced antigen technology (IVIAT).

Rollins, Sean M / Peppercorn, Amanda / Hang, Long / Hillman, Jeffrey D / Calderwood, Stephen B / Handfield, Martin / Ryan, Edward T

Cellular microbiology

2005 Volume 7, Issue 1, Page(s) 1–9

Abstract: In vivo induced antigen technology (IVIAT) is a technique that identifies pathogen antigens that are immunogenic and expressed in vivo during human infection. IVIAT is complementary to other techniques that identify genes and their products expressed in ... ...

Abstract	In vivo induced antigen technology (IVIAT) is a technique that identifies pathogen antigens that are immunogenic and expressed in vivo during human infection. IVIAT is complementary to other techniques that identify genes and their products expressed in vivo. Genes and gene pathways identified by IVIAT may play a role in virulence or pathogenesis during human infection, and may be appropriate for inclusion in therapeutic, vaccine or diagnostic applications.
MeSH term(s)	Antigens/analysis ; Antigens/biosynthesis ; Gene Expression Regulation ; Gene Library ; Humans ; Immunologic Techniques ; Infection/immunology ; Infection/microbiology
Chemical Substances	Antigens
Language	English
Publishing date	2005-01
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, P.H.S. ; Review
ZDB-ID	1468320-9
ISSN	1462-5822 ; 1462-5814
ISSN (online)	1462-5822
ISSN	1462-5814
DOI	10.1111/j.1462-5822.2004.00477.x
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Zs.A 5288: Show issues

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Article ; Online: Identification of immunogenic Salmonella enterica serotype Typhi antigens expressed in chronic biliary carriers of S. Typhi in Kathmandu, Nepal.

PLoS neglected tropical diseases

2013 Volume 7, Issue 8, Page(s) e2335

Abstract: Background: Salmonella enterica serotype Typhi can colonize and persist in the biliary tract of infected individuals, resulting in a state of asymptomatic chronic carriage. Chronic carriers may act as persistent reservoirs of infection within a ... ...

Abstract	Background: Salmonella enterica serotype Typhi can colonize and persist in the biliary tract of infected individuals, resulting in a state of asymptomatic chronic carriage. Chronic carriers may act as persistent reservoirs of infection within a community and may introduce infection to susceptible individuals and new communities. Little is known about the interaction between the host and pathogen in the biliary tract of chronic carriers, and there is currently no reliable diagnostic assay to identify asymptomatic S. Typhi carriage. Methodology/principal findings: To study host-pathogen interactions in the biliary tract during S. Typhi carriage, we applied an immunoscreening technique called in vivo-induced antigen technology (IVIAT), to identify potential biomarkers unique to carriers. IVIAT identifies humorally immunogenic bacterial antigens expressed uniquely in the in vivo environment, and we hypothesized that S. Typhi surviving in the biliary tract of humans may express a distinct antigenic profile. Thirteen S. Typhi antigens that were immunoreactive in carriers, but not in healthy individuals from a typhoid endemic area, were identified. The identified antigens included a number of putative membrane proteins, lipoproteins, and hemolysin-related proteins. YncE (STY1479), an uncharacterized protein with an ATP-binding motif, gave prominent responses in our screen. The response to YncE in patients whose biliary tract contained S. Typhi was compared to responses in patients whose biliary tract did not contain S. Typhi, patients with acute typhoid fever, and healthy controls residing in a typhoid endemic area. Seven of 10 (70%) chronic carriers, 0 of 8 bile culture-negative controls (0%), 0 of 8 healthy Bangladeshis (0%), and 1 of 8 (12.5%) Bangladeshis with acute typhoid fever had detectable anti-YncE IgG in blood. IgA responses were also present. Conclusions/significance: Further evaluation of YncE and other antigens identified by IVIAT could lead to the development of improved diagnostic assays to identify asymptomatic S. Typhi carriers.
MeSH term(s)	Adult ; Antigens, Bacterial/analysis ; Antigens, Bacterial/immunology ; Biliary Tract/microbiology ; Carrier State/microbiology ; Child ; Child, Preschool ; Gene Expression Profiling ; Host-Pathogen Interactions ; Humans ; Nepal ; Salmonella typhi/immunology ; Typhoid Fever/microbiology
Chemical Substances	Antigens, Bacterial
Language	English
Publishing date	2013-08-01
Publishing country	United States
Document type	Journal Article ; Research Support, American Recovery and Reinvestment Act ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2429704-5
ISSN	1935-2735 ; 1935-2727
ISSN (online)	1935-2735
ISSN	1935-2727
DOI	10.1371/journal.pntd.0002335
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Article ; Online: Identification of in vivo-induced bacterial proteins during human infection with Salmonella enterica serotype Paratyphi A.

Clinical and vaccine immunology : CVI

2013 Volume 20, Issue 5, Page(s) 712–719

Abstract: Salmonella enterica serotype Paratyphi A is a human-restricted pathogen and the cause of paratyphoid A fever. Using a high-throughput immunoscreening technique, in vivo-induced antigen technology (IVIAT), we identified 20 immunogenic bacterial proteins ... ...

Abstract	Salmonella enterica serotype Paratyphi A is a human-restricted pathogen and the cause of paratyphoid A fever. Using a high-throughput immunoscreening technique, in vivo-induced antigen technology (IVIAT), we identified 20 immunogenic bacterial proteins expressed in humans who were bacteremic with S. Paratyphi A but not those expressed in S. Paratyphi A grown under standard laboratory conditions. The majority of these proteins have known or potential roles in the pathogenesis of S. enterica. These include proteins implicated in cell adhesion, fimbrial structure, adaptation to atypical conditions, oxidoreductase activity, proteolysis, antimicrobial resistance, and ion transport. Of particular interest among these in vivo-expressed proteins were S. Paratyphi A (SPA)2397, SPA2612, and SPA1604. SPA2397 and SPA2612 are prophage related, and SPA1604 is in Salmonella pathogenicity island 11 (SPI-11). Using real-time quantitative PCR (RT-qPCR), we confirmed increased levels of mRNA expressed by genes identified by IVIAT in a comparison of mRNA levels in organisms in the blood of bacteremic patients to those in in vitro cultures. Comparing convalescent- to acute-phase samples, we also detected a significant increase in the reaction of convalescent-phase antibodies with two proteins identified by IVIAT: SPA2397 and SPA0489. SPA2397 is a phage-related lysozyme, Gp19, and SPA0489 encodes a protein containing NlpC/P60 and cysteine, histidine-dependent amidohydrolase/peptidase (CHAP) domains. In a previous study utilizing a different approach, we found that transcripts for 11 and 7 of the genes identified by IVIAT were detectable in organisms in the blood of humans in Bangladesh who were bacteremic with S. Paratyphi A and Salmonella enterica serovar Typhi, respectively. S. Paratyphi A antigens identified by IVIAT warrant further evaluation for their contributions to pathogenesis and might have diagnostic, therapeutic, or preventive relevance.
MeSH term(s)	Antigens, Bacterial/biosynthesis ; Antigens, Bacterial/blood ; Antigens, Bacterial/immunology ; Bacteremia/immunology ; Bacteremia/microbiology ; Bacterial Proteins/biosynthesis ; Bacterial Proteins/blood ; Bacterial Proteins/immunology ; Bacterial Proteins/isolation & purification ; Humans ; Paratyphoid Fever/diagnosis ; Paratyphoid Fever/immunology ; Paratyphoid Fever/microbiology ; Prophages ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Salmonella paratyphi A/genetics ; Salmonella paratyphi A/immunology ; Salmonella paratyphi A/virology
Chemical Substances	Antigens, Bacterial ; Bacterial Proteins ; RNA, Messenger
Language	English
Publishing date	2013-03-13
Publishing country	United States
Document type	Journal Article ; Research Support, American Recovery and Reinvestment Act ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2221082-9
ISSN	1556-679X ; 1556-6811
ISSN (online)	1556-679X
ISSN	1556-6811
DOI	10.1128/CVI.00054-13
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Article: Transcutaneous immunization with Clostridium difficile toxoid A induces systemic and mucosal immune responses and toxin A-neutralizing antibodies in mice.

Infection and immunity

2007 Volume 75, Issue 6, Page(s) 2826–2832

Abstract	Clostridium difficile is the leading cause of nosocomial infectious diarrhea. C. difficile produces two toxins (A and B), and systemic and mucosal anti-toxin A antibodies prevent or limit C. difficile-associated diarrhea. To evaluate whether transcutaneous immunization with formalin-treated C. difficile toxin A (CDA) induces systemic and mucosal anti-CDA immune responses, we transcutaneously immunized three cohorts of mice with CDA with or without immunoadjuvantative cholera toxin (CT) on days 0, 14, 28, and 42. Mice transcutaneously immunized with CDA and CT developed prominent anti-CDA and anti-CT immunoglobulin G (IgG) and IgA responses in serum and anti-CDA and anti-CT IgA responses in stool. Sera from immunized mice were able to neutralize C. difficile toxin A activity in an in vitro cell culture assay. CDA itself demonstrated adjuvant activity and enhanced both serum and stool anti-CT IgA responses. Our results suggest that transcutaneous immunization with CDA toxoid may be a feasible immunization strategy against C. difficile, an important cause of morbidity and mortality against which current preventative strategies are failing.
MeSH term(s)	Administration, Cutaneous ; Animals ; Antibodies, Bacterial/analysis ; Antibodies, Bacterial/immunology ; Bacterial Toxins/administration & dosage ; Bacterial Toxins/immunology ; Clostridium difficile/chemistry ; Enterotoxins/administration & dosage ; Enterotoxins/immunology ; Immunity, Mucosal ; Immunization ; Mice ; Mucous Membrane/immunology ; Neutralization Tests ; Toxoids/administration & dosage ; Toxoids/immunology
Chemical Substances	Antibodies, Bacterial ; Bacterial Toxins ; Enterotoxins ; Toxoids ; tcdA protein, Clostridium difficile
Language	English
Publishing date	2007-06
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	218698-6
ISSN	1098-5522 ; 0019-9567
ISSN (online)	1098-5522
ISSN	0019-9567
DOI	10.1128/IAI.00127-07
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Article ; Online: Comparative proteomic analysis of the PhoP regulon in Salmonella enterica serovar Typhi versus Typhimurium.

Charles, Richelle C / Harris, Jason B / Chase, Michael R / Lebrun, Lauren M / Sheikh, Alaullah / LaRocque, Regina C / Logvinenko, Tanya / Rollins, Sean M / Tarique, Abdullah / Hohmann, Elizabeth L / Rosenberg, Ian / Krastins, Bryan / Sarracino, David A / Qadri, Firdausi / Calderwood, Stephen B / Ryan, Edward T

PloS one

2009 Volume 4, Issue 9, Page(s) e6994

Abstract: Background: S. Typhi, a human-restricted Salmonella enterica serovar, causes a systemic intracellular infection in humans (typhoid fever). In comparison, S. Typhimurium causes gastroenteritis in humans, but causes a systemic typhoidal illness in mice. ... ...

Abstract	Background: S. Typhi, a human-restricted Salmonella enterica serovar, causes a systemic intracellular infection in humans (typhoid fever). In comparison, S. Typhimurium causes gastroenteritis in humans, but causes a systemic typhoidal illness in mice. The PhoP regulon is a well studied two component (PhoP/Q) coordinately regulated network of genes whose expression is required for intracellular survival of S. enterica. Methodology/principal findings: Using high performance liquid chromatography mass spectrometry (HPLC-MS/MS), we examined the protein expression profiles of three sequenced S. enterica strains: S. Typhimurium LT2, S. Typhi CT18, and S. Typhi Ty2 in PhoP-inducing and non-inducing conditions in vitro and compared these results to profiles of phoP(-)/Q(-) mutants derived from S. Typhimurium LT2 and S. Typhi Ty2. Our analysis identified 53 proteins in S. Typhimurium LT2 and 56 proteins in S. Typhi that were regulated in a PhoP-dependent manner. As expected, many proteins identified in S. Typhi demonstrated concordant differential expression with a homologous protein in S. Typhimurium. However, three proteins (HlyE, STY1499, and CdtB) had no homolog in S. Typhimurium. HlyE is a pore-forming toxin. STY1499 encodes a stably expressed protein of unknown function transcribed in the same operon as HlyE. CdtB is a cytolethal distending toxin associated with DNA damage, cell cycle arrest, and cellular distension. Gene expression studies confirmed up-regulation of mRNA of HlyE, STY1499, and CdtB in S. Typhi in PhoP-inducing conditions. Conclusions/significance: This study is the first protein expression study of the PhoP virulence associated regulon using strains of Salmonella mutant in PhoP, has identified three Typhi-unique proteins (CdtB, HlyE and STY1499) that are not present in the genome of the wide host-range Typhimurium, and includes the first protein expression profiling of a live attenuated bacterial vaccine studied in humans (Ty800).
MeSH term(s)	Animals ; Bacterial Proteins/metabolism ; Bacterial Vaccines/metabolism ; Gene Expression Regulation, Bacterial ; Humans ; Mice ; Models, Biological ; Mutation ; Peptides/chemistry ; Proteomics/methods ; Salmonella typhi/metabolism ; Salmonella typhimurium/metabolism ; Species Specificity ; Virulence/genetics
Chemical Substances	Bacterial Proteins ; Bacterial Vaccines ; Peptides ; PhoP protein, Bacteria (125360-99-8)
Language	English
Publishing date	2009-09-10
Publishing country	United States
Document type	Comparative Study ; Journal Article ; Research Support, N.I.H., Extramural
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0006994
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Article ; Online: Application of in vivo induced antigen technology (IVIAT) to Bacillus anthracis.

Rollins, Sean M / Peppercorn, Amanda / Young, John S / Drysdale, Melissa / Baresch, Andrea / Bikowski, Margaret V / Ashford, David A / Quinn, Conrad P / Handfield, Martin / Hillman, Jeffrey D / Lyons, C Rick / Koehler, Theresa M / Calderwood, Stephen B / Ryan, Edward T

PloS one

2008 Volume 3, Issue 3, Page(s) e1824

Abstract: In vivo induced antigen technology (IVIAT) is an immuno-screening technique that identifies bacterial antigens expressed during infection and not during standard in vitro culturing conditions. We applied IVIAT to Bacillus anthracis and identified PagA, ... ...

Abstract	In vivo induced antigen technology (IVIAT) is an immuno-screening technique that identifies bacterial antigens expressed during infection and not during standard in vitro culturing conditions. We applied IVIAT to Bacillus anthracis and identified PagA, seven members of a N-acetylmuramoyl-L-alanine amidase autolysin family, three P60 family lipoproteins, two transporters, spore cortex lytic protein SleB, a penicillin binding protein, a putative prophage holin, respiratory nitrate reductase NarG, and three proteins of unknown function. Using quantitative real-time PCR comparing RNA isolated from in vitro cultured B. anthracis to RNA isolated from BALB/c mice infected with virulent Ames strain B. anthracis, we confirmed induced expression in vivo for a subset of B. anthracis genes identified by IVIAT, including L-alanine amidases BA3767, BA4073, and amiA (pXO2-42); the bacteriophage holin gene BA4074; and pagA (pXO1-110). The exogenous addition of two purified putative autolysins identified by IVIAT, N-acetylmuramoyl-L-alanine amidases BA0485 and BA2446, to vegetative B. anthracis cell suspensions induced a species-specific change in bacterial morphology and reduction in viable bacterial cells. Many of the proteins identified in our screen are predicted to affect peptidoglycan re-modeling, and our results support significant cell wall structural remodeling activity during B. anthracis infection. Identification of L-alanine amidases with B. anthracis specificity may suggest new potential therapeutic targets.
MeSH term(s)	Animals ; Antigens, Bacterial/immunology ; Bacillus anthracis/genetics ; Bacillus anthracis/immunology ; Gene Expression Profiling ; Macaca mulatta ; Mice ; Mice, Inbred BALB C ; RNA, Messenger/genetics ; Reverse Transcriptase Polymerase Chain Reaction
Chemical Substances	Antigens, Bacterial ; RNA, Messenger
Language	English
Publishing date	2008-03-19
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0001824
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