Panel A shows the percentage and number of IGRP206–214 reactive CD8 T cells in the spleen of 7–9-week-old Cre-positive Tnfrsf9-positive-positive (wild-type) and Cre-positive Tnfrsf9-flox-flox (knockout) female mice. The flow cytometry profiles on the left display viable cells with CD3 and CD8 markers, and CD44 and Major Histocompatibility Complex Class I tetramer markers. The bar graphs on the right summarize the percentage and number of these cells, indicating a significant increase in knockout mice. Panel B presents similar data for the pancreatic lymph node, with flow cytometry profiles and bar graphs showing a significant increase in IGRP206–214 reactive CD8 T cells in knockout mice. Panel C illustrates the proliferation of adoptively transferred NY8.3 CD8 T cells in the pancreatic lymph nodes of 7–9-week-old Cre-positive Tnfrsf9-positive-positive (wild-type) and Cre-positive Tnfrsf9-flox-flox (knockout) male mice. The flow cytometry profiles on the left show viable cells with CD3 and CD8 markers, and Carboxyfluorescein Succinimidyl Ester labeling. The bar graph on the right summarizes the percentage of divided NY8.3 CD8 T cells, showing no significant difference between wild-type and knockout mice.
Increased accumulation of IGRP 206-214 -specific CD8 T cells in the spleen and PLN of Cre + -Tnfrsf9 fl/fl mice, related to Fig. 2. (A and B) Frequencies and absolute number of IGRP206–214-reactive CD8 T cells in the spleen (A) and PLN (B) of 7- to 9-wk-old Cre+-Tnfrsf9+/+ (WT) and Cre+-Tnfrsf9fl/fl (KO) female mice. The representative flow cytometry profiles (left) and the summarized results (right) from four experiments are shown. *P < 0.05; **P < 0.01 by an unpaired t test. (C) Proliferation of adoptively transferred NY8.3 CD8 T cells in the PLNs of 7- to 9-wk-old Cre+-Tnfrsf9+/+ (WT) and Cre+-Tnfrsf9fl/fl (KO) male mice. Splenocytes isolated from 7- to 9-wk-old NOD.Rag1−/−.NY8.3 mice were labeled with CFSE and transferred into WT or KO mice. The gating strategy (left) and combined results (right) from five experiments are presented. ns: not significant by an unpaired t test.