Dual parameter, quantitative cytofluorometric analysis of endogenous H-2Kk and foreign HLA class I molecules expressed at the surface of murine transformed L cells.
By using a calibrated dual laser cell sorter and monoclonal antibodies directly conjugated to fluorescein and rhodamine and specific for H-2Kk and HLA class I antigens, quantitative cytofluorometric analysis was performed on individual HLA-A3 or -CW3 transformed mouse L cells (H-2k). More than 80% of these cells expressed both HLA class I and H-2Kk molecules. Their respective levels of expression were calculated: a mean of 4 X 10(5) HLA class I and 2.3 X 10(5) H-2Kk molecules per single cell. Quantitative comparison with control untransformed L cells and double fluorescence contour maps showed a positive correlation between the levels of expression of HLA class I and H-2Kk molecules suggesting that expression of foreign class I molecules did not occur at the expense of the endogenous H-2k product.
D000818:Animals; D000911:Antibodies, Monoclonal; D002469:Cell Separation; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D006183:H-2 Antigens / Q000032:analysis* / Q000276:immunology; D006680:HLA Antigens / Q000032:analysis* / Q000276:immunology; D007739:L Cells; D051379:Mice; D012235:Rhodamines; D013194:Staining and Labeling; D013861:Thiocyanates; D014170:Transformation, Genetic
D000911:Antibodies, Monoclonal; D005452:Fluoresceins; D006183:H-2 Antigens; C024755:H-2K(K) antigen; D006680:HLA Antigens; D012235:Rhodamines; D013861:Thiocyanates; D016650:Fluorescein-5-isothiocyanate