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. 2019 Jun 4;216(8):1791–1808. doi: 10.1084/jem.20190173

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© 2019 Huang et al.

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Figure 3. — IL-27 and IL-27R drive CXCR5⁺ CD8⁺ T cell expansion following IFNAR1 blockade. (A) Percentages and total numbers of GP33-41–specific CXCR5⁺ TIM3⁻ follicular cytotoxic CD8⁺ T cells in WT and congenic IL-12 p35-deficient (Il12a⁻^/⁻), IL-12 p40-deficient (Il12b⁻^/⁻), IL-27 EBI3-deficient (Ebi3⁻^/⁻), and IL-6–deficient (Il6⁻^/⁻ mice treated with IgG1 isotype control or aIFNAR1 antibodies and infected with Cl13. Splenic CXCR5⁺ CD8⁺ T cells were analyzed at 9 dpi. (B) Percentages and total numbers of virus-specific CXCR5⁺ TIM3⁻ follicular cytotoxic CD8⁺ T cells in WT and congenic IL-27R–deficient (Il27ra⁻^/⁻) mice treated with IgG1 isotype control or aIFNAR1 antibodies and infected with Cl13. Splenic CXCR5⁺ CD8⁺ T cells were analyzed at 9 dpi. Bottom: Ratio of individual splenocyte populations in IL-27R–deficient versus WT mice treated with isotype or aIFNAR1. (C) Percentages and total numbers of virus-specific CXCR5⁺ TIM3⁻ CD8⁺ T cells in WT mice that were treated with recombinant IL-27 protein on days 0–8 of Cl13 infection by intraperitoneal injection; data shown for 9 dpi. (D) Dynamics of mRNA expression of IL-27 p28 (Il27) and Stat1 in splenocytes from mice treated with aIFNAR1 or isotype control through 0–7 dpi with Cl13. (E) Expression of GP130 protein on P14 cells from aIFNAR1- or isotype-treated mice infected with Cl13, measured by MFI. (F) Normalized mRNA expression of IL-27R (Il27ra) on virus-specific P14 cells from naive mice or mice treated with isotype or aIFNAR1 and infected with Cl13. Bars represent mean ± SD. (G) Level of IL-27R subunits IL-27RA and GP130 on virus-specific CD8⁺ T cells treated with IFNβ or vehicle. Right: mRNA level of Il6st (encoding GP130) on WT CXCR5⁺ CD8⁺ T cells from Cl13-infected mice treated with vehicle or IFN-I and vehicle-treated CXCR5⁻ CD8⁺ T cells. Numbers on flow plots (A–C) indicate the frequency of each gated population; axes in A–C and horizontal axes in E and G indicate log₁₀ fluorescence. Experiments were performed with three to five mice per group, and data are representative of three independent experiments. Statistical comparisons of experimental groups were performed using Student’s two-tailed t test: not significant (n.s.), P > 0.05; *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.0001. rmIL27, recombinant mouse IL-27 protein; wt, wild type. See also Fig. S3.