Effects of CsA on murine BMDC differentiation, allostimulatory capacity, and cytokine production. (A) CsA does not affect DC differentiation. Day 3 DCs were stimulated with 1 μg/mL CsA for 72 hours. During the last 24 hours of CsA treatment, DCs were matured with 100 ng/mL LPS. DCs were then collected and analyzed for expression of major histocompatibility complex (MHC) (Ia) and costimulatory molecules (CD80, CD86, and CD40). Untreated day-7 DCs were used as a control (DC). Unfilled histograms depict isotype control, and shaded histograms represent surface expression level of developmental markers. (B) CsA inhibits the allostimulatory capacity of DCs. MLR was performed as described in “Materials and methods.” DC (day-7 DCs), DC-CsA (day-5 DCs treated with 1 μg/mL CsA alone for 48 hours), DC-LPS (day-6 DCs stimulated with 100 ng/mL LPS alone for 24 hours), and DC-CsA/LPS (day-5 DCs stimulated with both 1 μg/mL CsA and 100 ng/mL LPS) were used in this assay at various stimulator-responder (BALB/c splenic T cells) ratios, as indicated. (C) CsA inhibits TNF-α and IL-12p70 production but augments IL-10 production. Day 5 DCs were treated as indicated and were analyzed for cytokine production by ELISA. CsA concentration in CsA/LPS samples was 1 μg/mL. Results are expressed as means ± SEM (B-C).