Figure 3. Activation of β-Cell-Specific T Cells in NOD Versus NOD.Il27^−/− Recipients.

(A and B) Purified splenic BDC2.5 CD4 T cells (2 × 10⁶) were labeled with eFluor670 proliferation dye and transferred into 8- to 9-week-old female NOD or NOD.Il27^−/− recipients. PLNs were harvested 5 days post-transfer, and proliferation (A) and CD44 and CD62L expression (B) of CD4⁺eFluor670⁺ cells was analyzed using flow cytometry.

(C and D) Purified splenic NY8.3 CD8 T cells (3 × 10⁶ to 5 × 10⁶) were labeled with eFluor670 proliferation dye and transferred into sex-matched 7- to 9-week-old NOD or NOD.Il27^−/− recipients. PLNs were harvested 5 days post-transfer, and proliferation (C) and CD44 and CD62L expression (D) of CD8⁺eFluor670⁺ cells were analyzed using flow cytometry.

(E and F) Purified splenic CD45.2⁺ NY8.3 CD8 T cells (5 × 10⁶) were transferred into 7- to 9-week-old female NOD or NOD.Il27^−/− recipients (expressing CD45.1). Sixteen hours after transfer, PLNs were harvested and analyzed for CD69 (E) and CD44 (F) expression on CD45.2⁺CD8⁺NRP-V7 tetramer⁺ cells using flow cytometry.

Representative flow cytometry profiles are shown in the left panels, and summarized results are presented in the right panels. Summarized results are from two or three independent experiments. Each symbol represents a mouse. The horizontal bar depicts the mean. *p < 0.05 and **p < 0.01 by Mann-Whitney test. NS, not significant.