Viral Gastroenteritis: Basis of Protection and Virulence

Summary

Principal Investigator: HARRY BERNARD GREENBERG
Abstract: DESCRIPTION (provided by applicant): Rotavirus (RV) infection is the leading cause of childhood diarrheal morbidity and mortality worldwide despite the recent approval of two safe and effective live attenuated vaccines. Many fundamental questions concerning the basis of heterotypic humoral immunity, the mechanisms by which RV elicits but also avoids the innate immune response, and the factors controlling tissue tropism, systemic replication, and host range restriction remain to be answered. In this proposal, we will use a series of related and complementary approaches to characterize the mechanisms by which specific components of the virus and the host immune systems interact to either enhance or restrict RV replication and pathogenesis in the host. These aims are to: 1) Define the clonal and structural basis of heterotypic humoral rotavirus immunity in humans. Heterotypic (serotype cross-reactive) immunity plays an important role in preventing RV disease and is a critical contributor to the efficacy of at least one of the two currently licensed RV vaccines. Heterotypic humoral immunity is also a key component of several established (e.g., influenza) and experimental (e.g., HIV and HCV) vaccines. We propose to define, at the individual immunoglobulin (Ig) molecule level, the serologic nature of heterotypic neutralizing reactivity and the structural basis for this reactivity. Our hypothesis is that humans circumvent the serotypic diversity of circulating RV strains by developing lg molecules directed at VP4, VP7, and VP6 that mediate heterotypic protection. 2) Identify RV genes responsible for variation between strains in their ability to replicate systemically and cause systemic disease and characterize the link between interferon (IFN) interference and cellular tropism. RV spreads and replicates systemically although strains vary substantially in both the level and sequaelae of the systemic phase of infection. IFN signaling appears to play an important role in regulating systemic replication of different RV strains. We will determine, in vivo, which RV genes are responsible for the variation in systemic replication capacity and distinct organ specificity of different RV strains and investigate how the strain-dependent effects of NSP1 on IFN signaling affect this cell tropism and host range. Our hypothesis is that the NSP1 protein (gene 5 product), through its host-specific inhibitory effects on IFN signaling, plays an important, but not exclusive role in regulating cell tropism and systemic RV replication. 3) Characterize the interaction of murine plasmacytoid dendritic cells (pDCs) with RV and determine the contribution of pDC-derived interferon to restricting RV infection. Recent studies in humans and animals demonstrated that RV infection is not limited to the mature epithelial tip cells of the small bowel but also occurs in other cells at other sites, including DCs. pDCs are a primary source of the host anti- viral IFN response and studies by us and others provide support for the notion that IFN is an important regulator of RV pathogenesis that specifically functions to restrict systemic replication. Here we will characterize the interaction between murine pDCs and RV in vitro and use an in vivo mouse model to determine the role of pDCs in restricting systemic replication as well as B cell activation. Our hypothesis is that RV interaction with pDCs plays a critical role in restricting systemic RV replication and that pDCs, unlike many other host cells, are capable of circumventing RV's inherent ability to block the induction of IFN. PUBLIC HEALTH RELEVANCE: Rotaviruses are important in their own right since they kill more than half a million children per year. They also represent a highly tractable system to study basic aspects of innate, acquired and mucosal immunity and viral pathogenesis, all important issues for the future of infectious disease prevention and treatment.
Funding Period: 1984-07-01 - 2015-01-31
more information: NIH RePORT

Top Publications

  1. ncbi Immunity and correlates of protection for rotavirus vaccines
    Manuel A Franco
    Instituto de Genetica Humana, Pontificia Universidad Javeriana, Bogota, Colombia
    Vaccine 24:2718-31. 2006
  2. pmc The early interferon response to rotavirus is regulated by PKR and depends on MAVS/IPS-1, RIG-I, MDA-5, and IRF3
    Adrish Sen
    Department of Microbiology, Stanford University, Stanford, CA 94305 5101, USA
    J Virol 85:3717-32. 2011
  3. pmc Reconciliation of rotavirus temperature-sensitive mutant collections and assignment of reassortment groups D, J, and K to genome segments
    Jeanette Criglar
    Department of Molecular Virology and Microbiology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA
    J Virol 85:5048-60. 2011
  4. pmc Cross-linking of rotavirus outer capsid protein VP7 by antibodies or disulfides inhibits viral entry
    Scott T Aoki
    Laboratory of Molecular Medicine, Children s Hospital, 3 Blackfan Circle, Boston, MA 02115, USA
    J Virol 85:10509-17. 2011
  5. pmc Rhesus rotavirus trafficking during entry into MA104 cells is restricted to the early endosome compartment
    Marie Wolf
    Departments of Medicine and Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, USA
    J Virol 86:4009-13. 2012
  6. pmc Rotavirus immune responses and correlates of protection
    Juana Angel
    Instituto de Genetica Humana, Pontificia Universidad Javeriana, Bogota, Colombia
    Curr Opin Virol 2:419-25. 2012
  7. pmc Human rotavirus-specific IgM Memory B cells have differential cloning efficiencies and switch capacities and play a role in antiviral immunity in vivo
    Carlos F Narvaez
    Instituto de Genetica Humana, Facultad de Medicina, Pontificia Universidad Javeriana, Bogota, Colombia
    J Virol 86:10829-40. 2012
  8. pmc Innate immune response to homologous rotavirus infection in the small intestinal villous epithelium at single-cell resolution
    Adrish Sen
    Department of Microbiology, Stanford University, Stanford, CA 94305, USA
    Proc Natl Acad Sci U S A 109:20667-72. 2012
  9. pmc The battle between rotavirus and its host for control of the interferon signaling pathway
    Michelle M Arnold
    Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, United States of America
    PLoS Pathog 9:e1003064. 2013
  10. pmc Combinatorial tetramer staining and mass cytometry analysis facilitate T-cell epitope mapping and characterization
    Evan W Newell
    Agency for Science, Technology and Research, Singapore Immunology Network, Singapore
    Nat Biotechnol 31:623-9. 2013

Research Grants

Detail Information

Publications25

  1. ncbi Immunity and correlates of protection for rotavirus vaccines
    Manuel A Franco
    Instituto de Genetica Humana, Pontificia Universidad Javeriana, Bogota, Colombia
    Vaccine 24:2718-31. 2006
    ....
  2. pmc The early interferon response to rotavirus is regulated by PKR and depends on MAVS/IPS-1, RIG-I, MDA-5, and IRF3
    Adrish Sen
    Department of Microbiology, Stanford University, Stanford, CA 94305 5101, USA
    J Virol 85:3717-32. 2011
    ..These findings reveal that activation of the antiviral response by rotavirus is dependent on MAVS/IPS-1 and IRF3 and involves both RIG-I and MDA-5 and that IFN-β secretion during rotavirus infection is regulated by PKR...
  3. pmc Reconciliation of rotavirus temperature-sensitive mutant collections and assignment of reassortment groups D, J, and K to genome segments
    Jeanette Criglar
    Department of Molecular Virology and Microbiology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA
    J Virol 85:5048-60. 2011
    ..Rotavirus ts mutation groups are now mapped to 9 of the 11 rotavirus genome segments. Possible segment locations of the two remaining unmapped ts mutant groups are discussed...
  4. pmc Cross-linking of rotavirus outer capsid protein VP7 by antibodies or disulfides inhibits viral entry
    Scott T Aoki
    Laboratory of Molecular Medicine, Children s Hospital, 3 Blackfan Circle, Boston, MA 02115, USA
    J Virol 85:10509-17. 2011
    ..We conclude that dissociation of the VP7 trimer is an essential step in viral penetration into cells...
  5. pmc Rhesus rotavirus trafficking during entry into MA104 cells is restricted to the early endosome compartment
    Marie Wolf
    Departments of Medicine and Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, USA
    J Virol 86:4009-13. 2012
    ..These data suggest that early RRV trafficking is confined to the early endosome compartment and requires Rab5...
  6. pmc Rotavirus immune responses and correlates of protection
    Juana Angel
    Instituto de Genetica Humana, Pontificia Universidad Javeriana, Bogota, Colombia
    Curr Opin Virol 2:419-25. 2012
    ..Protection against rotavirus following vaccination is substantially heterotypic; nonetheless, a role for homotypic immunity in selection of circulating postvaccination strains needs further study...
  7. pmc Human rotavirus-specific IgM Memory B cells have differential cloning efficiencies and switch capacities and play a role in antiviral immunity in vivo
    Carlos F Narvaez
    Instituto de Genetica Humana, Facultad de Medicina, Pontificia Universidad Javeriana, Bogota, Colombia
    J Virol 86:10829-40. 2012
    ..Thus, RV-IgM(+) mBc are heterogeneous, occur more frequently than estimated by traditional limiting dilution analysis, have the capacity to switch Ig class in vitro as well as in vivo, and can mediate systemic antiviral immunity...
  8. pmc Innate immune response to homologous rotavirus infection in the small intestinal villous epithelium at single-cell resolution
    Adrish Sen
    Department of Microbiology, Stanford University, Stanford, CA 94305, USA
    Proc Natl Acad Sci U S A 109:20667-72. 2012
    ..Resolution of "averaged" innate immune responses in single IECs thus revealed unexpected heterogeneity in both the induction and subversion of early host antiviral immunity, which modulated host range...
  9. pmc The battle between rotavirus and its host for control of the interferon signaling pathway
    Michelle M Arnold
    Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, United States of America
    PLoS Pathog 9:e1003064. 2013
    ..In this review, we discuss the nature of the innate antiviral responses triggered by rotavirus infection and the viral mechanisms for inhibiting these responses...
  10. pmc Combinatorial tetramer staining and mass cytometry analysis facilitate T-cell epitope mapping and characterization
    Evan W Newell
    Agency for Science, Technology and Research, Singapore Immunology Network, Singapore
    Nat Biotechnol 31:623-9. 2013
    ..This approach should be useful for the comprehensive analysis of T-cell responses to infectious diseases or vaccines. ..
  11. pmc Characterization of rotavirus RNAs that activate innate immune signaling through the RIG-I-like receptors
    Dina Uzri
    Departments of Medicine and Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, USA
    PLoS ONE 8:e69825. 2013
    ....
  12. pmc Roles of VP4 and NSP1 in determining the distinctive replication capacities of simian rotavirus RRV and bovine rotavirus UK in the mouse biliary tract
    Ningguo Feng
    Stanford University, Stanford, California, USA
    J Virol 85:2686-94. 2011
    ..Our data indicate that systemic rotavirus strain-specific replication in the murine biliary tract is determined by both viral entry mediated by VP4 and viral antagonism of the host innate immune response mediated by NSP1...
  13. pmc Rhesus rotavirus entry into a polarized epithelium is endocytosis dependent and involves sequential VP4 conformational changes
    Marie Wolf
    Stanford University, VA PAHCS, 3801 Miranda Avenue 154 C, Building 101, Room C4 181, Palo Alto, CA 94304, USA
    J Virol 85:2492-503. 2011
    ..Together, these findings suggest that internalization, decapsidation, and cell membrane penetration involve endocytosis, calcium-dependent uncoating, and VP4 conformational changes, including a fold-back...
  14. pmc Rotavirus structural proteins and dsRNA are required for the human primary plasmacytoid dendritic cell IFNalpha response
    Emily M Deal
    Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, United States of America
    PLoS Pathog 6:e1000931. 2010
    ....
  15. pmc Dissecting rotavirus particle-raft interaction with small interfering RNAs: insights into rotavirus transit through the secretory pathway
    Mariela A Cuadras
    Division of Gastroenterology and Hepatology, Department of Medicine, Stanford University School of Medicine, California 94305, USA
    J Virol 80:3935-46. 2006
    ....
  16. pmc Extraintestinal spread and replication of a homologous EC rotavirus strain and a heterologous rhesus rotavirus in BALB/c mice
    M Fenaux
    Stanford University School of Medicine, 300 Pasteur Drive, Stanford, CA 94305, USA
    J Virol 80:5219-32. 2006
    ....
  17. pmc Role of interferon in homologous and heterologous rotavirus infection in the intestines and extraintestinal organs of suckling mice
    N Feng
    Stanford University, Stanford, California, USA
    J Virol 82:7578-90. 2008
    ....
  18. pmc The influence of CD4+ CD25+ Foxp3+ regulatory T cells on the immune response to rotavirus infection
    Bumseok Kim
    Department of Medicine and Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305, USA
    Vaccine 26:5601-11. 2008
    ..Our studies of immune modulatory Treg cells in the RV infection model may promote a better understanding of the basis for RV immunity as well as providing valuable clues for the development of more immunogenic RV vaccines...
  19. pmc Characterization of rotavirus specific B cells and their relation with serological memory
    Olga Lucía Rojas
    Instituto de Genetica Humana, Pontificia Universidad Javeriana, Carrera 7 40 62, Bogota, Colombia
    Virology 380:234-42. 2008
    ..Our studies contribute to understand the relationship between circulating mBc and serological memory, and enhance our capacity to develop better correlates of protection against RV disease...
  20. pmc Variation in antagonism of the interferon response to rotavirus NSP1 results in differential infectivity in mouse embryonic fibroblasts
    N Feng
    Stanford University, Stanford, California, USA
    J Virol 83:6987-94. 2009
    ..Therefore, there is a direct relationship between the replication efficiencies of different rotavirus strains in MEFs and strain-related variations in NSP1-mediated antagonism of the type I IFN response...
  21. pmc Rotaviruses: from pathogenesis to vaccination
    Harry B Greenberg
    Department of Medicine and Microbiology and Immunology, Stanford University School of Medicine, Stanford, California 94305 5119, USA
    Gastroenterology 136:1939-51. 2009
    ..Molecular biology studies of rotavirus replication and pathogenesis have identified unique viral targets that might be useful in developing therapies for immunocompromised children with chronic infections...
  22. pmc IRF3 inhibition by rotavirus NSP1 is host cell and virus strain dependent but independent of NSP1 proteasomal degradation
    Adrish Sen
    Department of Microbiology and Immunology, Stanford University, Stanford, CA 94305 5119, USA
    J Virol 83:10322-35. 2009
    ..Thus, NSP1's ability to degrade IRF3 is host cell dependent and is independent of NSP1 proteasomal degradation...
  23. pmc VP5* rearranges when rotavirus uncoats
    Joshua D Yoder
    Laboratory of Molecular Medicine, Children s Hospital, Boston, Massachusetts 02115, USA
    J Virol 83:11372-7. 2009
    ..Using one of these antibodies, we demonstrate that rotavirus uncoating triggers a conformational change in the cleaved VP4 spike to yield rearranged VP5*...
  24. pmc Rotavirus differentially infects and polyclonally stimulates human B cells depending on their differentiation state and tissue of origin
    Carlos F Narvaez
    Department of Medicine, Stanford University School of Medicine, 300 Pasteur Dr, Alway Bldg, Rm M121, Stanford, CA 94305, USA
    J Virol 84:4543-55. 2010
    ..Thus, RV differentially interacts with primary human B cells depending on their tissue of origin and differentiation stage, and it affects their capacity to modulate the local and systemic immune responses...
  25. pmc Rotavirus NSP1 protein inhibits interferon-mediated STAT1 activation
    Adrish Sen
    Department of Microbiology and Immunology, Stanford University, Stanford, California, USA
    J Virol 88:41-53. 2014
    ..This property segregated with the NSP1 gene and was observed in a simian SA11 monoreassortant that encoded porcine OSU NSP1 but not in wild-type SA11 or a reassortant encoding simian RRV NSP1. ..

Research Grants30

  1. Rotavirus: Studies of Intestinal Tropism, Viral Entry, and Rescue
    HARRY BERNARD GREENBERG; Fiscal Year: 2013
    ..It is our hypothesis that we can accomplish this aim by utilizing approaches recently validated for reovirus and orbiviruses. ..