Fig. 1: Dysfunctional hepatic virus-specific CXCR6+ CD8 T cells characterized by enhanced CREM activity during persistent hepatotropic infection.
From: A liver immune rheostat regulates CD8 T cell immunity in chronic HBV infection
a, Liver bioluminescence in vivo imaging of Ad–CMV–GOL (resolved), Ad–TTR–GOL (persistent) infected or uninfected mice. P values determined by one-way analysis of variance (ANOVA) with Tukey’s multiple comparisons per timepoint (n = 5). b,c, Expression of CXCR6, CX3CR1 and either CD69 (b) or GzmB (c) by antigen-specific CD45.1+ CD8 T cells in liver and spleen at 45 days post infection (d.p.i.). d–f, Quantification of CXCR6 and CX3CR1 (d), CD69 (e) and GzmB (f) expression data from b and c. P values determined by two-way-ANOVA with Tukey’s multiple comparison for adjusted P value (Padj) (n = 8 (d); n = 5 (e); n = 5 (f)). g, Real-time specific cytotoxicity of CD45.1+ CD8 T cells against OVA257–264 peptide-loaded hepatocytes. P values determined by one-way ANOVA with Tukey’s multiple comparison of area under the curve (AUC) for Padj (n ≥ 3). h, Scheme of CD45.1+ CD8 T cell FACSorting for RNA-seq analysis. i, Principal component (PC) analysis of RNA-seq results (n = 3). j, GSEA in liver CD45.1+CXCR6+ CD8 T cells from resolved (left) and persistent (right) infection for a tissue-residency signature and Hobit- and Blimp1-dependent genes (permutation test with Benjamini–Hochberg false discovery rate (FDR)). NES, normalized enrichment score, k, Differentially expressed genes (DEGs) in liver CD45.1+CXCR6+ CD8 T cells during persistent infection or after resolved infection (red, Padj < 1.31 (P < 0.05 Wald test with Benjamini–Hochberg’s correction) and log2-transformed fold change (FC)> 1 or >−1, n = 3). l, Transcription factor network analysis comparing CD45.1+CXCR6+ CD8 T cells in persistent and resolved infection (n = 3). TFBS, transcription factor-binding site. m,n, 4-1BB expression by virus-specific CD45+ CD8 T cells compared to bulk CD45.1− CD8 T cells at 45 d.p.i. (m) and quantification (n). P values determined by two-way ANOVA with Tukey’s multiple comparison for Padj (n = 5). In a–g,m,n, one out of two or more independent experiments shown; NS, not significant, P ≥ 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Data are mean and s.d. FMO, fluorescence minus one; MFI, geometric mean fluorescence intensity; NA, not analysed.
