Extended Data Fig. 3: AHR controls T cell production of CXCL13.
From: Interferon subverts an AHR–JUN axis to promote CXCL13+ T cells in lupus
a, CXCL13 quantification by ELISA from cells in CRISPR screen without TGF-β. Results from 2 independent experiments using different donors. b, Western blot for CBLB in memory CD4+ T cells treated with control or sgCBLB CRISPR guide (left) and ELISA quantification of CXCL13 from indicated cells (n = 4, 2 biological donors each with 2 technical replicates, p = 0.031). c, Western blot for AHR in cells nucleofected with sgAHR and sgCD8a control. d, ELISA quantification of cytokines from memory CD4+ T cells nucleofected with sgAHR or sgCD8 (n = 12 donors). For CXCL13 p = 4.88e-4 and IL-22 p = 4.88e-4. e, CXCL13 quantification by ELISA in supernatants of memory CD4+ T cells nucleofected with sgAHR or sgCD8a in the presence or absence of TGF-β (n = 8). From left to right, p = 0.0078, 0.0078, 0.0078, 0.0156. f,g, Normalized (to DMSO control) ELISA quantification of indicated cytokines in supernatants of memory (f) or naive CD4+ T cells (g) stimulated under indicated conditions (n = 5–7). For AHRinh and TCDD in f, respectively, p = 7.31e-4 and 0.00304 for CXCL13, and p = 0.00159 and 0.0124 for IL-22. For AHRinh and TCDD in g, respectively, p = 0.0679 and 0.00108 for CXCL13, and p = 0.0192 and 0.0157 for IL-22. h, Normalized (to DMSO control) ELISA quantification of indicated cytokines in supernatants of memory CD4+ T cells stimulated with AHR agonist FICZ, AHR inhibitor GNF-351, or DMSO control (n = 3-4). For FICZ and GNF-351, respectively, p = 0.0109 (GNF-351 only) for CXCL13, and p = 0.0084 and 0.0393 for IL-22. i, Effects of AHR CRISPR deletion (left, n = 10) and pharmacological modulation (middle[n = 9] and right[n = 3]) on IFNγ production measured by ELISA. AHR modulators as in g and h were tested. Results shown normalized to DMSO control. j, Flow cytometry quantification of indicated cytokines in memory CD4 + T cells cultured in polarizing conditions as indicated (n = 6). p = 0.0316 for IL-17. k, ELISA data for CXCL13 (left) and IL-22 (right), normalized to control (DMSO) condition, in supernatants of CD4 + T cells stimulated and cultured with indicated factors. Each dot represents a donor (n = 4-5). l, ELISA data for CD8+ T cells stimulated in the presence of TGF-β with indicated AHR modulators, normalized to DMSO condition per donor (n = 6). For AHRinh and TCDD compared to DMSO, respectively, p = 0.0021 and 0.0038 for CXCL13, and p = 0.0103 and 0.0032 for IL-22. m, ELISA measurement for CXCL13 in supernatants of memory CD8+ T cells nucleofected with sgAHR or control CRISPR guide (n = 6, P = 0.0312). n, Expression of ICOS (left) and CD96 (right) by flow cytometry in memory CD8+ T cells stimulated in indicated conditions, normalized to DMSO condition (n = 8). For AHRinh and TCDD, respectively, p = 0.0158 (AHRinh only) for ICOS, and p = 7.09e-3 and 0.0371 for CD96. Data for f, g, h, i, l and n are shown as mean ± S.D. p-values (NS ≥ 0.05, *p < 0.05, **p < 0.01, ***p < 0.001) by ratio paired t-test for b, f-j, l, n, Wilcoxon test in e, m.
