Endocytosis of HLA and H-2 molecules on transformed murine cells measured by fluorescence dequenching of liposome-encapsulated carboxyfluorescein.
We have studied internalization of cell surface proteins encoded by genes of the human major histocompatibility complex (HLA) transferred into murine L cells, in comparison with mouse (H-2) histocompatibility determinants. This internalization was measured by the use of monoclonal antibodies directed at these determinants, to which were bound methotrexate- and carboxyfluorescein-containing liposomes covalently coupled to protein A. In addition to the effect of methotrexate on the cells, the technique of fluorescence self-quenching release was used to measure the kinetics of internalization for single cells using a flow cytofluorometer. The native and foreign gene products were internalized in an apparently identical manner. These techniques can be applied to cells expressing genes with altered or deleted segments thus providing a basis for the analysis of the effect of sequence modifications on the internalization of the encoded molecule.
D000818:Animals; D000911:Antibodies, Monoclonal; D002472:Cell Transformation, Viral; D004705:Endocytosis; D000939:Epitopes / Q000032:analysis; D005452:Fluoresceins; D006183:H-2 Antigens / Q000235:genetics* / Q000276:immunology; D006680:HLA Antigens / Q000235:genetics* / Q000276:immunology; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D007739:L Cells / Q000276:immunology; D008081:Liposomes / Q000008:administration & dosage*; D008285:Major Histocompatibility Complex; D008727:Methotrexate / Q000494:pharmacology; D051379:Mice; D013539:Simian virus 40 / Q000235:genetics; D013050:Spectrometry, Fluorescence
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005452:Fluoresceins; D006183:H-2 Antigens; D006680:HLA Antigens; D008081:Liposomes; C024098:6-carboxyfluorescein; D008727:Methotrexate