Serological analysis of the dissociation process of HLA-B and C class I molecules.

Two forms of HLA class I molecules reacting differentially with the HC-10 monoclonal antibody were identified at the surface of HLA-A3, B7, Cw3 or Cw7 human cells. The HC-10-nonreactive form (which includes all HLA-A3 and a large fraction of HLA-B7, Cw3 and or Cw7 molecules) corresponds to heavy chains apparently tightly associated to beta 2-microglobulin. The HC-10-reactive form (which represents only a fraction of cell surface expressed HLA-B7, Cw3 and Cw7 molecules) corresponds to heavy chains loosely but still associated to beta 2-microglobulin. Further biochemical analyses and the study of mouse transfected cells expressing other HLA class I specificities led to the following conclusions: (a) dissociation of HLA-B and C molecules is a multistep phenomenon, the various stages being identifiable serologically; (b) acquisition of the HC-10 antigenic determinant appears as a hallmark of HLA class I molecules engaged in the process of dissociation; however, its expression does not imply complete separation of heavy and light chains; (c) only the initial stage of the dissociation process can be identified on cell surfaces, whereas (d) following addition of detergent, dissociation of HLA-B and C molecules spontaneously proceeds further, resulting in accumulation in cell lysate of cell surface-derived isolated HLA-B and C class I heavy chains.

Human beta 2-microglobulin specifically enhances cell-surface expression of HLA class I molecules in transfected murine cells.

Sequential transfections of P815 murine mastocytoma cells with class I gene encoding either HLA-Cw3, HLA-A3, or HLA-B7 H chain and subsequently with a human beta 2-microglobulin gene were performed to evaluate the relative efficiency of human and murine beta 2-microglobulins in promoting the cell-surface expression of HLA-class I molecules. A 6-, 11-, and 40-fold specific enhancement of the cell-surface expression of HLA-Cw3, HLA-A3, and HLA-B7 molecules, respectively, was observed in cells co-transfected with human beta 2-microglobulin gene. This effect was attributed to a more efficient association of HLA H chains with human than with murine beta 2-microglobulin, which apparently allowed a more rapid transport of the HLA molecules from the endoplasmic reticulum to the Golgi apparatus.

[Contribution au dosage du glycerol dans les glycerines techniques--II]. / Contribution au dosage du glycerol dans les glycerines techniques--II.

The authors confirm experimentally the hypothesis of the volatilization of formic acid during the volumetric determination of glycerol by the periodate method. The reproducibility of the results can be improved if sodium formate is added.