Figure 6: Ectopic expression of NFIL3 in CD4⁺ T cells attenuates gut pathology in adoptively transferred enteritis/colitis.

Naïve CD4⁺ T cells from C57BL/6 mice were transduced with NFIL3-expressing retrovirus (NFIL3) or control empty retrovirus (GFP). Cells were injected intraperitoneally (i.p.) into Rag1^−/− recipient mice to induce gut inflammation (0.7 × 10⁷ cells per mouse). (a) Wasting disease was monitored for 10 weeks after transfer. Statistical analysis was performed on the combination of total animals from two independent experiments. Data are shown as mean±s.e.m. Mann–Whitney test two-tailed P=0.0064. (b) Immunopathology of gut inflammation. Two randomly selected tissue sections from each animal were graded for crypt damage and T-cell infiltration. Two scores were given to each animal to represent the overall gut immunopathology. Data are shown as mean±s.e.m. Unpaired t-test two-tailed P=0.0056. (c) The recipient mice were killed 6 weeks after T-cell transfer for ex vivo analysis of cytokine production (IL-2, IL-10 and IFN-γ) and Tim-3 expression by flow cytometry. (d) Naïve CD4⁺ T cells from WT FoxP3-GFP KI and NFIL3^−/− x FoxP3-GFP KI mice (CD4⁺CD62L⁺GFP⁻) were differentiated into Th1 cells with or without the presence of IL-27. On day 6, GFP⁺ (FoxP3⁺) cells were isolated by cell sorting and were injected i.p. into Rag1^−/− recipients to induce colitis. Colon pathology was analysed 8 weeks after transfer. Each score represents the pathology from one recipient mouse (NFIL3^−/− Th1 n=8; NFIL3^−/− Th1+IL-27 n=7; WT Th1 n=7; WT Th1+IL-27 n=9. Data are shown as mean±s.e.m., t-test).