Figure 3. Instrinsic MAVS signaling is not required for Treg proliferation or suppressive function in vitro.

(a) Tregs from either Mavs^−/− mice or WT mice were column-enriched based on CD4+CD25+ expression and stimulated as noted (left) or co-cultured with CD11c+ column enriched DCs (right). Samples were left either naïve or co-cultured with heat-inactivated whole WNV for 3 days. Anti-CD3 and anti-CD28 antibodies were used for polyclonal stimulation in positive control groups. Error bars reflect +/− SEM. (b) Spleens from Mavs^−/− or WT mice were harvested and homogenized. Tregs were column-enriched based on CD4+CD25+ expression. CD4+ (Tconv) cells were column-enriched through negative selection and CFSE labeled to track proliferation. Cells were co-cultured in decreasing Tconv: Treg proportions with irradiated splenocytes to serve as APCs and incubated for 80 hrs. Statistical significance calculated using two-tailed unpaired Student’s t tests.