Five commercially-available antibodies react differentially with allelic forms of human HLA-DR beta chain. (December 2022)

• Record Type: Journal Article
• Authors: Houtman, Miranda; Dzebisashvili, Anna; Dubnovitsky, Anatoly; Kozhukh, Genadiy; Rönnblom, Lars; Klareskog, Lars; Malmström, Vivianne; Padyukov, Leonid
• Is Part Of: Molecular immunology. Volume 152(2022)
• Page Start: 106
• Page End: 110
• Publication Date: 2022-12
• Subjects: HLA-DR beta chain -- Peripheral blood mononuclear cells -- Western blot; HLA human leukocyte antigen -- PBMC peripheral blood mononuclear cell; Immunochemistry -- Periodicals; Molecular biology -- Periodicals; Immunochemistry -- Periodicals; Allergy and Immunology -- Periodicals; Molecular Biology -- Periodicals; Immunochimie -- Périodiques; Biologie moléculaire -- Périodiques; Immunochemistry; Molecular biology; Periodicals; Electronic journals; 571.96
• DOI: 10.1016/j.molimm.2022.10.008
• Languages: English
• ISSNs: 0161-5890

Abstract

Abstract: Allelic variants of HLA-DRB1 have been associated with a variety of autoimmune and infectious diseases. Although the precise molecular mechanisms by which HLA-DRB1 alleles predispose to a particular disease are currently unclear, it has been shown that mRNA expression levels of HLA-DRB1 are dependent on the different alleles. We aimed to measure HLA-DR beta chain levels in peripheral blood mononuclear cells of individuals carrying HLA-DRB1 *03:01/*04:01 and HLA-DRB1 *03:01/*15:01 alleles by western blotting, using five commercially-available HLA-DRB antibodies. We observed highly heterogeneous binding of the tested antibodies to the different allelic forms of the HLA-DR beta chain. Overall, we show that current immunological research that employs available antibodies to detect HLA-DR beta chains is biased towards detection of specific variants of the protein; this may cause significant discrepancy in quantification of protein expression in a heterogeneous human population. Highlights: RNA expression levels of HLA-DRB1 are dependent on the different HLA-DRB1 alleles. Tested commercial HLA-DRB antibodies detect particular allelic forms of HLA-DRB1. Novel tools are needed to study allele-specific protein expression of HLA-DRB1.