Probing the stability of class I major histocompatibility complex (MHC) molecules on the surface of human cells

Abstract

 We used binding of a fluorescent adduct of β₂-microglobulin, fluorescein β₂m, 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 β₂m and possibly peptide as well. Hence Fl-β₂m reports on the stability of the class I HLA trimer. A small fraction of HLA molecules, ∼5%, binds Fl-β₂m on both resting and activated T cells. A larger fraction of all HLA molecules binds Fl-β₂m in FO-1 cells, β₂m-deficient cells, transfected with various B2m genes. HLA molecules of FO-1 cells are more stable when expressed with human β₂m, than when expressed with mouse β₂m. The non-covalent association of HLA heavy chains, β₂m and peptide implies that eventually every molecule of HLA trimer ought to dissociate and bind Fl-β₂m. In fact, the extent of exchange is limited by the lifetime of a given molecule at the cell surface. β₂m exchange decreases as cell concentration increases, suggesting that some density-dependent process acts to enhance degradation or denaturation of β₂m-free HLA heavy chains.