Probing the stability of class I major histocompatibility complex (MHC) molecules on the surface of human cells.
We used binding of a fluorescent adduct of beta2-microglobulin, fluorescein beta2m, to probe the stability of class I HLA molecules on the surface of human cells. The weight of the literature suggests that this ligand binds to heavy chains that have lost beta2m and possibly peptide as well. Hence Fl-beta2m reports on the stability of the class I HLA trimer. A small fraction of HLA molecules, approximately 5%, binds Fl-beta2m on both resting and activated T cells. A larger fraction of all HLA molecules binds Fl-beta2m in FO-1 cells, beta2m-deficient cells, transfected with various B2m genes. HLA molecules of FO-1 cells are more stable when expressed with human beta2m, than when expressed with mouse beta2m. The non-covalent association of HLA heavy chains, beta2m and peptide implies that eventually every molecule of HLA trimer ought to dissociate and bind Fl-beta2m. In fact, the extent of exchange is limited by the lifetime of a given molecule at the cell surface. beta2m exchange decreases as cell concentration increases, suggesting that some density-dependent process acts to enhance degradation or denaturation of beta2m-free HLA heavy chains.
Edidin
M
M
Department of Biology, The Johns Hopkins University, Baltimore, Maryland 21218, USA.
Achilles
S
S
Zeff
R
R
Wei
T
T
D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D004355:Drug Stability; D005452:Fluoresceins; D006680:HLA Antigens / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism; D008545:Melanoma; D051379:Mice; D013601:T-Lymphocytes / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D001613:beta 2-Microglobulin / Q000378:metabolism
D005452:Fluoresceins; D006680:HLA Antigens; D015395:Histocompatibility Antigens Class I; D007140:Immunoglobulin Fab Fragments; D001613:beta 2-Microglobulin