Epstein-Barr virus infectivity of Raji and Molt 4 cells: differences in cellular membrane lipids and apparent microviscosity.
Infection of lymphocytes by the Epstein-Barr virus (EBV) is initiated by attachment of the major virus glycoprotein gp350/220 to a cell surface glycoprotein, known as CR2 (CD21). In a productive infection the virus envelope fuses with host cell membranes either at the cell surface or within endocytic vesicles. To investigate the relevance of host cell membrane properties in the fusion process, we used the lymphoblastoid cell lines Raji and Molt 4. Both cell lines express CR2 and bind EBV; however, only the Raji cell supports virus-cell fusion. Lipid analysis of the two cell lines indicated that Raji cells had a significantly lower cholesterol to phospholipid molar ratio due to a greater membrane content of phospholipid relative to protein. Determination of cell membrane fluid dynamics by fluorescence polarization indicated that the apparent membrane microviscosity of Molt 4 cells was significantly greater than that of Raji. Increasing Raji cell membrane apparent microviscosity to values similar to those of Molt 4 cells by incubation with cholesteryl-hemisuccinate caused a reduction in EBV fusion with Raji cells. However, experiments designed to allow EBV infection of Molt 4 cells whose plasma membranes had been fluidized were unsuccessful. These studies suggest that the lipid composition and other as yet unidentified factors are involved in entry of EBV into cells.
Department of Pharmacology and Experimental Therapeutics, College of Medicine, University of Florida, Gainesville 32610.
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Patel
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Hutt-Fletcher
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Crews
D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism / Q000382:microbiology*; D002788:Cholesterol Esters / Q000494:pharmacology; D004161:Diphenylhexatriene / Q000031:analogs & derivatives; D005456:Fluorescent Dyes; D004854:Herpesvirus 4, Human / Q000472:pathogenicity* / Q000502:physiology; D006801:Humans; D007700:Kinetics; D008560:Membrane Fluidity; D008561:Membrane Fusion; D008563:Membrane Lipids / Q000378:metabolism*; D014779:Virus Replication / Q000187:drug effects; D014783:Viscosity
D002788:Cholesterol Esters; D005456:Fluorescent Dyes; D008563:Membrane Lipids; D004161:Diphenylhexatriene; C033628:1-(4-(trimethylamino)phenyl)-6-phenylhexa-1,3,5-triene; C013440:cholesteryl succinate