Author
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Tian, Peng |
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JIANG, XI - UNIV. OF CINCINATI, OHIO |
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ZHONG, WEIMING - UNIV. OF CINCINATI, OHIO |
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JENSEN, HANNE - UNIV. OF CA, DAVIS |
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Brandl, Maria |
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Bates, Anne |
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Engelbrektson, Anna |
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Mandrell, Robert |
Submitted to: Research in Veterinary Science
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 1/30/2007 Publication Date: 12/1/2007 Citation: Tian, P., Jiang, X., Zhong, W., Jensen, H., Brandl, M., Bates, A.H., Engelbrektson, A.L., Mandrell, R.E. 2007. Binding of recombinant norovirus like particle to histo-blood group antigen on cells in the lumen of pig duodenum. Research in Veterinary Science. 83(3) 410-418 Interpretive Summary: Histo-blood group antigens (HBGA) expressed on cells in the human GI tract have been shown to function as receptors for noroviruses (NV). Previously, we have reported that mucin purified from pig stomach (PGM) could bind recombinant NV virus-like particles (rNVLP). We propose that similar HBGA presents in PGM and responsible for rNVLP binding. To address this question, specific anti-HBGA monoclonal antibodies (MAbs) were used to determine the presence of corresponding HBGA in PGM or pig gastro-intestine tissue (PGIT) washings and on epithelial cells. Type A, type H1, and Lewis b HBGAs in PGM were detected. Individual pigs were either HBGA type A positive or type H1 (type O) positive. rNVLPs from a genogroup I (rNW) or three genogroup II (rMOH, rVA207, and rVA387) strains could bind to plates coated with PGM or PGIT washings with similar binding patterns in humans. Recombinant Norwalk virus-like particles (rNW) were used for further characterization. rNW binding was inhibited by pre-incubating the wells with MAbs specific for either type A or type H1 HBGA, and by the presence of free HBGAs from human saliva. Co-localization of rNW and corresponding HBGA on PGIT epithelial cells was observed. These findings suggest that rNW binds to HBGAs expressed on PGIT epithelial cells. This is the first report of specific binding of human rNW to HBGAs in the PGIT. It highlights the importance of further study of human NV incidence and potential infection and residence in non-human animal hosts and suggests the possibility that NV may be a zoonotic pathogen. Technical Abstract: A set of HBGA specific monoclonal antibodies (MAbs) was used to determine the presence of corresponding HBGA in pig gastro-intestine tissue (PGIT) washings and on epithelial cells. . rNWs were applied to plate coated with pig gastro-intestine tissue (PGIT) washings , and measured by ELISA assay using polyclonal antibodies against NV. Co-localization of HBGA and rNW binding on epithelial cells of pig gastro-intestine tissue were determined by immuno-staining and analyzed in three-channel confocal microscopy. Type A, type H1, and Lewis b HBGAs in PGM were detected. Individual pigs were either HBGA type A positive or type H1 (type O) positive. rNVLPs from a genogroup I (rNW) or three genogroup II (rMOH, rVA207, and rVA387) strains could bind to plates coated with PGM or PGIT washings with similar binding patterns in humans. Recombinant Norwalk virus-like particles (rNW) were used for further characterization. rNW binding was inhibited by pre-incubating the wells with MAbs specific for either type A or type H1 HBGA, and by the presence of free HBGAs from human saliva. Co-localization of rNW and corresponding HBGA on PGIT epithelial cells was observed. These findings suggest that rNW binds to HBGAs expressed on PGIT epithelial cells. This is the first report of specific binding of human rNW to HBGAs in the PGIT. |