Extended Data Figure 3. Extended characterization of H. hepaticus-specific TCRtg cell differentiation.
a, HH7-2tg;Rag1−/− mice do not develop Treg cells in the thymus. Representative flow cytometry plots of Treg (Foxp3+CD25+) frequency in indicated tissues of H. hepaticus-free HH7-2tg;Rag1+/− (n=3) or HH7-2tg;Rag1−/− (n=3) mice. b, c, HH7-2tg Rag1−/− and Rag1+/− donor-derived T cells differentiated into equal frequencies of RORγt+ Treg in the LI of WT mice. Equal numbers (2,000) of congenic isotype-labeled HH7-2tg Rag1+/− (CD45.1/45.1) and Rag1−/− (CD45.1/45.2) naïve T cells were co-transferred into H. hepaticus-colonized WT B6 mice. Cells from the LILP were analyzed two weeks after transfer. Data summarize two independent experiments (n=6). b, Representative flow cytometry plots of donor and recipient T cell frequency (left), and RORγt and Foxp3 expression (right) (n=6). c, Frequencies of Treg (Foxp3+), TH17 (Foxp3−RORγt+) and TFH (Bcl6+CXCR5+) cells among HH7-2tg Rag1+/− (n=6) and Rag1−/− (n=6) donor-derived T cells. d, e, 2,000 naïve HH5-1tg cells (CD45.1/45.2) were adoptively transferred into WT B6 mice (CD45.2/45.2) colonized with H. hepaticus. Cells from LILP and CP were analyzed two weeks after transfer. d, Representative flow cytometry plots are shown for RORγt, Foxp3, Bcl6 and CXCR5 expression in donor-derived and recipient CD4+ T cells in indicated tissues. e, Frequencies of Treg (Foxp3+), TH17 (Foxp3−RORγt+) and TFH (Bcl6+CXCR5+) among HH5-1tg donor T cells (n=8). Data are a summary of eight mice from two independent experiments. All statistics were calculated by unpaired two-sided Welch’s t-test. Error bars: mean ± 1 SD. P values are indicated in the figure.