Figure 6. CD4 T cells engage TNFR and Fas signaling pathways to induce IL-1β mediated systemic inflammation.

(a) qPCR of Il1b mRNA in lysates of splenocytes collected from WT mice 3–4 h post CD3 Ab injection (50μg, i.v.). Data are normalized to Hprt1. Error bars indicate SEM from n=3 technical replicates. Data are representative of three independent experiments. (b) Expression of intracellular pro-IL-1β measured by flow cytometry in WT live,CD11c⁺ splenocytes from (a). Data are representative of 4 independent experiments. Statistical analysis was performed by paired one-tailed Student’s t-test. *p<0.05. (c) Neutrophil infiltration in the SI-LP or (d) spleen as measured by flow cytometry in WT or Il1b^−/− mice 18 h post CD3 Ab injection (20μg, i.p.) Error bars indicate SEM from n=3–4 independent experiments. (e) Neutrophil infiltration in the spleen as measured by flow cytometry 12 h post OVA_323–339 peptide injection (50μg, i.v.) into WT or Il1b^−/− mice that previously received (1 d prior) OT-II T_H17 cells (5×10⁶, i.v.). Error bars indicate SEM from n=4 independent experiments. (f) qPCR of Il1b mRNA in lysates of splenocytes collected from WT or Tnf^−/− mice 3–4 h post CD3 Ab injection (50μg, i.v.). Data are normalized to Hprt1. Error bars indicate SEM from n=3 technical replicates. Data are representative of two independent experiments. (g) Neutrophil infiltration in the spleen as measured by flow cytometry in WT and Tnf^−/− mice 3 h post CD3 Ab injection or (h) Fas^fl/fl and Fas^fl/fl x CD11c-cre (Fas^ΔDC) mice 18 h post CD3 Ab injection (20μg, i.p.). All Ly6G⁺ cells were previously gated on live,CD11b⁺F480^- cells. Error bars indicate SEM from n=4–6 independent experiments. (a, c, d, e, g) Statistical analysis was performed by unpaired, one-tailed Student’s t-test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.