Intracellular cleavage of human influenza a virus hemagglutinin and its inhibition.
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 mucosal 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.
Zhirnov
O P
OP
Ivanovsky Institute of Virology, Russian Academy of Medical Sciences, Moscow 123098, Russia. zhirnov@virology.ru
Vorobjeva
I V
IV
Ovcharenko
A V
AV
Klenk
H D
HD
D000818:Animals; D017209:Apoptosis; D007611:Aprotinin / Q000494:pharmacology; D020126:Brefeldin A / Q000494:pharmacology; D002460:Cell Line; D004285:Dogs; D006023:Glycoproteins / Q000378:metabolism; D000882:Haplorhini; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000378:metabolism*; D006596:Hexosaminidases / Q000378:metabolism; D006801:Humans; D009980:Influenza A virus / Q000502:physiology*; D014779:Virus Replication
D006023:Glycoproteins; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D020126:Brefeldin A; D007611:Aprotinin; D006596:Hexosaminidases