Present Homologous and Heterologous Antigen with Hepatitis E Virus

Summary

Principal Investigator: R Holland Cheng
Abstract: DESCRIPTION (provided by applicant): Advantages of DNA vaccines including well-tolerance, safety and ability to induce antibody, cytotoxic T- lymphocytes, and T-helper immune responses make it more and more attractive in treatment of chronic virus infections. In order to successfully induce immune response, DNA vaccine needs to enter target cell for transcription and subsequent protein production. An ideal delivery carrier needs to be safe and effective in targeting specific cell, provide sufficient protection of DNA plasmid during transportation, as wel as low- responses to any existing self-immunity. Deficient viral particles, such as adenoviruses or retroviruses, offer an attractive option with great benefits in targeting and protection but majr drawbacks on strong self-immunity as well as the complexity in producing recombinant viral particles. Non-replicating virus-like particle derived from human hepatitis E virus (HEV-VLP) is empty icosahedral cage composed of 60 copies of recombinant HEV capsid proteins. This VLP is capable of encapsulating DNA vaccine in vitro, delivering DNA vaccine to epithelium cell at gastrointestinal tract, and inducing antigen-specific humoral and cellular immune responses. The simple procedure in DNA encapsulation makes HEV-VLP of particular interest as gene carrier;however, induction of self-immunity is still the hurdle in using HEV-VLP for therapeutical vaccination. Our studies on HEV-VLP crystal structure and antigenic structure reveal a structural modularity, with which HEV recombinant capsid proteins interplay between VLP assembly and antigen presentation. By carrying insertion of 15 amino acids at an antibody-binding site, the chimeric VLP reduces the reactivity to anti-HEV antibodies meanwhile retains the icosahedral assembly. This data suggests us a strategy to lower the reactivity of VLP to antibody- induced neutralization by structural alteration at antibody-binding sites, a mechanism that viruses have evolved to mediate their escape from host immune surveillance. The proposed experiments in this application include 1) using cryo-electron microscopy and image reconstruction to identify HEV-VLP surface flexible loops that critical to antibody interactions;2 inserting short peptide into the identified surface loops to create chimeric VLP with attenuate reactivity to HEV-VLP;3) both wild type and chimeric VLPs will be evaluated in delivery of DNA vaccine encoding the surface antigen of hepatitis B virus, as potential gene carrier for treatment of chronic progressive hepatitis B. Because HEV is an enteric transmitted virus, HEV-VLP is able to transcytose the mucosal barrier at gastrointestinal tract and target specific mucosal region for antigen production. When a short peptide bearing mucosal adhesion ligand is inserted into surface antigenic loops, the chimeric VLP is hypothesized to have strong specificity in targeting mucosal region, leading to potent induction of mucosal immunity. In collaboration with Dr Kit Lam and Dr Christopher Walker, two experts respectively in drug discovery and HBV vaccine development, we expect to obtain sufficient data leading us to repeated use of HEV-VLPs as carrier for clinical gene therapy.
Funding Period: -------------------- - --------------------
more information: NIH RePORT

Top Publications

  1. pmc Structure of hepatitis E virion-sized particle reveals an RNA-dependent viral assembly pathway
    Li Xing
    Department of Molecular and Cellular Biology, University of California, Davis, California 95616, USA
    J Biol Chem 285:33175-83. 2010
  2. ncbi Chimeric hepatitis E virus-like particle as a carrier for oral-delivery
    Pitchanee Jariyapong
    Department of Molecular and Cellular Biology, University of California, Davis, CA 95616, United States
    Vaccine 31:417-24. 2013
  3. pmc "OA02" peptide facilitates the precise targeting of paclitaxel-loaded micellar nanoparticles to ovarian cancer in vivo
    Kai Xiao
    Department of Biochemistry and Molecular Medicine, University of California Davis, Sacramento, California 95817, USA
    Cancer Res 72:2100-10. 2012
  4. pmc Calpains promote α2β1 integrin turnover in nonrecycling integrin pathway
    Nina Rintanen
    Department of Biological and Environmental Science Nanoscience Center, University of Jyvaskyla, FI 40351 Jyväskylä, Finland
    Mol Biol Cell 23:448-63. 2012
  5. pmc Quaternary structures of HIV Env immunogen exhibit conformational vicissitudes and interface diminution elicited by ligand binding
    Carlos G Moscoso
    Department of Molecular and Cellular Biology, University of California, Davis, CA 95616, USA
    Proc Natl Acad Sci U S A 108:6091-6. 2011
  6. pmc The structure of avian polyomavirus reveals variably sized capsids, non-conserved inter-capsomere interactions, and a possible location of the minor capsid protein VP4
    Peter S Shen
    Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602, USA
    Virology 411:142-52. 2011
  7. pmc Spatial configuration of hepatitis E virus antigenic domain
    Li Xing
    Molecular and Cellular Biology, University of California, 1 Shields Ave, Davis, CA 95616, USA
    J Virol 85:1117-24. 2011
  8. pmc Copper-doxorubicin as a nanoparticle cargo retains efficacy with minimal toxicity
    Azadeh Kheirolomoom
    Department of Biomedical Engineering, 451 East Health Sciences Drive, School of Medicine, University of California, Davis, Davis, California 95616, United States
    Mol Pharm 7:1948-58. 2010
  9. pmc Structural evidence of glycoprotein assembly in cellular membrane compartments prior to Alphavirus budding
    Pan Soonsawad
    Department of Molecular and Cellular Biology, University of California, Davis, CA 95616 8536, USA
    J Virol 84:11145-51. 2010
  10. pmc Structure of the arginine methyltransferase PRMT5-MEP50 reveals a mechanism for substrate specificity
    Meng Chiao Ho
    Department of Biochemistry, Albert Einstein College of Medicine of Yeshiva University, Bronx, New York, United States of America
    PLoS ONE 8:e57008. 2013

Detail Information

Publications10

  1. pmc Structure of hepatitis E virion-sized particle reveals an RNA-dependent viral assembly pathway
    Li Xing
    Department of Molecular and Cellular Biology, University of California, Davis, California 95616, USA
    J Biol Chem 285:33175-83. 2010
    ..The in vitro assembly further demonstrated that HEV capsid protein had the intrinsic ability to form decameric intermediate. Our data suggest that RNA binding is the extrinsic factor essential for the assembly of HEV native capsids...
  2. ncbi Chimeric hepatitis E virus-like particle as a carrier for oral-delivery
    Pitchanee Jariyapong
    Department of Molecular and Cellular Biology, University of California, Davis, CA 95616, United States
    Vaccine 31:417-24. 2013
    ..Therefore, the insertion of peptides at the surface antigenic site could allow VLPs to escape pre-existing anti-HEV humoral immunity...
  3. pmc "OA02" peptide facilitates the precise targeting of paclitaxel-loaded micellar nanoparticles to ovarian cancer in vivo
    Kai Xiao
    Department of Biochemistry and Molecular Medicine, University of California Davis, Sacramento, California 95817, USA
    Cancer Res 72:2100-10. 2012
    ..Therefore, OA02-targeted telodendrimers loaded with paclitaxel have great potential as a new therapeutic approach for patients with ovarian cancer...
  4. pmc Calpains promote α2β1 integrin turnover in nonrecycling integrin pathway
    Nina Rintanen
    Department of Biological and Environmental Science Nanoscience Center, University of Jyvaskyla, FI 40351 Jyväskylä, Finland
    Mol Biol Cell 23:448-63. 2012
    ..In conclusion, we identified a novel virus- and clustering-specific pathway that diverts α2β1 integrin from its normal endo/exocytic traffic to a nonrecycling, calpain-dependent degradative endosomal route...
  5. pmc Quaternary structures of HIV Env immunogen exhibit conformational vicissitudes and interface diminution elicited by ligand binding
    Carlos G Moscoso
    Department of Molecular and Cellular Biology, University of California, Davis, CA 95616, USA
    Proc Natl Acad Sci U S A 108:6091-6. 2011
    ..Structural features gleaned from our studies should aid the design of Env-based immunogens for inducement of potent broadly neutralizing antibodies against exposed conformational epitopes...
  6. pmc The structure of avian polyomavirus reveals variably sized capsids, non-conserved inter-capsomere interactions, and a possible location of the minor capsid protein VP4
    Peter S Shen
    Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602, USA
    Virology 411:142-52. 2011
    ..However, the plug density is more prominent in APV and may include VP4, a minor capsid protein unique to bird polyomaviruses...
  7. pmc Spatial configuration of hepatitis E virus antigenic domain
    Li Xing
    Molecular and Cellular Biology, University of California, 1 Shields Ave, Davis, CA 95616, USA
    J Virol 85:1117-24. 2011
    ..Therefore, the T=1 HEV VLP is a novel delivery system for displaying foreign epitopes at the VLP surface in order to induce antibodies against both HEV and the inserted epitope...
  8. pmc Copper-doxorubicin as a nanoparticle cargo retains efficacy with minimal toxicity
    Azadeh Kheirolomoom
    Department of Biomedical Engineering, 451 East Health Sciences Drive, School of Medicine, University of California, Davis, Davis, California 95616, United States
    Mol Pharm 7:1948-58. 2010
    ..After the 28-day course of therapy, syngeneic tumors regressed to a premalignant phenotype of ∼(1 mm)(3) or could not be detected...
  9. pmc Structural evidence of glycoprotein assembly in cellular membrane compartments prior to Alphavirus budding
    Pan Soonsawad
    Department of Molecular and Cellular Biology, University of California, Davis, CA 95616 8536, USA
    J Virol 84:11145-51. 2010
    ..Thus, the structural characteristics of CPV-II can be used in evaluating the design of a packaging cell line for replicon production...
  10. pmc Structure of the arginine methyltransferase PRMT5-MEP50 reveals a mechanism for substrate specificity
    Meng Chiao Ho
    Department of Biochemistry, Albert Einstein College of Medicine of Yeshiva University, Bronx, New York, United States of America
    PLoS ONE 8:e57008. 2013
    ..Electron microscopy and reconstruction showed substrate centered on MEP50. These data support a mechanism in which MEP50 binds substrate and stimulates PRMT5 activity modulated by substrate post-translational modifications...