Figure 5. Different subsets of cLPL cells produce either IL-17 or IL-22 after restimulation.
(A) Dot plots of the three major RORγt expressing populations that were identified from bulk cLPL cells isolated after 3 days of DSS. Intracellular cytokine analysis of cLPL cells showing IL-17 vs IL-22 production for each population after 3 hrs of restimulation in vitro. Representative dot plots are shown. (B) Percentage of RORγt-GFP+ cells expressing either IL-17 or IL-22 by population, combined data from two experiments is shown, means ± SD. (C) Detection of intracellular IL-17 (x-axis) and IFNγ (y-axis) in cLPL from either naïve or day 3-DSS treated mice after 3–4 hours restimulation ex vivo. Representative zebra plots show either a CD4+ T cell gate (top two rows) or a γδ+ T cell gate (bottom two rows). (D) Bar graph showing percentage of either CD4+ or γδ T cells from DSS treated mice that are positive for IFNγ (top) and IL-17 (bottom), combined data from 2 experiments, with means ± SEM (*p<0.0001; **p<0.04). Two-tailed, Students t test performed. (E) Colon LPL cells from Tcrd +/− and Tcrd −/− mice gated on live, CD3+Thy1+ cells. (F) Colon LPL cells as in (E), stimulated with PMA/ionomycin and gated on the populations indicated. (G) Detection of serum FITC dextran after 3 days of 2.5% DSS in drinking water in the indicated mice.