Because the Th1-dominated IFN-γ-producing CD4+ T-cell response in control B6 mice is replaced by a Th17-dominated IL-17-producing CD4+ T-cell response in mice with combined defects in IL-12 and type I IFN

receptor,30 the relative production of IL-17 and IFN-γ by L. monocytogenes-specific CD4+ T cells in mice with combined defects in IL-21, IL-12 and type I IFN receptor (TKO) was compared with that in DKO mice, IL-21-deficient mice and control B6 mice (Fig. 4). Surprisingly, the additive effect of IL-21 deficiency in mice with combined defects in IL-12 and type I IFN receptor not only did not ablate, but accentuated IL-17 production after stimulation with the L. monocytogenes-specific I-Ab class II peptide LLO189–201 (Fig. 4a,b). Importantly, increased IL-17 production by L. monocytogenes-specific CD4+ Decitabine clinical trial T cells, which occurs with IL-21 deficiency,

was not restricted only to mice with combined defects in IL-12 and type I IFN receptor because despite sharp reductions in the magnitude of IL-17-producing CD4+ T cells, a similar twofold increase in percentage and total number of IL-17-producing L. monocytogenes-specific CD4+ T cells was found for IL-21-deficient mice compared with B6 control mice (Fig. 4a,b). Interestingly, despite the increased production of IL-17 that occurs in the absence of IL-21, the percentage and absolute numbers of IFN-γ-producing Nutlin-3 ic50 CD4+ T cells were not reciprocally reduced in IL-21-deficient compared with control B6 mice (Fig. 4c). Taken together, these results indicate that IL-21, IL-12 and type I IFNs synergize and play additive inhibitory roles in the differentiation of L. monocytogenes-specific IL-17-producing CD4+ T cells. Interleukin-21 therefore plays dramatically opposing roles in Th17 CD4+ T-cell differentiation under infective and non-infective conditions.

To identify the individual and collective roles of IL-21, IL-12 and type I IFNs in priming protective immunity to secondary L. monocytogenes infection, the susceptibility to re-challenge with virulent L. monocytogenes was enumerated for each Sorafenib ic50 group of mice. Thirty days after primary L. monocytogenesΔactA inoculation, groups of B6, IL-21-deficient, DKO and TKO mice were each challenged with 105 CFUs of virulent Lm-OVA.30,32 Compared with naive mice, L. monocytogenesΔactA-primed mice in each group were uniformly highly protected, and by day 3 after re-challenge contained four to five log10 reductions in recoverable L. monocytogenes CFUs (Fig. 5a). Moreover, by day 5 after re-challenge, virulent L. monocytogenes was cleared from both the spleen and liver in L. monocytogenesΔactA-primed mice in each group. The marked reductions in bacterial burden after re-challenge in L. monocytogenesΔactA-primed compared with naive mice in each group was associated with robust secondary expansion of L.