DC-Based Immunotherapy Combined with Low-Dose Methotrexate Effective in the Treatment of Advanced CIA in Mice

Figure 4

Combination therapy with smDCs and MTX enhanced Treg population and inhibited Th1/Th17 immunity with little influence on Th2 immunity. Advanced CIA mice were treated with smDCs, MTX, or smDCs and MTX, and T cell subsets were analyzed on day 77 after first CII injection. (a) Splenocytes and lymph nodes cells were cultured in the presence of CII (50 μg/mL) for 72 h. Treg population and Foxp3 mRNA expression were analyzed by flow cytometry and qRT-PCR, respectively. (b) Single cell suspensions were cultured with 50 μg/mL CII for 72 h. Cytokines secreted from the culture of splenocytes and lymph node cells were assessed. The levels of IFN-γ, IL-17, and IL-10 were measured by ELISA in the culture supernatants. Statistical data are shown from more than 3 different sets of experiments as mean ± SEM. , , in Student’s -test.