Figure 2. Adoptively transferred Th17 cells evolve in vivo into a distinct Th1-like population.
TRP-1 CD4+ T cells (Thy1.2+) were cultured under Th1 and Th17 conditions for 7 days. Serial gene expression profiling was performed on cells preserved on the day of adoptive transfer (day 0) and on highly purified cells recovered on day 5 and 15 from spleens and lymph nodes 5Gy irradiated B6.PL (Thy1.1+) animals (2–4 independent replicates/condition for days 5 and 15)
(A) Number of genes 2-fold up- or down-regulated (FDR<0.05 for days 5 and 15) between Th17 and Th1-polarized populations on indicated days is shown in their resting state and upon 2 hour re-stimulation in vitro.
(B) Fold differences in expression of indicated genes as measured by microarray on indicated days, red font indicates a fold difference <2 on day 15.
(C) Heat map of the genes differentially expressed by Th17 and Th1-derived cells recovered ex vivo on day 15 (FC>2, FDR<0.05).
(D) Relative log2 expression of selected differentially expressed genes encoding phenotypic markers of terminal differentiation and end-effector function in Th17 and Th1-derived cells recovered on day 15 after adoptive transfer.
(E) Relative log2 expression of selected differentially-expressed genes correlating with in vivo functionality in Th17 and Th1-derived cells recovered at day 15. Error bars for (D) and (E) represent SEM (n=2–4). List of differentially expressed genes at day 0, 5 and 15 is shown in File S1. See also Figure S2.
(F)(G) Expression of CD62L, CD27, KLRG1 and CCR7 on surviving Th17 or Th1-derived cells. Persisting Thy1.2+ cells in spleens of B6.PL (Thy1.1+) mice treated with Th17 or Th1 polarized cells were analyzed by flow cytometry on day 18 after adoptive transfer. Representative dot plots for indicated phenotypic markers are shown. Bar graphs depict frequency of positive cells in Th1 and Th17 populations. Error bars represent SEM (n=3), (*=p<0.05, **=p<0.01, ***=p<0.001).