Open AccessTranslocation of a hydrocarbon fluorescent probe between Epstein-Barr virus and lymphoid cells: An assay for early events in viral infection
Author(s) -
Ken S. Rosenthal,
Saul Yanovich,
Michael Inbar,
Jack L. Strominger
Publication year - 1978
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.75.10.5076
Subject(s) - chromosomal translocation , microbiology and biotechnology , liposome , diphenylhexatriene , virus , chemistry , epstein–barr virus , incubation , phosphatidylcholine , raji cell , biology , virology , fluorescence anisotropy , biochemistry , membrane , phospholipid , gene
Translocation of the hydrocarbon fluorescent probe diphenylhexatriene (DPH) between membranes was studied by fluorescence polarization (P ) analysis. First, using a model system, the highP value (0.324) of DPH-labeled cholesterol/phosphatidylcholine liposomes and the lowP value (0.157) of DPH-labeled phosphatidylcholine liposomes allowed detection of DPH translocation between interacting liposomes. This was monitored by the change inP in either direction. Early events during cell-virus interactions were similarly studied by monitoring DPH translocation. TheP value of DPH-labeled Epstein-Barr Virus (EBV) was significantly higher (0.350-0.392) than theP value of DPH-labeled lymphoid cells (0.238-0.289). Hence, DPH translocation could be detected by changes inP following incubation of DPH-labeled EBV and nonlabeled cells. A marked decrease inP was observed after incubation of DPH-labeled EBV with either nonlabeled lymphoblastoid Raji cells or fresh human B lymphocytes. However, only a slight decrease inP was obtained when DPH-labeled EBV was incubated with either nonlabeled fresh human T lymphocytes or fresh T or B rabbit lymphocytes. Moreover, incubation of fresh human B lymphocytes with the purified C3 component of complement (a putative inhibitor for the EBV receptor) prior to the addition of DPH-labeled EBV abolished the observed decrease in theP value. Most of these experiments were carried out with both the P3HR-1 and the B95-8 strains of EBV. DPH translocation, as determined by fluorescence polarization analysis, is, therefore, measuring some early event during interaction of this enveloped virus and mammalian cells. The potential applicability of this technique to other viruses is illustrated by an experiment with Semliki Forest virus.