Serologic dissection of HLA-D specificities by the use of monoclonal antibodies. II. Distinction between HLA-Dw2 and HLA-Dw12.
Three cytotoxic monoclonal antibodies, HU-11, HU-32, and HU-33, specific for human Ia-like antigens were used to analyze the two HLA-DR2-associated HLA-D specificities, HLA-Dw2 and HLA-Dw12. In the HLA-Dw2, DR2, MB1 homozygous B-cell line EB-CMG, the binding of radiolabeled HU-32 and HU-33 was strongly inhibited by the addition of nonlabeled HU-11, whereas no inhibition occurred in the HLA-Dw12, DR2, MB1 homozygous B-cell line EB-KT. To confirm this differential inhibition pattern further, F(ab')2 fragments were prepared from HU-11, and their ability to inhibit complement-dependent lysis mediated by HU-32 and HU-33 was assessed against a total of five homozygous typing cell lines homozygous for HLA-Dw2, DR2, MB1 or HLA-Dw12, DR2, MB1, including EB-CMG and EB-KT. Here again, the same differential inhibition pattern as that observed in the radiobinding inhibition assays was obtained. Thus, the data suggest that the two kinds of HLA-DR2-positive homozygous typing cell lines with distinct HLA-D specificity can be distinguished from each other by using solely serologic methods. This is the first clear-cut serologic distinction made between homozygous typing cells defining HLA-Dw2 and those defining HLA-Dw12, since no serologic means that enables one to distinguish one from the other has been available.
D000911:Antibodies, Monoclonal; D001402:B-Lymphocytes / Q000276:immunology; D002460:Cell Line; D003602:Cytotoxicity, Immunologic; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis; D005802:Genes, MHC Class II; D006681:HLA-D Antigens; D015802:HLA-DR2 Antigen; D006720:Homozygote; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000032:analysis; D010641:Phenotype
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006681:HLA-D Antigens; D015802:HLA-DR2 Antigen; C039901:HLA-Dw12 antigens; D007140:Immunoglobulin Fab Fragments