Core B, as part of the Program Project: SARS Coronavirus: Inhibition of Entry with Dr. K. V. Holmes as PI,will provide BSL3 support when required primarily for Projects #1, #2 and #6. The plan is to evolve Core Bfrom a laboratory dedicated to SARS work within the Bio-Environmental Hazards Research Building(BHRB) at the Foothills Campus of Colorado State University (CSU) in conjunction with a contract recentlyfunded by NIH for investigation of SARS as a new unit within a new research program on emerging viraldiseases administered by Dr. Barry Beaty at CSU. The new SARS laboratory is expected to be functioningbefore review of this proposal is completed.
Specific Aims for Core B: (1) Propagation, titration andpurification of SARS-CoV. (2) Assay antibody-mediated detection of virus infection in cells by viralcytopathic effects, immunofluoresence, immunocytochemistry or FACS bioanalyzer. (3) Assay antibody-mediated inhibition of binding or infection of live virus with permissive host cells. (4) Assay viral S peptide-mediated inhibition of binding or infection of live virus with permissive host cells. (5) Prepare tissues (e.g.brush border membranes) from a variety of species for binding studies with SARS-CoV. (6) Extract SARS-CoV RNA to aid cloning of receptor cDNA. (7) Test anti-viral drugs or vaccines that target SARS-CoV-receptor interactions in animals. Core B will have an integral role in the Program Project achieving the goalsof finding several complementary ways to block virus binding to susceptible cells and thereby prevent virusinfection. Improved diagnosis, treatment and control of SARS-CoV infection in humans are dependent uponlearning which cells or tissues bind the virus. Information derived from infection of naturally occurringsusceptible animal models, or transgenic mice with genetically induced susceptibility will be used to testexperimental treatment protocols that interfere with virus binding to receptors. The findings from theProgram Project are expected to provide guidance for treatment of SARS-infected humans.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Program Projects (P01)
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Special Emphasis Panel (ZAI1)
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University of Colorado Denver
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Zhou, Bin; Pearce, Melissa B; Li, Yan et al. (2013) Asparagine substitution at PB2 residue 701 enhances the replication, pathogenicity, and transmission of the 2009 pandemic H1N1 influenza A virus. PLoS One 8:e67616
Zhou, Bin; Wentworth, David E (2012) Influenza A virus molecular virology techniques. Methods Mol Biol 865:175-92
Funk, C Joel; Wang, Jieru; Ito, Yoko et al. (2012) Infection of human alveolar macrophages by human coronavirus strain 229E. J Gen Virol 93:494-503
Chen, Lanfen; Chen, Zhangguo; Baker, Kristi et al. (2012) The short isoform of the CEACAM1 receptor in intestinal T cells regulates mucosal immunity and homeostasis via Tfh cell induction. Immunity 37:930-46
Zhou, Bin; Li, Yan; Speer, Scott D et al. (2012) Engineering temperature sensitive live attenuated influenza vaccines from emerging viruses. Vaccine 30:3691-702
Zhou, Bin; Jerzak, Greta; Scholes, Derek T et al. (2011) Reverse genetics plasmid for cloning unstable influenza A virus gene segments. J Virol Methods 173:378-83
Zhou, Bin; Li, Yan; Halpin, Rebecca et al. (2011) PB2 residue 158 is a pathogenic determinant of pandemic H1N1 and H5 influenza a viruses in mice. J Virol 85:357-65
Peng, Guiqing; Sun, Dawei; Rajashankar, Kanagalaghatta R et al. (2011) Crystal structure of mouse coronavirus receptor-binding domain complexed with its murine receptor. Proc Natl Acad Sci U S A 108:10696-701
Osborne, Christina; Cryan, Paul M; O'Shea, Thomas J et al. (2011) Alphacoronaviruses in New World bats: prevalence, persistence, phylogeny, and potential for interaction with humans. PLoS One 6:e19156
Zhou, Bin; Li, Yan; Belser, Jessica A et al. (2010) NS-based live attenuated H1N1 pandemic vaccines protect mice and ferrets. Vaccine 28:8015-25

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