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. Author manuscript; available in PMC: 2018 Feb 1.
Published in final edited form as: J Immunol. 2016 Dec 28;198(3):1142–1155. doi: 10.4049/jimmunol.1601297

FIGURE 1.

FIGURE 1

Blimp-1 is required for ThCTL differentiation. (A) WT (circle) or Blimp-1 CKO (square) naïve OT-II CD4 cells were transferred into B6.Thy1.1 mice and infected with A/PR8-OVAII. 8 dpi OT-II effectors were isolated from pooled lungs and assayed for ex vivo cytotoxicity against peptide-pulsed targets (representative of 2 independent experiments, n=5 mice per group each). Significance is shown between WT and Blimp-1 CKO OT-II. (B) CD4 cells isolated from pooled lungs (n = 10–15 mice) from WT or Blimp-1 CKO mice infected with A/PR8 at 8 dpi was assayed for ex-vivo cytotoxicity (pooled data of 2 independent experiments). (C) In vivo killing of peptide-pulsed targets in WT or Blimp-1 CKO mice infected with A/PR8 at 8 dpi (n = 5 mice). Right: Representative flow plot of targets (CFSEloMHC-II+) and bystander cells (CFSEhiMHC-II+). WT or Blimp-1 CKO mice were infected with A/PR8 and the phenotype of lung CD4 cells assayed at 8 dpi for (D) PD-1, (E) CD27, (F) binding to P-selectin (n = 3–5 mice, representative of two independent experiments). (G) Representative histograms of GFP expression of A/PR8-infected Prdm1gfp/+ (open) or Prdm1+/+ (filled) mice at 8 dpi, gated on CD4 from the lung (left), draining lymph node (middle) or spleen (right). (H) Representative flow plot of NKG2A/C/E and Blimp-1 expression of CD4 cells from Blimp-1gfp/+ mice infected with A/PR8 at 8 dpi. (I) WT or Blimp-1 CKO mice were infected with A/PR8 and the percentage of CD4 T cells expressing NKG2A/C/E on 8 dpi was quantified. (J) Same experimental setup as in (A), representative flow plot of NKG2A/C/E expression on WT or Blimp-1 CKO OT-II lung CD4 cells at 8 dpi. Percent (K) NKG2A/C/E+ of OT-II effectors recovered from the lung, dLN, and spleen. (L) Numbers of NKG2A/C/E OT-II effectors recovered from the lung. (J-L n=3–4 mice per group, representative of 2 independent experiments, K= pooled data). Error bars represent SD and significant differences were determined with two-way ANOVA and Fisher’s Least Significant Difference test (A & B) and unpaired two-tailed Student’s t-tests (C–L) where (α = 0.05, * p <0.05, ** p < 0.005, *** p < 0.001, and **** p < 0.0001).