Glycolytic regulation of rheumatoid factor autoreactive AM14 B cell activation and plasma cell differentiation 2507

Abstract Description Multiple evidence suggests that an elevated glucose metabolism in T cells and B cells contributes to lupus pathophysiology. Autoantigen-specific AM14 B cells but not foreign-Ag specific B cells showed a reduced activation and differentiation into antibody producing cells in mice treated with an inhibitor of glycolysis. To study the intrinsic role of glycolysis, we have developed a CRISPR/Cas9 method to knock down the glucose transporter Glut1 in Ag-specific B cells. The knockout efficiency was validated by flow cytometry and qRT-PCR. Glut1KO AM14 B cells and control AM14 B cells transferred into BALB/c recipients were stimulated with the anti-chromatin IgG2aa hybridoma PL2-3. The frequency of total AM14 B cells, AM14 plasmablasts and plasma cells decreased in the Glut1KO transfer group, while IgD?Ki67?% AM14 B cells remained unchanged, indicating that glycolysis does not play a role in autoreactive B cell proliferation. In addition, the production of 4-44 IgM (AM14 idiotype) decreased in the Glut1KO transfer group. Finally, CD80 and CD86 expression on AM14 B cells decreased in the Glut1KO group. These findings suggest that glycolysis has a selective intrinsic impact on the activation and co-stimulatory capacity of autoreactive B cells. The same approach is in progress with NP-specific B1-8 B cells to assess for the first time the glycolysis requirement of Ag-specific autoreactive B cells in the context of lupus. Funding Sources R37AI128901 Topic Categories Basic Autoimmunity (BA)