Abortive replication of influenza virus A/WSN/33 in HeLa229 cells: defective viral entry and budding processes.
Since influenza A virus replication is defective in HeLa229 cells but productive in Madin-Darby canine kidney (MDCK) cells, we have investigated the steps in the infectious cycle of A/WSN/33 virus defective in HeLa229 cells. We find that both the entry and exit processes of the infectious cycle were defective in HeLa229 cells. During entry, viral adsorption was apparently normal in HeLa229 cells but a subsequent step(s) involving one or more processes namely the fusion/uncoating and nuclear transport of viral ribonucleoprotein was inefficient and slow compared to those in MDCK cells. Fewer HeLa229 cells were infected at the same multiplicities of infection, resistance to ammonium chloride developed much more slowly and degradation of the incoming virus proteins was delayed when compared to those in MDCK cells. Subsequent to the entry process, there was no significant difference in either the synthesis of viral proteins or the transport, maturation, and membrane insertion of viral glycoproteins although the glycosylation pattern of hemagglutinin was different and the peak protein synthesis was albeit delayed in HeLa229 cells compared to that in MDCK cells. However, there was a major defect in the budding and release of viral particles. In HeLa229 cells, viral bud formation occurred but viral particles remained attached to the plasma membrane and were not released into the medium. This defect in virus release was not due to lack of neuraminidase activity but could be, at least partly, overcome by cytochalasin B treatment, suggesting a possible involvement of microfilaments in virus release. These results indicate that the abortive replication of influenza virus A/WSN/33 in HeLa229 cells appears to be due to multiple defects involving both the entry and release of viral particles and that host cell membrane and microfilaments may be important contributing factors in these processes.
Gujuluva
C N
CN
Department of Microbiology and Immunology, Jonsson Comprehensive Cancer Center, University of California at Los Angeles, School of Medicine 90024-1747.
Kundu
A
A
Murti
K G
KG
Nayak
D P
DP
D000818:Animals; D002478:Cells, Cultured; D003571:Cytochalasin B / Q000494:pharmacology; D003673:Defective Viruses / Q000502:physiology*; D004285:Dogs; D006367:HeLa Cells; D006801:Humans; D009980:Influenza A virus / Q000502:physiology*; D014764:Viral Proteins / Q000096:biosynthesis; D014771:Virion / Q000502:physiology; D014779:Virus Replication
D014764:Viral Proteins; D003571:Cytochalasin B