Fig. 3
Antigen-recall assay demonstrates robust responses to MOG of lymph node cells from either WT or VPAC1 KO mice after EAE immunization. EAE was induced to WT and VPAC1 KO mice and draining lymph nodes were isolated at the peak of the WT disease (day 14). Cells were cultured for 48 h at 1 × 106 cells/ml in 96-well plates in the presence of OVA or MOG (10 μg/ml). a Proliferation determined by measurement of [3H]-thymidine incorporation. b Cytokine responses in WT vs. VPAC1 KO lymph node cultures. The levels of IFNγ (Th1), IL-17 (Th17), and IL-10 (Th2/Treg) were measured in the supernatants by ELISA. The expression of IL-4 mRNA in the cultured cells was determined by real-time RT-PCR. Results shown are representative of three independent experiments of n = 8 mice/group. *p < 0.05, **p < 0.01, ***p < 0.001, Student’s t test