Extended Data Fig. 7: Ifngr1 deletion from T_reg cells does not affect viral load nor markers associated with CD8⁺ T cell exhaustion during chronic infection.

a, b, Foxp3^Cre-YFP, Ifngr1^L/L or Ifngr1^L/LFoxp3^Cre-YFP mice remained uninfected or were infected with Cl. 13. a, Liver histological score from uninfected mice (n = 2) or mice infected with Cl. 13 8 dpi (n = 4, Foxp3^Cre-YFP; n = 3, Ifngr1^L/L; n = 3, Ifngr1^L/LFoxp3^Cre-YFP). Score determined as: 1= Triaditis, rare sinusoidal lymphocytes; 2= Triaditis, prominent sinusoidal lymphocytes; 3= Triaditis, prominent sinusoidal lymphocytes, single cell apoptosis. b, Serum AST activity was determined in uninfected mice (n = 9, Foxp3^Cre-YFP; n = 2, 1 independent experiment, Ifngr1^L/L; n = 8, Ifngr1^L/LFoxp3^Cre-YFP) or mice infected with Cl. 13, 8 dpi (n = 13, Foxp3^Cre-YFP; n = 14, Ifngr1^L/LFoxp3^Cre-YFP) or 16 dpi (n = 10, Foxp3^Cre-YFP; n = 2, 1 independent experiment, Ifngr1^L/L; n = 9, Ifngr1^L/LFoxp3^Cre-YFP). c, Foxp3^Cre-YFP or Ifngr1^L/LFoxp3^Cre-YFP mice were infected with Cl. 13 and the viral load in serum d16 (n = 12, control; n = 11, Ifngr1-deficient; 2 independent experiments) and d30 (n = 11, control; n = 10, Ifngr1-deficient; 2 independent experiments), kidney (n = 7, control; n = 6, Ifngr1-deficient; 1 independent experiment) and liver (n = 5, control; n = 3, Ifngr1-deficient; 1 independent experiment) were determined. d, Foxp3^Cre-YFP or Ifngr1^L/LFoxp3^Cre-YFP mice were infected with Cl. 13 and treated with 200 µg isotype control (n = 3, control; n = 4, Ifngr1-deficient) or anti-PDL1 (n = 5, control; n = 2, Ifngr1-deficient) every 3 d from 26-38 dpi and the viral load in kidney was determined. e, f, Foxp3^Cre-YFP (n = 19) or Ifngr1^L/LFoxp3^Cre-YFP (n = 16) mice were infected with Cl. 13 and IR co-expression (indicated on top) with PD1 (e), and SPICE plots visualizing multiple combinations of IR co-expression (f), by pooled Tetramer⁺ (GP₃₃, GP₂₇₆, NP₃₉₆) splenic CD8⁺ T cells were determined by flow cytometry. g-j, Foxp3^Cre-YFP or Ifngr1^L/LFoxp3^Cre-YFP mice were infected with Cl. 13 and treated with isotype control (n = 7) or anti-PDL1 (n = 6, control; n = 8, Ifngr1-deficient) as in (d). g, h, Expression of T-bet by bulk splenic CD8⁺ T cells (g) and PD1 by pooled Tetramer⁺ (GP₃₃, GP₂₇₆, NP₃₉₆) splenic CD8⁺ T cells (h) were determined by flow cytometry. i, Percentage of pT_ex (Tcf1⁺TIM3^–) and tT_ex (TCF1^–TIM3⁺) splenic CD8⁺ T cells were determined by flow cytometry. j, Percentage of tT_ex splenic GP33⁺CD8⁺ T cells was determined by flow cytometry. a-j, Data are presented as mean values and represent biologically independent mice and 1 (a, d, and as indicated), 2 (c, g-j) or 3 (b, e, f) independent experiments. a-e, g-j, Statistical significance was determined by a two-way ANOVA with multiple comparisons (a, b) or by an unpaired two-tailed Student’s t-test relative to Foxp3^Cre-YFP mice (c, e) or my multiple unpaired Student’s t-tests (d, g-j) (P values indicated when significant); NS, not significant.

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