Abstract

We identified a number of cell lines which can be transfected with RNA transcribed in vitro from our infectious cDNA clone and showed that the genomes replicate and that infectious virus is produced. HEV replicons expressing green fluorescent protein or luciferase were used to compare the effects of mutations in a non-coding region on replication in cell culture and in rhesus macaques. We showed that a viral protein that is phosphorylated in vitro is not required for genomic replication in cell culture but is required for infection of macaques, although it need not be phosphorylated. We developed a cell culture system which permits limited infection by wild-type HEV and used it to study neutralization and to show that HEV is more heat labile than is hepatitis A virus. We showed that a cDNA clone of swine HEV developed by a former post-doctoral student can replicate in our cell culture systems. We studied the humoral immune response to the HEV recombinant vaccine we developed previously and mapped the neutralization site, demonstrated that the vaccine induces antibodies that cross-react with viruses from all four genotypes, and showed that the vaccine contains all the immunodominant epitopes located in the capsid protein.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Intramural Research (Z01)
Project #
1Z01AI000596-14
Application #
6985878
Study Section
(LID)
Project Start
Project End
Budget Start
Budget End
Support Year
14
Fiscal Year
2004
Total Cost
Indirect Cost

  Institution

Name
Niaid Extramural Activities
Department
Type
DUNS #
City
State
Country
United States
Zip Code

  Publications

Graff, Judith; Zhou, Yi-Hua; Torian, Udana et al. (2008) Mutations within potential glycosylation sites in the capsid protein of hepatitis E virus prevent the formation of infectious virus particles. J Virol 82:1185-94
Purcell, R H; Emerson, S U (2008) Hepatitis E: an emerging awareness of an old disease. J Hepatol 48:494-503
Yu, Claro; Zimmerman, Carl; Stone, Roger et al. (2007) Using improved technology for filter paper-based blood collection to survey wild Sika deer for antibodies to hepatitis E virus. J Virol Methods 142:143-50
Zhou, Yi-Hua; Chen, Zhaochun; Purcell, Robert H et al. (2007) Positive reactions on Western blots do not necessarily indicate the epitopes on antigens are continuous. Immunol Cell Biol 85:73-8
Chen, Zhaochun; Earl, Patricia; Americo, Jeffrey et al. (2006) Chimpanzee/human mAbs to vaccinia virus B5 protein neutralize vaccinia and smallpox viruses and protect mice against vaccinia virus. Proc Natl Acad Sci U S A 103:1882-7
Chen, Zhaochun; Moayeri, Mahtab; Zhou, Yi-Hua et al. (2006) Efficient neutralization of anthrax toxin by chimpanzee monoclonal antibodies against protective antigen. J Infect Dis 193:625-33
Graff, Judith; Torian, Udana; Nguyen, Hanh et al. (2006) A bicistronic subgenomic mRNA encodes both the ORF2 and ORF3 proteins of hepatitis E virus. J Virol 80:5919-26
Meky, Fatma A; Stoszek, Sonia K; Abdel-Hamid, Mohamed et al. (2006) Active surveillance for acute viral hepatitis in rural villages in the Nile Delta. Clin Infect Dis 42:628-33
Emerson, Suzanne U; Clemente-Casares, Pilar; Moiduddin, Nasser et al. (2006) Putative neutralization epitopes and broad cross-genotype neutralization of Hepatitis E virus confirmed by a quantitative cell-culture assay. J Gen Virol 87:697-704
Stoszek, Sonia K; Abdel-Hamid, Mohamed; Saleh, Doa'a A et al. (2006) High prevalence of hepatitis E antibodies in pregnant Egyptian women. Trans R Soc Trop Med Hyg 100:95-101

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