Effect of Certain Inhibitors of Glycoprotein Synthesis on Cell Fusion Induced by Vesicular Stomatitis Virus

Abstract

The effect of certain metabolic inhibitors on the fusion of BHK-21 cells induced by vesicular stomatitis virus (VSV) was studied. The polykaryocyte formation in infected cells and virus growth were inhibited by 2-deoxy-D-glucose and Vesicular stomatitis virus (VSV) grows rapidly and is highly cytopathic to several lines of mammalian cell culture (17). Recently it has been reported that marked polykaryocyte formation was observed in monolayers of some cell lines infected with VSV and that the virus replication caused cell fusion (3, 10, 15). These virus-cell interactions provided an excellent means for a detailed understanding of the fusion process and cytopathogenicity by using various anti-metabolites (13). The experiments were undertaken to analyse the fusion process that occurred during virus multiplication. MATERIALS AND METHODSCell cultures. Two strains of baby hamster kidney (BHK-21-KB and BHK-21-B) cells were grown in Eagle's minimum essential medium (MEM) containing 0.03% L-glutamine and 5% inactivated calf serum. The medium without glucose was used for studying the effect of 2-deoxy-D-glucose. The BHK-21-KB cells were most sensitive to VSV-induced cell fusion and BHK-21-B cells were less sensitive (15).

Shuttle of lentiviral vectors via transplanted cells in vivo

“…VSV-induced membrane fusion occurs at a very narrow pH range and can be avoided by different pH or other factors. [18][19][20] It is conceivable that the culture medium used during the transwell experiments allows release of already cellbound but not fused lentiviral particles. Whether this is a unique property of VSV-G pseudotyped viral particles or can also be observed when using other viral protein envelopes remains to be determined.…”

Requirement of Carbohydrates for Cell Fusion Induced by Vesicular Stomatitis Virus

“…On the basis of the results obtained previously (14,15), cell fusion induced by VSV in BHK-21-KB cells seemed to be related to specific carbohydrate metabolism. During studies on the inhibitory effect on cell fusion of 2-deoxy-n-glucose and n-glucosamine (14), it was found that polykaryocyte formation in VSV-infected BHK-21-KB cells was not induced in MEM without glucose, and the cytopathic effect was detected only by cell rounding or agglutination. From the results presented here, it can be seen that the rate of polykaryocyte formation was markedly reduced in a medium with less than 5.0 mM glucose while there was no reduction in virus production (Figs.…”

“…The virus titers were determined by plaque assay in BHK-21-KB cells. One to ten plaque-forming units (PFU) of virus per cell were inoculated and virus adsorption was allowed to proceed for 1 hr at 37 C (14) .…”

“…Our previous studies (14,IS) suggested that the cell fusion induced by VSV in BHK-21-KB cells was related to specific glycoprotein synthesis or carbohydrate metabolism. The present study shows that polykaryocyte formation in VSV-infected BHK-21 cells is not induced in Eagle's minimal essential medium (MEM) in the absence of glucose.…”

“…However, there is much less information regarding the details of biochemical analysis of the mechanism of cell fusion. Viral glycoproteins may play an important role in the process of cell fusion (7,12,14,IS). It has been shown that some sugar analogues interfere with the growth of a number of enveloped viruses (5,6), and they are also highly effective in preventing cell fusion by some viruses (3).…”