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. 2014 Feb 13;63(3):1008–1020. doi: 10.2337/db13-0859

Figure 6.

Figure 6

EXO injections induce systemic inflammatory and/or autoimmune responses in young NOD mice. A: NOD mice (1 week old) were injected intraperitoneally three times (waiting for 7 days between injections) with either MIN6-EXOs or NOD-EXOs at 5 µg per animal per injection (2.5 µg for the first injection) or left untreated (Control). Cells were collected at the age of 4 weeks from PanLNs or iLNs and were cultured ex vivo for 72 h without EXO stimulation. Release of cytokines and chemokines was measured by CBA assay. B: Spleen cells from the control or treated mice were stimulated with anti-CD3/28 beads for 3 h in the presence of Brefeldin A (10 µg/mL), followed by CD4 and intracellular IFN-γ staining. C: Splenocytes from the 4-week-old treated or control mice (three mice per group) were cultured for 72 h without adding EXOs. IFN-γ concentration in the culture supernatants was determined by CBA assay. The experiment was repeated once with similar observations. D: CFSE-labeled splenocytes from the treated or control mice were stimulated with iMSC EXOs for 72 h. Proliferating B cells were identified as B220+/CFSE-lo. EXOinj, EXO-treated; TNF-α, tumor necrosis factor-α.