Two HLA-DQ-specific human-human hybridoma antibodies (TrG6;TrC5) define epitopes also expressed by a transcomplementing hybrid DQ molecule (DQw7 alpha/DQw4 beta).
We have generated two IgG human-human hybridoma Abs, TrG6 and TrC5, that define subsets of HLA-DQ. TrG6 combined selectively with lymphoblastoid cell lines that expressed DQw1 or DQw4. By sequential immunoprecipitation, competition with other mAbs of defined specificity for binding to antigen, and experiments where HLA antigens from cell lysates crosslinked pairs of mAbs, it was established that TrG6 bound a DQ-molecule. mAb TrC5 specifically recognized DQw2+DR3+ and DQw7+DR5+ cells. The reaction pattern of TrC5 with HLA-loss mutants indicated that TrC5 bound to DQw2 of the DQw2+DR3+ haplotype. Antigens in lysate from DQw7+DR5+ cells crosslinked TrC5 to the murine mAbs Tü22 (anti-DQ monomorphic) and IVD-12 (anti-DQw7 + DQw8 + DQw9), demonstrating that on these cells the TrC5 epitope is located on DQw7 molecules. Lysates from DQw7+DR5+/DQw4+DRw8+ heterozygous cells crosslinked TrG6 and TrC5, and available evidence indicated that the epitopes defined by these two mAbs were expressed by the transcomplementing DQ-molecule DQw7 alpha/DQw4 beta, where the DQw7 alpha chain specifies epitope TrC5 and the DQw4 beta chain specifies epitope TrG6. Taken together with published nucleotide sequences of DQ alpha and beta genes, our data are consistent with the conclusion that the amino acids at positions 69 and/or 75 of the DQ alpha chain of DQw2+DR3+ and DQw7+DR5+ haplotypes are critical for epitope TrC5. The previously reported human-human hybridoma Ab TrB12 reacts with DQw6, DQw8, and DQw9. The specificity of the murine mAb IIB3 is similar to that of TrB12, but, unlike TrB12, IIB3 also binds DQw4+ cells.
Kolstad
A
A
Institute of Medical Biology, University of Tromsø School of Medicine, Norway.
Johansen
B
B
Hannestad
K
K
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001667:Binding, Competitive; D002460:Cell Line; D000939:Epitopes / Q000276:immunology; D005816:Genetic Complementation Test; D006683:HLA-DQ Antigens / Q000145:classification / Q000235:genetics / Q000276:immunology*; D006801:Humans; D006824:Hybridization, Genetic; D009154:Mutation
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006683:HLA-DQ Antigens