Figure 3.
Hypofunctional CD8 T cells in parous mice share defining features of exhaustion but not anergy. (A) Enrichment of genes up- and down-regulated in T cell exhaustion (chronic viral infection; (Bengsch et al., 2018) in parous CD8 T cells. GSEA was performed using a gene expression dataset generated from bulk RNA-seq of OT-1 T cells sorted from naiveOT-1, pOVAOT-1, and gOVAOT-1 mice. FDR, false discovery rate; NES, normalized enrichment score. Select leading edge genes are depicted. (B) Summary of GSEA performed between OT-1 T cell expression dataset and anergic and exhausted gene sets. Asterisks indicate FDR < 0.05. Eff, T effector cells; TILS, tumor-infiltrating lymphocytes. (C) IR expression on OT-1 cells. OT-1 cells from naiveOT-1, pOVAOT-1, and gOVAOT-1 mice were analyzed 50–60 d after cell transfer. Histograms are gated on congenic OT-1 T cells; undivided CD44− cells are gated in naive mice, while divided CD44+ cells are gated in pOVA and gOVA mice. Data are pooled from two experiments. NaiveOT-1, n = 4–5; gOVAOT-1, n = 9; pOVAOT-1, n = 10–11. PD-1, *, P = 0.02; LAG-3, *, P = 0.004; CD38, *, P < 0.001. Student’s t test. (D) OT-1 cells in pOVA mice express transcription factors associated with T cell exhaustion. Data are pooled from two experiments. NaiveOT-1, n = 3–7; gOVAOT-1, n = 9–15; pOVAOT-1, n = 11–19. TOX, *, P = 0.02; Eomes, *, P = 0.01. Student’s t test. (E) Coexpression of IRs and transcription factors associated with T cell exhaustion. Data are pooled from two experiments. NaiveOT-1, n = 2–5; gOVAOT-1, n = 4–9; pOVAOT-1, n = 6–10. %PD-1+ Eomes+, *, P = 0.01; %PD-1+ Tox+, *, P < 0.001.