Identification of Antibodies to DQβ:DRα Interisotypic Heterodimers in Human Sera

Abstract

Background. HLA class II antigens, DR, DQ, and DP, comprised an α and β chains, which typically combine, within the same isotype, to form the major histocompatibility complex:peptide complex. Interisotypic pairing is not commonly observed. Although reports of DQβ:DRα heterodimers exist, the pairing was reported to be unstable and, therefore, not studied to any extent. Methods. DQβ:DRα single antigens were produced through transfectant cell lines and used to identify and characterize positive reactive human sera by a multiplex bead-based assay. Results. Stable DQβ:DRα transfectants were constructed. Cell surface staining with class II–specific monoclonal antibodies revealed that some DQB1 alleles appear to be more efficient in expressing DQβ:DRα heterodimers. Interestingly, alleles within the same serological group varied in their efficiency of forming dimers on the cell surface. For example, DQβ0601:DRα had the highest transfection and cell membrane expression efficiency among 16 common DQB1 alleles tested. In contrast, DQβ0603:DRα-positive transfectants demonstrated minimal surface expression. Assembly of DQβ0601:DRα was not affected by the presence of a DQα chain. DQβ0601:DRα and DQβ0603:DRα single-antigen beads were used to screen human sera. Positive sera were identified that reacted to the unique epitopes of DQβ0601:DRα protein on the cell surface of the transfectants. Conclusions. Our studies have demonstrated that unique DQβ:DRα heterodimers can be formed and are stably expressed on the cell surface. Such antigenic combinations, presented on single-antigen beads, demonstrated that patient sera can react with such heterodimers. Investigations on the potential clinical roles of antibodies against such interisotypic heterodimers are now possible.