Analysis of antibody production in a mouse model of Sjögren’s Syndrome

Auto-antigen array data show IgM from salivary B cells is enriched for anti-nuclear antigen specificity, including Ro (SSA) and La (SSB). B cells were sort purified from spleen, cervical lymph nodes and submandibular salivary tissue (SMG) tissue of Id3-/- mice. We also sort purified B cells from spleen and cervical lymph nodes of age and gender matched C57BL/6 mice. Cells (1x10^5 /well) were cultured in 5% RPMI 1640, stimulated with LPS (25 μg/mL) for 6 days, and supernatants harvested. Sera from SS mice were harvested by cardiac puncture immediately following euthanasia. Blood was allowed to clot for two hours and centrifuged at 2,000 g for twenty minutes. Serum was removed and stored at -20°C until use. These undiluted supernatant samples and sera were used to perform the autoantigen array.