The mouse hepatitis virus A59 spike protein is not cleaved in primary hepatocyte and glial cell cultures.
Mouse hepatitis virus strain A59 (MHV-A59) produces mild meningoencephalitis and severe hepatitis during acute infection. To determine whether an in vitro system could be established which would mimic in vivo replication of the virus, we examined the ability of MHV-A59 to replicate in primary cultures of hepatocytes derived from C57BL/6 mice. Infection of hepatocytes with MHV-A59 resulted in low levels of replication, with virus remaining cell associated. Maximum viral yield was observed at 24 hours postinfection, while occasional syncytia were observed at 48 hours postinfection. Primary glial cell culture represents a potential in vitro system representing the second main target of MHV-A59, namely the brain. It is known that MHV-A59 produces a productive, but nonlytic infection in these cultures. Since cell-to-cell fusion is associated with the cleavage of S, the observation of little or no syncytia following MHV-A59 infection of both hepatocytes and glial cells prompted us to examine the cleavage of the spike protein (S) by Western blot analysis. The cleavage of S is inefficient in MHV-A59 infected hepatocytes and in glial cells. Furthermore, no cleavage of this protein was detected in liver homogenates from C57BL/6 mice infected with MHV-A59. These data suggest that cleavage of the MHV-A59 S protein, and by inference cell-to-cell fusion, does not seem to be essential for entry and spread of the virus in vivo and in vitro.
Department of Microbiology and Immunology, Philadelphia College of Osteopathic Medicine, Pennsylvania 19131, USA.
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Hingley
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Leparc-Goffart
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Weiss
D000818:Animals; D002478:Cells, Cultured; D007739:L Cells; D008099:Liver / Q000166:cytology; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D006517:Murine hepatitis virus / Q000378:metabolism* / Q000502:physiology; D009457:Neuroglia; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins / Q000378:metabolism*; D014779:Virus Replication
D008562:Membrane Glycoproteins; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins; C578557:spike glycoprotein, SARS-CoV; C578553:spike protein, mouse hepatitis virus