Fig. 6
IL-27 and type I IFNs differentially regulate IL-10+Tfh development. a–c 10BiT-Il21-RFP mice were infected with LCMV Cl13 and treated with either isotype control or α-IL-27p28 blocking antibodies on days 3 and 9 p.i. a, b Representative flow plots (a) and summary data (b) depicting the proportion and total number of splenic Thy1.1 (IL-10)+ GP66:I-Ab-specific Tfh cells from control and α-IL-27p28-treated mice on day 14 p.i. The relative expression (gMFI) of IL-10 (Thy1.1) in Tfh cells from experimental mice is also depicted in (b). c Summary graph showing serum levels of LCMV-specific IgG1 and IgG2a on day 21 p.i. d Naive CD4 T cells were purified and cultured under Th0 or Tfh-polarizing conditions in the presence or absence of recombinant IL-27. Flow plots (left) and summary data (right) showing the proportion of Thy1.1 (IL-10+) IL-21-tRFP+ cells after 3 days of in vitro culture. e WT mice were infected with LCMV Arm or LCMV Cl13. (e) Summary graph showing serum levels of IL-27 in experimental mice on day 7 p.i. f, g 10BiT-Il21-RFP mice were infected with LCMV Cl13 and treated with either isotype control or α-IFNAR blocking antibodies on days 10, 14, and 18 p.i. f, g Representative flow plots (f) and summary data (g) depicting the proportion and total number of splenic Thy1.1 (IL-10)+ CXCR5+PD-1+ GP66:I-Ab-specific Tfh cells from control and α-IFNAR-treated mice on day 21 p.i. The relative expression of IL-10 (Thy1.1) in Tfh cells from experimental mice is also shown in g. Summary data (box and whisker plots showing median (midline) and interquartile range with upper and lower whiskers representing the range of data distribution in b, c, and g or mean + /- SEM in d, e) are from 3 to 5 mice/group and are representative of two to three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.0001. Data were analyzed using two-tailed unpaired student’s t tests. See also Figure S7
