Abstract
The pathogenesis of autoimmune diseases has not been completely elucidated yet, and only a few specific treatments have been developed so far. In autoimmune diseases mediated by pathogenic autoantibodies, such as systemic lupus erythematosus, the specific detection and analysis of autoreactive B cells is crucial for a better understanding of the physiopathology. Biological characterization of these cells may help to define new therapeutic targets. Very few techniques allowing the precise detection of autoreactive B cells have been described so far. Herein we propose a new flow cytometry technique for specific detection of anti-nucleosome B cells, which secrete autoantibodies in systemic lupus erythematosus, using labeled nucleosomes. We produced different fluorochrome-labeled nucleosomes, characterized them, and finally tested them in flow cytometry. Nucleosomes labeled via the cysteines present in H3 histone specifically bind to autoreactive B cells in the anti-DNA transgenic B6.56R mice model. The present work validates the use of fluorochrome-labeled nucleosomes via cysteines to identify anti-nucleosome B cells and offers new opportunities for the description of autoreactive B cell phenotype.
Highlights
IntroductionMany autoimmune diseases, such as systemic lupus erythematosus (SLE), are characterized by the presence of B cells that are directed against self antigens (i.e. autoreactive B cells) and produce autoantibodies (autoAbs)[1]
Many autoimmune diseases, such as systemic lupus erythematosus (SLE), are characterized by the presence of B cells that are directed against self antigens and produce autoantibodies[1]
In order to develop a new flow cytometric method to detect autoreactive B cells, we chose the nucleosome as an autoantigen
Summary
Many autoimmune diseases, such as systemic lupus erythematosus (SLE), are characterized by the presence of B cells that are directed against self antigens (i.e. autoreactive B cells) and produce autoantibodies (autoAbs)[1]. Anti-nucleosome antibodies are part of a large family of antibodies directed against epitopes of histones, dsDNA or conformational epitopes created by the interactions between dsDNA and histones[8] They may precede the clinical development of SLE up to 10 years[4], and as anti-DNA antibodies, they are SLE-specific and associated with the disease activity[9]. These autoantibodies form immune complexes within blood vessels and kidneys leading to chronic inflammation, and play a critical role in the pathogenesis[6, 10,11,12]. The present work opens up the use of cysteine labeled nucleosomes to identify and characterize anti-nucleosome B cells for a better understanding of SLE physiopathology
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