Figure 4.

Regulation of the Effector T Cell Response by Antigen Receptor Signaling

(A) Ly5.1 Nur77-GFP OT-I CD8 T cells were adoptively transferred into congenic Ly5.2 C57BL/6 mice and vaccinated IN next day with OVA protein formulated in the indicated adjuvants. At days 2, 5, or 8 PV, cells from lymph nodes and lungs were stained with K^b/SIINFEKL tetramers, anti-Ly5.1, anti-Ly5.2, anti-CD8, and anti-CD44 antibodies. The GFP MFIs in donor Ly5.1^+ve OT-I CD8 T cells were quantified by flow cytometry.

(B) Wild-type non-transgenic (WT) and transgenic KLF2-GFP mice were vaccinated with NP protein formulated in adjuvants, as in (A). At day 8 PV, lung cells were stained with anti-CD8, anti-CD44, and D^b/NP366 tetramers. The overlay histogram shows GFP fluorescence (MFI) for the gated tetramer-binding CD8 T cells from WT (black) and KLF2-GFP transgenic (red) mice.

(C) B6 mice were vaccinated with NP protein formulated in various adjuvants, as in (A). At day 8, PV lung cells were stained with anti-CD8, anti-CD44, anti-PD-1, and D^b/NP366 tetramers. Plots show the percentages of PD-1^+ve cells among the gated D^b/NP366 tetramer-binding CD8 T cells.

(D) Statistical correlation analysis between the percentages of PD-1^+ve CD8 T cells and the percentages of tetramer^+ve CD8 T cells at day 8 PV.

Data are pooled from two independent experiments or represent one of two independent experiments. Comparisons were made using one-way ANOVA test with Tukey-corrected multiple comparisons. For (D), we used two-way ANOVA, Student’s t test, and simple regression analysis; ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.