Fig. 9
GITR signaling increases pS6 levels in antigen-specific OT-II CD4 and OT-I CD8 T cells. a Wild-type CD45.2 C57BL/6 mice received a 1:1 mixture of 1 million each c and d or 25,000 each e and f of GITR+/+ and GITR−/− CD45.1 OT-II isolated from littermates and were infected i.n. the following day with influenza A/PR8-OVAII. b Representative flow plots depicting the initial and the post-infection GITR+/+:GITR−/− ratio. c At day 3 p.i., cells isolated from the mLN were stained directly ex vivo for the level of phosphorylated-ribosomal protein S6 (pS6) with representative staining on the right. d At day 5 p.i., cells isolated from the lung were stained directly ex vivo for the level of pS6 with representative staining on the right. At day 10 p.i., proportions e and total numbers f of GITR+/+ and GITR−/− OT-II were determined in the lung, mLN, and spleen. g, h Naive CD45.1 mice received a 1:1 mixture of 1 million each of GITR+/+ and GITR−/− CD45.2 OT-I isolated from littermates, and were infected i.n. the following day with influenza A/PR8-OVA. h At day 5 p.i., cells isolated from the lung were stained directly ex vivo for the level of pS6 with representative staining on the right. dMFI refers to the MFI for the specific antibody stain minus the FMO control. Each symbol represents an individual mouse, with bars indicating mean ± SEM. Statistical analyses performed using Wilcoxon test. Data in c, d were pooled from two independent experiments with eight mice. Data in e, f were pooled from two independent experiments with 12 mice. Data in h were pooled from two independent experiments with seven mice
