Serological and Structural Heterogeneity of HLA-DR and DQ Antigens in HLA-DR4 Homozygous B Lymphoid Cells

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Titration in the ELISA assay of a panel of anti HLA class II monoclonal antibodies with homozygous HLA-DR4 B lymphoid cell lines showed that the monoclonal antibody Q2/80 displays a low titer with the cell lines expressing the HLADw15 specificity, a high titer with the cell lines expressing the HLA-DKT2 specificity and an intermediate titer with the cell lines expressing the HLA-Dw4, Dw10 and Dw14 specificities. Because of the limited and somehow conflicting information about the structural heterogeneity of HLA-DR and DQ antigens isolated from HLA-DR4 homozygous lymphoid cells expressing different HLA-D allospecificities, HLA-DR and DQ antigens were analysed by 2-dimensional gel electrophoresis (either IEF or NEPHGE in the first dimension and SDS-PAGE in the second dimensions) utilizing some of the cell lines and monoclonal antibodies utilized by other investigators. Heterogeneity was detected in the β chains but not in the α chains of HLA-DR and DQ antigens isolated from the various cell lines. Specifically the two β chains of HLA-DR antigens associated with HLA-Dw15 (referred to as β1 and β2) are more basic than those associated with HLA-DKT2 (referred to as β3 and β4). The latter two are more basic than the two β chains associated with HLA-Dw10 (referred to as β5 and β6). The two β chains associated with HLA-Dw4 and with HLA-Dw14 do not appear to differ from each other; one has the same electrophoretic mobility as β2 and the other one as β3. The HLA-D associated heterogeneity of β chains of HLA-DR antigens resides in their polypeptide moiety, since it was detected in antigens synthesized by B lymphoid cells in presence of tunicamycin as well as following removal of carbohydrates with enzymes. The electrophoretic heterogeneity of the β chains of HLA-DQ antigens was less marked than that of HLA-DR antigens, since the β chain associated with HLA-Dw15 (referred to as Qβ1) was more basic than those associated with HLA-DKT2, Dw10 and Dw14 (referred to as QB2), which did not differ from each other. The β chain associated with HLA-Dw4 consisted of both Qβ1 and Qβ2. The present study has shown for the first time serological heterogeneity among HLA-DR4 alloantigens associated with different HLA-D allospecificities. Furthermore our results suggest that the conflicting results in the literature about the structural heterogeneity of HLA-DR4 antigens may be caused by various labelling procedures and/or by the sensitivity of the 2-D electrophoretic procedure and that HLA-DR4 lymphoid cell lines with the same HLA-D allospecificity may differ in the antigenic profile or in the expression of HLA-DQ antigens.
北里大学の論文
1987-06-30

Serological and Structural Heterogeneity of HLA-DR and DQ Antigens in HLA-DR4 Homozygous B Lymphoid Cells

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