Endocytosis of HLA and H‐2 molecules on transformed murine cells measured by fluorescence dequenching of liposome‐encapsulated carboxyfluorescein

• Published in: The EMBO journal Vol. 2; no. 12; pp. 2285 - 2291
• Main Authors: Truneh, A., Machy, P., Barbet, J., Mishal, Z., Lemonnier, F.A., Leserman, L.D.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group 01.01.1983
• Subjects: Animals; Antibodies, Monoclonal; Biological and medical sciences; Cell physiology; cell surface; Cell Transformation, Viral; Endocytosis; Epitopes - analysis; Fluoresceins; Fundamental and applied biological sciences. Psychology; H-2 Antigens - genetics; H-2 Antigens - immunology; HeLa Cells - metabolism; histocompatibility antigen H-2; histocompatibility antigen HLA; HLA Antigens - genetics; HLA Antigens - immunology; Humans; L cells; L Cells - immunology; Liposomes - administration & dosage; Major Histocompatibility Complex; Methotrexate - pharmacology; Mice; Molecular and cellular biology; Simian virus 40 - genetics; Spectrometry, Fluorescence; Human; HLA-System; Histocompatibility system; Transformed cell; Rodentia; Fluorescence; Liposome; Monoclonal antibody; H-2-System; Cell surface; Fluorescence spectrometry; Antigen; Proteins; Vertebrata; Endocytosis; Mammalia; Mouse; Cytofluorimetry
• ISSN: 0261-4189; 1460-2075
• DOI: 10.1002/j.1460-2075.1983.tb01736.x

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.