Translation and assembly of HLA-DR antigens in Xenopus oocytes injected with mRNA from a human B-cell line.
HLA-DR antigens are polymorphic cell surface glycoproteins, expressed primarily in B lymphocytes and macrophages, which are thought to play an important role in the immune response. Two polypeptide chains, alpha and beta, are associated at the cell surface, and a third chain associates with alpha and beta intracellularly. RNA isolated from the human B-cell line Raji was injected in Xenopus laevis oocytes. Immunoprecipitates of translation products with several monoclonal antibodies revealed the presence of HLA-DR antigens similar to those synthesized in Raji cells. One monoclonal antibody was able to bind the beta chain after dissociation of the three polypeptide chains with detergent. The presence of all three chains was confirmed by two-dimensional gel electrophoresis. The glycosylation pattern of the three chains was identical to that observed in vivo, as evidenced in studies using tunicamycin, an inhibitor of N-linked glycosylation. The presence of alpha chains assembled with beta chains in equimolar ratio was further demonstrated by amino-terminal sequencing. An RNA fraction enriched for the three mRNAs, encoding alpha, beta, and intracellular chains, was isolated. This translation-assembly system and the availability of monoclonal antibodies make it possible to assay for mRNA encoding specific molecules among the multiple human Ia-like antigens.
D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D002051:Burkitt Lymphoma; D002460:Cell Line; D005260:Female; D005802:Genes, MHC Class II; D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II; D006801:Humans; D014176:Protein Biosynthesis / Q000187:drug effects; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000235:genetics* / Q000302:isolation & purification; D014415:Tunicamycin / Q000494:pharmacology; D014981:Xenopus
D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II; D012333:RNA, Messenger; D014415:Tunicamycin