Stable transformation of mouse L cells for human membrane T-cell differentiation antigens, HLA and beta 2-microglobulin: selection by fluorescence-activated cell sorting.
We isolated stable transformants of mouse L cells expressing human cell surface differentiation antigens by using immunofluorescence with monoclonal antibodies and selection with a fluorescence-activated cell sorter (FACS). Mouse L cells (TK-) were cotransformed with human cellular DNA and the herpes simplex virus thymidine kinase (TK) gene. TK+ transformants were first selected. The TK+ populations were stained with various fluorescent antibodies to membrane antigens, and positive cells were sorted and cloned by using a FACS. Transformants for HLA class I antigens, for beta 2-microglobulin, and for the T-cell differentiation antigens Leu-1 and Leu-2 were isolated. The frequency of antigen transformants among the TK+ transformants was about 0.5 X 10(-3). The sizes of the HLA, Leu-1, and Leu-2 molecules expressed by the transformants were the same as those of the proteins present on DNA-donor cells.
D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte; D000954:Antigens, Surface / Q000276:immunology*; D001609:Beta-Globulins / Q000276:immunology*; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D005434:Flow Cytometry; D005796:Genes; D006680:HLA Antigens / Q000276:immunology*; D006801:Humans; D007739:L Cells / Q000276:immunology; D007945:Leukemia, Lymphoid / Q000276:immunology; D051379:Mice; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology*; D013937:Thymidine Kinase / Q000235:genetics; D001613:beta 2-Microglobulin / Q000276:immunology*
D000945:Antigens, Differentiation, T-Lymphocyte; D000954:Antigens, Surface; D001609:Beta-Globulins; D006680:HLA Antigens; D001613:beta 2-Microglobulin; D013937:Thymidine Kinase