Serological expression after sequential double transfection with purified HLA-A11 gene of mouse fibroblasts carrying human beta-2 microglobulin.
A genomic cosmid library constructed from DNA from a genotyped individual (JF = HLA-A11, Cw-, B38/A26, Cw7, B51) was screened for clones containing class I histocompatibility genes. Among these clones, one was found to carry a 4.8 kb Hind III fragment which is highly correlated with HLA-A11. This clone was used to transfect LMTK+ cultured mouse fibroblast transformants expressing human beta-2 microglobulin. The human beta-2 microglobulin heavy chain-associated determinant was positively detected by the M18 monoclonal antibody. HLA-A11 expression on these doubly transformed cells was specifically demonstrated by complement-dependent cytotoxicity with HLA-A11 + A3-specific but not with HLA-A3-specific monoclonal antibodies. Absorption studies with human alloantisera confirmed the presence on these cells of HLA-A11 determinants and of cross-reacting determinants which absorbed anti-HLA-A1 and -A3 alloantisera. The JF5-J27 transfected cell expressed both heavy and light chains of human class I histocompatibility genes.
D000818:Animals; D000911:Antibodies, Monoclonal; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D006680:HLA Antigens / Q000235:genetics* / Q000276:immunology; D015234:HLA-A Antigens; D006801:Humans; D007518:Isoantibodies / Q000276:immunology; D046911:Macromolecular Substances; D051379:Mice; D014162:Transfection; D001613:beta 2-Microglobulin / Q000235:genetics*
D000911:Antibodies, Monoclonal; D006680:HLA Antigens; D015234:HLA-A Antigens; D007518:Isoantibodies; D046911:Macromolecular Substances; D001613:beta 2-Microglobulin