Mucosal Immunity, Vaccines and Microbiota Interplay in Humans and Animal

Summary

Principal Investigator: Marcelo B Sztein
Abstract: DESCRIPTION (provided by applicant): Vaccines are widely viewed as cost-effective interventions to prevent and control classical endemic and epidemic infectious diseases, as well as to limit the transmission and impact of emerging infections and certain bioterror agents. However, the development of new and improved vaccines against some of these agents is hampered by a lack of information concerning the "true" (i.e., operative) immunological mechanisms underlying the protection elicited by natural infection and by candidate vaccines. This is particularly true with regard to pathogens that enter the host via mucosal surfaces, including the gastrointestinal (Gl) tract. Thus, this proposal is focused on furthering our understanding of the protective immunological mechanisms that can be elicited in the Gl microenvironment of humans. Moreover, because the normal Gl flora (microbiota) is certain to influence the host immune response, we propose to conduct pioneering studies on the interactions between the local intestinal microbiota and the host immune response in humans. We will focus our efforts on in-depth studies of 3 major gram negative bacterial human pathogens including Helicobacter pylori, Salmonella enterica serovar Typhi and Shigella, each primarily affecting a distinct major segment of the Gl tract (i.e., the stomach, ileum and colon, respectively). Because virtually all of the limited information available concerning the determinants of protective mucosal immunity comes from studies in adults, we will also focus some efforts in exploring the responses to oral immunization with the licensed Ty21a typhoid in children and, for the first time, the elderly. To address the complexity of this undertaking, we have assembled a multidisciplinary team consisting of renowned investigators in the fields of innate and adaptive immunity, molecular biology, mucosal biology and physiology, biochemistry, high-throughput technology, microbiology, genomics, protein chemistry and clinical gastroenterology and vaccinology (with extensive experience in performing endoscopies and in conducting vaccine trials). In addition, we propose to develop two novel technologies to broadly advance the study of human immunology, including a human-based approach to study the entire S. Typhi ORFeome to identify CD8+ T cell responses and a peptide conformation constrainment technology and potential mucosal adjuvants to advance H. pylori vaccines. We expect this CCHI to yield much needed information in an area of great importance to human health. RELEVANCE: In spite of the great need for new and improved vaccines against major human pathogens, particularly those that enter the human host through the intestinal mucosa, this field is impeded by insufficient knowledge of the determinants of protective gut immunity. Moreover, little is known concerning the role of the gut microbiota in modulating the immunogenicity of oral vaccines and vice versa. The wealth of data generated by these studies is likely to lead to major advances in mucosal vaccine development in humans. PROJECT 1: PROTECTIVE IMMUNE MECHANISMS TO S. DYSENTERIAE 1 VACCINES IN CYNOMOLGUS MACAQUES AND HUMANS (Sztein, M) PROJECT 1 DESCRIPTION (provided by applicant): Shigella is a global infection that is notorious for disseminating rapidly in certain settings. One serotype, Shigella dysenteriae type 1 (S. dysenteriae 1), can cause devastating pandemics with high case fatality rates and thus it has been classified as a Category B priority pathogen with high potential to be used as a biological weapon. There is no available vaccine for Shigella. The development of effective Shigella vaccines has been hampered by a considerable lack of information of the specific determinants of protective immunity to Shigella infection. Because of the limitations imposed by the risks associated with performing challenge studies with wild type S. dysenteriae 1 in clinical trials to advance vaccine development, a nonhuman primate model is urgently needed. We have already established a challenge model with wild-type S. dysenteriae 1 strain 1617 which, to date, exhibited an attack rate of 100% (6 of 6 cynomolgus macaques challenged with 10e11 cfu intragastrically). Furthermore, we have advanced our understanding of the immune responses elicited following challenge. In this application we propose to continue these studies by addressing the following Specific Aims: (1) evaluate the hypothesis that intragastric immunization with novel attenuated S. dysenteriae 1 mutant strains elicits protection from intragastric challenge with wild type S. dysenteriae 1;(2) evaluate the hypotheses that a defined set of immune responses observed in circulation in cynomolgus immunized with attenuated strains of S. dysenteriae 1 and/or challenged with wild type S. dysenteriae 1 correlate with protection and are representative of those present at effector sites (i.e., mucosal tissues) and secondary lymphoid organs. These translational studies are central to further our understanding of the immunological mechanisms that mediate protection to S. dysenteriae 1 and longevity of the responses to vaccination in humans, (3) To evaluate the effects of immunization of monkeys with attenuated S. dysenteriae 1 strains on the colonic microbiota in stools of monkeys and the impact of the existing microbiota on the observed immune responses and protection from challenge. Finally, we will take advantage of an upcoming trial with the attenuated S. dysenteriae 1 strain CVD 1256 to evaluate the hypothesis that the immune responses observed systemically and locally in humans are similar to those that correlate with protection in cynomolgus macaques (Aims 1 and 2). These studies will provide valuable insights that might accelerate the development of attenuated vaccines for S. dysenteriae 1. RELEVANCE: The overall objective of this project is to develop a safe and effective vaccine for S. dysenteriae 1, a Category B priority pathogen with potential to be used as a biological weapon. Currently, there is no available vaccine for Shigella and limited treatment options for infections with multiple antibiotic resistant strains. Given the shortcomings of available measures to successfully control this infection, and its bioterrorism potential, to develop a S. dysenteriae type 1 vaccine is of great importance.
Funding Period: 2009-06-18 - 2014-05-31
more information: NIH RePORT

Top Publications

  1. pmc Laboratory maintenance of helicobacter species
    Thomas G Blanchard
    Case Western Reserve University, Cleveland, Ohio, USA
    Curr Protoc Microbiol . 2006
  2. pmc Progress and pitfalls in Shigella vaccine research
    Eileen M Barry
    University of Maryland School of Medicine, Center for Vaccine Development, Baltimore, MD 21201, USA
    Nat Rev Gastroenterol Hepatol 10:245-55. 2013
  3. pmc The two-faced T cell epitope: examining the host-microbe interface with JanusMatrix
    Leonard Moise
    Institute for Immunology and Informatics University of Rhode Island Providence, RI, USA EpiVax Inc Providence, RI USA
    Hum Vaccin Immunother 9:1577-86. 2013
  4. pmc Applying mathematical tools to accelerate vaccine development: modeling Shigella immune dynamics
    Courtney L Davis
    Natural Science Division, Pepperdine University, Malibu, California, United States of America
    PLoS ONE 8:e59465. 2013
  5. pmc Impact of oral typhoid vaccination on the human gut microbiota and correlations with s. Typhi-specific immunological responses
    Emiley A Eloe-Fadrosh
    Institute for Genome Sciences, University of Maryland School of Medicine, Baltimore, Maryland, United States of America
    PLoS ONE 8:e62026. 2013
  6. pmc Shigella antigen-specific B memory cells are associated with decreased disease severity in subjects challenged with wild-type Shigella flexneri 2a
    Rezwanul Wahid
    Center for Vaccine Development, Department of Pediatrics, University of Maryland School of Medicine, Baltimore, MD, USA
    Clin Immunol 148:35-43. 2013
  7. pmc Genome sequences of 65 Helicobacter pylori strains isolated from asymptomatic individuals and patients with gastric cancer, peptic ulcer disease, or gastritis
    Thomas G Blanchard
    Department of Pediatrics, University of Maryland, Baltimore, MD, USA
    Pathog Dis 68:39-43. 2013
  8. pmc Differential response of the cynomolgus macaque gut microbiota to Shigella infection
    Anna M Seekatz
    Institute for Genome Sciences, University of Maryland School of Medicine, Baltimore, Maryland, United States of America
    PLoS ONE 8:e64212. 2013
  9. pmc A systematic approach toward stabilization of CagL, a protein antigen from Helicobacter pylori that is a candidate subunit vaccine
    Shyamal P Choudhari
    Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA
    J Pharm Sci 102:2508-19. 2013
  10. pmc Dengue vaccines: recent developments, ongoing challenges and current candidates
    Monica A McArthur
    Department of Pediatrics, University of Maryland, School of Medicine, 685 West Baltimore Street, Room 480, Baltimore, MD 21201, USA
    Expert Rev Vaccines 12:933-53. 2013

Detail Information

Publications29

  1. pmc Laboratory maintenance of helicobacter species
    Thomas G Blanchard
    Case Western Reserve University, Cleveland, Ohio, USA
    Curr Protoc Microbiol . 2006
    ..This unit describes how to culture the most commonly used gastric Helicobacter species, H. pylori, H. mustelae, and H. felis...
  2. pmc Progress and pitfalls in Shigella vaccine research
    Eileen M Barry
    University of Maryland School of Medicine, Center for Vaccine Development, Baltimore, MD 21201, USA
    Nat Rev Gastroenterol Hepatol 10:245-55. 2013
    ..Herein, we review the latest progress in Shigella vaccine development within the framework of persistent obstacles...
  3. pmc The two-faced T cell epitope: examining the host-microbe interface with JanusMatrix
    Leonard Moise
    Institute for Immunology and Informatics University of Rhode Island Providence, RI, USA EpiVax Inc Providence, RI USA
    Hum Vaccin Immunother 9:1577-86. 2013
    ....
  4. pmc Applying mathematical tools to accelerate vaccine development: modeling Shigella immune dynamics
    Courtney L Davis
    Natural Science Division, Pepperdine University, Malibu, California, United States of America
    PLoS ONE 8:e59465. 2013
    ..It introduces mathematical models to the Shigella vaccine development effort and lays a foundation for joint theoretical/experimental/clinical approaches to Shigella vaccine design...
  5. pmc Impact of oral typhoid vaccination on the human gut microbiota and correlations with s. Typhi-specific immunological responses
    Emiley A Eloe-Fadrosh
    Institute for Genome Sciences, University of Maryland School of Medicine, Baltimore, Maryland, United States of America
    PLoS ONE 8:e62026. 2013
    ..These results provide an unprecedented view into the dramatic temporal heterogeneity of both the gut microbiota and host immune responses...
  6. pmc Shigella antigen-specific B memory cells are associated with decreased disease severity in subjects challenged with wild-type Shigella flexneri 2a
    Rezwanul Wahid
    Center for Vaccine Development, Department of Pediatrics, University of Maryland School of Medicine, Baltimore, MD, USA
    Clin Immunol 148:35-43. 2013
    ..Inverse correlations between magnitude of pre-challenge IgG antibodies to LPS and IpaB, as well as IgA IpaB-BM and post-challenge IgA LPS-BM with disease severity suggest a role for antigen-specific BM in protection...
  7. pmc Genome sequences of 65 Helicobacter pylori strains isolated from asymptomatic individuals and patients with gastric cancer, peptic ulcer disease, or gastritis
    Thomas G Blanchard
    Department of Pediatrics, University of Maryland, Baltimore, MD, USA
    Pathog Dis 68:39-43. 2013
    ....
  8. pmc Differential response of the cynomolgus macaque gut microbiota to Shigella infection
    Anna M Seekatz
    Institute for Genome Sciences, University of Maryland School of Medicine, Baltimore, Maryland, United States of America
    PLoS ONE 8:e64212. 2013
    ..These studies highlight the importance of further investigation into the possible protective role of the microbiota against enteric pathogens and consideration of host genetic backgrounds in conducting vaccine studies...
  9. pmc A systematic approach toward stabilization of CagL, a protein antigen from Helicobacter pylori that is a candidate subunit vaccine
    Shyamal P Choudhari
    Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA
    J Pharm Sci 102:2508-19. 2013
    ..Candidate stabilizers were selected to confirm their enhanced stabilizing effect. These analyses will help in the formulation of a stable vaccine against H. pylori...
  10. pmc Dengue vaccines: recent developments, ongoing challenges and current candidates
    Monica A McArthur
    Department of Pediatrics, University of Maryland, School of Medicine, 685 West Baltimore Street, Room 480, Baltimore, MD 21201, USA
    Expert Rev Vaccines 12:933-53. 2013
    ..Here, the recent advances in dengue virus vaccine development are reviewed and the challenges associated with the use of these vaccines as a public health tool are briefly discussed. ..
  11. pmc Effect of wild-type Shigella species and attenuated Shigella vaccine candidates on small intestinal barrier function, antigen trafficking, and cytokine release
    Maria Fiorentino
    Mucosal Immunology and Biology Research Center, Massachusetts General Hospital, Charlestown, Massachusetts, United States of America
    PLoS ONE 9:e85211. 2014
    ..Diarrhea may be a mechanism by which the host attempts to eliminate harmful bacteria and transport them from the small to the large intestine where they invade colonocytes inducing a strong inflammatory response. ..
  12. pmc Live oral Salmonella enterica serovar Typhi vaccines Ty21a and CVD 909 induce opsonophagocytic functional antibodies in humans that cross-react with S. Paratyphi A and S. Paratyphi B
    Rezwanul Wahid
    Center for Vaccine Development, Departments of Pediatrics and Medicine, University of Maryland School of Medicine, Baltimore, Maryland, USA
    Clin Vaccine Immunol 21:427-34. 2014
    ..The CVD 909 study has been registered at ClinicalTrials.gov under registration no. NCT00326443.). ..
  13. pmc B cells modulate mucosal associated invariant T cell immune responses
    Rosângela Salerno-Goncalves
    Department of Pediatrics, Center for Vaccine Development CVD, University of Maryland School of Medicine, Baltimore, MD, USA
    Front Immunol 4:511. 2014
    ..These results provide important insights into the role of B cells as a source of antigen-presenting cells to MAIT cells and the gut immune surveillance of commensal microbiota...
  14. pmc Unexpected heterogeneity of multifunctional T cells in response to superantigen stimulation in humans
    Monica A McArthur
    Center for Vaccine Development, University of Maryland School of Medicine, Baltimore MD 21201, USA
    Clin Immunol 146:140-52. 2013
    ..These results suggest a possible role for IL-17A-producing multifunctional T cells in the pathogenesis of TSS...
  15. pmc The human microbiome: from symbiosis to pathogenesis
    Emiley A Eloe-Fadrosh
    Institute for Genome Sciences, University of Maryland School of Medicine, Baltimore, Maryland 21201, USA
    Annu Rev Med 64:145-63. 2013
    ..In line with these predictions, it is possible that existing conditions may be resolved by altering the microbiota in a positive way...
  16. pmc Helicobacter pylori-induced disruption of monolayer permeability and proinflammatory cytokine secretion in polarized human gastric epithelial cells
    Maria Fiorentino
    University of Maryland, Mucosal Biology Research Center, Baltimore, Maryland, USA
    Infect Immun 81:876-83. 2013
    ..The NCI-N87 cell line provides an excellent model for the in vitro study of H. pylori pathogenesis and the epithelial cell host response to infection...
  17. pmc A challenge model for Shigella dysenteriae 1 in cynomolgus monkeys (Macaca fascicularis)
    Steven T Shipley
    Department of Pathology, University of Maryland School of Medicine, Baltimore, USA
    Comp Med 60:54-61. 2010
    ..We believe that this animal model may provide unique insights into the immunologic mechanisms of protection to S. dysenteriae 1 infection and in advancing development of a vaccine against shigellosis...
  18. pmc Ex Vivo kinetics of early and long-term multifunctional human leukocyte antigen E-specific CD8+ cells in volunteers immunized with the Ty21a typhoid vaccine
    Rosângela Salerno-Goncalves
    Center for Vaccine Development, University of Maryland School of Medicine, Baltimore, MD 21201, USA
    Clin Vaccine Immunol 17:1305-14. 2010
    ..Typhi. Finally, this is the first demonstration, in either animals or humans, of the presence of long-term multifunctional HLA-E-restricted CD8(+) cells after immunization...
  19. pmc Maintenance of Helicobacter pylori cultures in agar stabs
    Jinghua Xu
    Department of Pediatrics, University of Maryland School of Medicine, Baltimore, MD 21201, USA
    Helicobacter 15:477-80. 2010
    ..Agar stab cultures were assessed as a possible means of maintaining viability without the need to passage every 4-7 days...
  20. pmc Oral priming with Salmonella Typhi vaccine strain CVD 909 followed by parenteral boost with the S. Typhi Vi capsular polysaccharide vaccine induces CD27+IgD-S. Typhi-specific IgA and IgG B memory cells in humans
    Rezwanul Wahid
    Center for Vaccine Development, Department of Pediatrics, University of Maryland School of Medicine, Baltimore, MD, USA
    Clin Immunol 138:187-200. 2011
    ..The persistent IgA B(M) responses demonstrate the capacity of oral typhoid vaccines to prime mucosally relevant immune memory...
  21. pmc Immunology of gut mucosal vaccines
    Marcela F Pasetti
    Center for Vaccine Development, University of Maryland School of Medicine, 685 West Baltimore St, Room 480, Baltimore, MD 21201, USA
    Immunol Rev 239:125-48. 2011
    ..This review dissects the immune responses elicited in humans by enteric vaccines...
  22. pmc Antigen-specific IgA B memory cell responses to Shigella antigens elicited in volunteers immunized with live attenuated Shigella flexneri 2a oral vaccine candidates
    J K Simon
    Division of Infectious Diseases and Tropical Pediatrics, Department of Pediatrics, University of Maryland School of Medicine, Baltimore, MD, USA
    Clin Immunol 139:185-92. 2011
    ..These B(M) cell responses are likely play an important role in modulating the magnitude and longevity of the humoral response...
  23. ncbi Laboratory maintenance of Helicobacter species
    Thomas G Blanchard
    Department of Pediatrics, University of Maryland, Baltimore, Maryland, USA
    Curr Protoc Microbiol . 2012
    ..This unit describes how to culture the most commonly used gastric Helicobacter species, H. pylori, H. mustelae, and H. felis...
  24. pmc Live oral typhoid vaccine Ty21a induces cross-reactive humoral immune responses against Salmonella enterica serovar Paratyphi A and S. Paratyphi B in humans
    Rezwanul Wahid
    Department of Pediatrics, Center for Vaccine Development, University of Maryland School of Medicine, Baltimore, Maryland, USA
    Clin Vaccine Immunol 19:825-34. 2012
    ..Whereas cross-reactive humoral immune responses to S. Paratyphi A or B antigens are demonstrable following Ty21a immunization, they cannot explain the efficacy data gleaned from controlled field trials...
  25. ncbi 'Omics' of the mammalian gut--new insights into function
    Regina Lamendella
    Lawrence Berkley National Laboratory, 1 Cyclotron Road, Berkeley, CA 92597, USA
    Curr Opin Biotechnol 23:491-500. 2012
    ..In addition, we address some promises that these techniques hold for future therapeutic and diagnostic applications...
  26. pmc Heterogeneity of multifunctional IL-17A producing S. Typhi-specific CD8+ T cells in volunteers following Ty21a typhoid immunization
    Monica A McArthur
    Center for Vaccine Development, University of Maryland School of Medicine, Baltimore, Maryland, United States of America
    PLoS ONE 7:e38408. 2012
    ....
  27. pmc Diverse phosphorylation patterns of B cell receptor-associated signaling in naïve and memory human B cells revealed by phosphoflow, a powerful technique to study signaling at the single cell level
    Franklin R Toapanta
    Department of Medicine, Center for Vaccine Development, University of Maryland Baltimore, MD, USA
    Front Cell Infect Microbiol 2:128. 2012
    ..Phosphoflow holds great potential to accelerate vaccine development by identifying signaling profiles in good/poor responders...
  28. pmc An outpatient, ambulant-design, controlled human infection model using escalating doses of Salmonella Typhi challenge delivered in sodium bicarbonate solution
    Claire S Waddington
    Oxford Vaccine Group, Department of Paediatrics, University of Oxford
    Clin Infect Dis 58:1230-40. 2014
    ..We set out to establish a new human challenge model and ascertain the S. Typhi (Quailes strain) inoculum required for an attack rate of 60%-75% in typhoid-naive volunteers when ingested with sodium bicarbonate solution...

Research Grants31

  1. Pacific NorthWest Regional Center of Excellence (PNWRCE)
    Jay A Nelson; Fiscal Year: 2013
    ..pseudomallei host pathogen response during both the septicemic as well as the intracellular phases of the disease. ..
  2. Southeast Regional Centers of Excellence for Biodefense &Emerging Infectious Di
    Philip Frederick Sparling; Fiscal Year: 2013
    ..SERCEB brings new investigators to the biodefense effort through a combination of educational programs, support of innovative new projects, and the synergistic interactions among its world-class investigators. ..
  3. New England Regional Center of Excellence in Biodefense and Emerging Infectious D
    Dennis L Kasper; Fiscal Year: 2013
    ..NERCE will also continue its Developmental Projects program and Career Development in Biodefense program in an effort to initiate new research efforts and to attract new investigators to this field. ..
  4. Northeast Biodefense Center
    W Ian Lipkin; Fiscal Year: 2013
    ..As a Center based in a School of Public Health and a State Department of Health, the NBC has a firm commitment to and practical understanding of Emergency Preparedness. ..
  5. Molecular Analyses and Interventions for Biodefense and Emerging Pathogens
    Olaf Schneewind; Fiscal Year: 2013
    ..Research and training at the GLRCE is governed by a mechanism involving ongoing review of scientific excellence and translational goals, inter-institutional advisory boards and external scientific advisory bodies. ..
  6. Pacific Southwest RCE for Biodefense &Emerging Infectious Diseases Research
    Alan G Barbour; Fiscal Year: 2013
    ..abstract_text> ..
  7. Rocky Mountain Regional Center of Excellence or Biodefense and Emerging Infectiou
    John T Belisle; Fiscal Year: 2013
    ..abstract_text> ..
  8. VACCINE INDUCED IMMUNITY IN THE YOUNG AND AGED
    Rafi Ahmed; Fiscal Year: 2013
    ....