Studies on the behavior of hantavirus envelope glycoprotein in adsorption and invasion to target cells.

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Hantaviruses are causative agents for rodent-borne viral zoonosis, HFRS and HPS. Envelope glycoproteins G1 and G2 induce neutralizing antibodies. Monoclonal antibodies possessing neutralizing activities showed fusion inhibition activities. This means vaccine target epitope for hantavirus is a fusion responsible region in envelope protein. I studied the role of envelope glycoproteins in establishment of infection in cells. At the first, we found highly sensitive cell lines (A549, BHK and VeroE6 cells) and non-permissive cell lines (MDBK) by using pseudotype VSV screening. AlphaVbeta3 integrin was reported as candidate of the cellular receptor for hantavirus entry. Therefore, we tried to induce alphaVbeta3 into MDBK. I could not detect change of infectivity in that cells. However, transfection efficiency of plasmid constract to MDBK cells were not enough to evaluate the effect of molecules. Otherwise, anti-beta3 integrin antibodies were not inhibit cellular entry of hantavirus in permissive cells found in this study. So that this molecules was considered not to be major receptor molecules in these cells. Next, I screened bindings of glycolipids from permissive cell line Vero E6 cells against hantavirus glycoprotein. Any significant binding was not detected between glycolipids and envelope proteins. These results suggested that the cellular receptor of hantavirus must be protein molecules except alphaVbeta3 integrin. Finally, a clone 5 of MDBK cells was established as relative high DNA induction and non-permissive line. And cDNA libraly from permissive cell line A5449 was produced. This study established the basis for analysis of interaction between hantavirus envelope glycoproteins and cellular molecules.