1. Five commercially-available antibodies react differentially with allelic forms of human HLA-DR beta chain
- Author
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Miranda Houtman, Anna Dzebisashvili, Anatoly Dubnovitsky, Genadiy Kozhukh, Lars Rönnblom, Lars Klareskog, Vivianne Malmström, and Leonid Padyukov
- Subjects
Peripheral blood mononuclear cells ,Endokrinologi och diabetes ,HLA-DR beta-Chains ,Immunology ,Leukocytes, Mononuclear ,Humans ,Western blot ,HLA-DR beta chain ,Endocrinology and Diabetes ,Molecular Biology ,Alleles ,HLA-DRB1 Chains - 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.
- Published
- 2022
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