Intracellular Cleavage of Human Influenza A Virus Hemagglutinin and Its Inhibition |
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Authors: | O P Zhirnov I V Vorobjeva A V Ovcharenko H D Klenk |
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Institution: | (1) Russian Academy of Medical Sciences, ul. Gamalei 16, Ivanovsky Institute of Virology, Moscow, 123098, Russia;(2) Institute of Virology, Philipps University, Marburg, 35037, Germany |
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Abstract: | Replication of human influenza A viruses and proteolytic cleavage of the viral glycoprotein HA0 HA1/2 were studied in passaged cultures of epithelial cells of the serous membrane of human large intestine (CACO-2 line), dog kidney cells (MDCK), and monkey kidney cells (CV-1). Cleavage of the viral glycoprotein HA0, synthesis of activated virions, multicycle virus infection, and effective production of viral foci under an agarose overlayer were found in CACO-2 cells. By pulse–chase labeling of viral glycoproteins, testing the sensitivity to endoglycosidase-H of the viral glycoproteins HA0 and HA1/2 synthesized, and inhibiting the HA0 proteolysis with brefeldin A, the HA0 HA1/2 proteolysis was established to occur in the late stages of intracellular transport in the trans-Golgi and plasma membrane areas of the cells. Proteolysis of the viral glycoprotein HA0 in CACO-2 cells was suppressed by aprotinin, a natural inhibitor of serine proteinases. Unlike MDCK and CV-1 cells resistant to apoptosis induced by influenza virus, CACO-2 cells retained their viability for 2-3 days after infection with human influenza A virus. |
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Keywords: | influenza virus replication hemagglutinin cleavage aprotinin |
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