Abstract
This chapter describes the binding, endocytosis, and degradation of enveloped animal viruses. The assays described in this chapter are designed to monitor the interaction of viruses with monolayer tissue culture cells. The techniques used in studies with Semliki Forest virus (SFV) and baby hamster kidney (BHK21) cells are described in addition to the adapted procedures for vesicular stomatitis virus (VSV) and fowl plague virus (FPV) interaction with Madin–Darby canine kidney (MDCK) cells. To study virus binding on live cells, low temperatures must be used to prevent endocytosis. Two types of binding medium (BM), RPM 1-1640 and G-MEM, are used. Virus binding to cells is highly pH sensitive and careful buffering is essential: the absence of bicarbonate enables use of the media in normal atmosphere without changes in pH. Many enveloped animal viruses have potent low pH-induced membrane fusion activities. The nucleocapsids of SFV are ribonuclease (RNase) sensitive. This property can be exploited to measure the penetration of the viral nucleocapsids to the cytosol and the release of the nucleocapsids from the protective viral envelope.
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