Identification of autoreactive B cells with labeled nucleosomes.
Vincent GiesAlain WagnerCécile SeifertAurélien GuffroyJean-D FaunyAnne-M KnappJean-L PasqualiThierry MartinHélène DumortierAnne-S KorganowPauline Soulas-SprauelPublished in: Scientific reports (2017)
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.
Keyphrases
- flow cytometry
- systemic lupus erythematosus
- pet imaging
- loop mediated isothermal amplification
- real time pcr
- label free
- induced apoptosis
- disease activity
- dna methylation
- gene expression
- type diabetes
- computed tomography
- circulating tumor
- skeletal muscle
- oxidative stress
- adipose tissue
- positron emission tomography
- signaling pathway
- cell free
- insulin resistance
- quantum dots