Analysis of total lipid, phospholipid and cholesterol distribution has been conducted in parallel on BHK 21/13S cells grown in suspension cultures, on purified Rubella virus and on cells infected by the virus. Extracellular virus was purified by use of a previously described procedure (3,4). A higher content of lipid and phospholipid was found in infected cells (versus control cells) which were characterized by the presence of an unidentified nonphosphorylated lipid fraction that was detected neither in the control cells nor in the purified virus. The level and the nature of phospholipids and cholesterol of BHK 21/13S cells (infected or not) were compared to those of various clones of BHK 21cells. The same phospholipids were detected in the virus and in the cells but phosphatidyl choline level was much higher than in the control cells and lower than in the infected cells, while phosphatidyl ethanolamine content was lower than in the cells (infected or not). The presence of cardiolipin (4.4 per cent), the amount of sphingomyelin (6.9 per cent) and the molar ratio of cholesterol to phospholipids (0.26) in varions seem to favor a rubella virus maturation site in the cells.
When inoculated at the same MOI, Vero cells released a larger amount of infectious rubella virus into the culture medium than did BHK21 cells. However, BHK21 cells (in monolayer or in suspension) produced more intracellular infectious virus than Vero cells when tested 24 h after infection. Maturation of the virus in BHK21 cells occurred at the plasma membrane and, in a larger quantity, in the cytoplasm (Golgi apparatus and vacuoles). Viral particles consisted of an electron-dense core (32 nm) surrounded by a capsid and enveloped by a single membrane (8–10 nm). Aberrant forms (elongated and twisted) in the vacuole and double virions in the plasma membrane were observed as early as 65 h after infection.
A simple and reproducible method for the production of purified rubella virus is described. Purified virus was subjected to morphological and chemical analysis. The virus particles were rather pleomorphic (60 nm diameter), sometimes with one or more peripheral protrusions. The viral surface, revealed by negative staining, was composed of spikes 6 nm long, featuring enlarged ends. In SDS-urea-polyacrylamide gel electrophoresis, 4 major and 4 minor polypeptide bands were revealed. Total lipids and phospholipids were analysed on the same preparation. The viral particles were composed of RNA: 0.030 mg, and lipids: 0.245 mg, of which 0.169 mg were phospholipids for each mg of viral protein. Biologically, the purified virus preparation showed high infectivity, a high hemagglutination titre and a weak neuraminidase activity under defined conditions.
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