Autoantibodies secreted by TLR7-stimulated B cell subsets from mice with primary Sjogren's syndrome

Spleens (n = 2-3) were harvested from mice with primary Sjogren's syndrome (NOD.B10) and cells were sort-purified. Follicular B cells (FO) (6 million), Marginal zone B cells (4.5 million) and age-associated B cells (500,000) were sorted. Cells were cultured in 200 uL of complete RPMI media containing 62.5 ng/mL of imiquimod, a TLR7 agonist. Cells were cultured for 6 days, and the supernatants were harvested and stored at -20 C. Samples were shipped to UT Southwestern for autoantigen array analysis. 5 samples were analyzed for each subset. Each sample was derived from 2-3 pooled spleens from aged NOD.B10 females. Cells were sort-purified using the following markers: FO: B220+, CD23+, CD21lo/-, MZ: B220+, CD23-, CD21+, ABC: B220+, CD11c+, CD11b+.