Detection of receptor-specific murine leukemia virus binding to cells by immunofluorescence analysis.
Four classes of murine leukemia virus (MuLV) which display distinct cellular tropisms and bind to different retrovirus receptors to initiate virus infection have been described. In the present study, we describe a rapid, sensitive immunofluorescence assay useful for characterizing the initial binding of MuLV to cells. By using the rat monoclonal antibody 83A25 (L. H. Evans, R. P. Morrison, F. G. Malik, J. Portis, and W. J. Britt, J. Virol. 64:6176-6183, 1990), which recognizes an epitope of the envelope gp70 molecule common to the different classes of MuLV, it is possible to analyse the binding of ecotropic, amphotropic, or xenotropic MuLV by using only a single combination of primary and secondary antibodies. The MuLV binding detected by this assay is envelope receptor specific and matches the susceptibility to infection determined for cells from a variety of species. The binding of amphotropic MuLV to NIH 3T3 cells was shown to be rapid, saturable, and temperature dependent. Chinese hamster ovary (CHO-K1) cells normally lack the ability to bind ecotropic virus and are not infectible by ecotropic vectors. Expression of the cloned ecotropic retrovirus receptor gene (Rec) in CHO-K1 cells confers high levels of ecotropic virus-specific binding and confers susceptibility to infection. Characterization of MuLV binding to primary cells may provide insight into the infectibility of cells by retroviruses and aid in the selection of appropriate vectors for gene transfer experiments.
Kadan
M J
MJ
Gene Transfer Laboratory, Genetic Therapy, Inc., Gaithersburg, Maryland 20878.
Sturm
S
S
Anderson
W F
WF
Eglitis
M A
MA
D016475:3T3 Cells; D000818:Animals; D000911:Antibodies, Monoclonal; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D009052:Leukemia Virus, Murine / Q000378:metabolism*; D051379:Mice; D011991:Receptors, Virus / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D011991:Receptors, Virus