Fig. 7: TIGIT controls Th1 and Th17 cell differentiation through acetyl-CoA generation and histone acetylation. | Nature Communications

Fig. 7: TIGIT controls Th1 and Th17 cell differentiation through acetyl-CoA generation and histone acetylation.

From: TIGIT deficiency promotes autoreactive CD4+ T-cell responses through a metabolic‒epigenetic mechanism in autoimmune myositis

Fig. 7

a ATP-citrate lyase (ACLY) controls the conversion of TCA-derived citrate into acetyl-coenzyme A (CoA) for histone acetylation. b–f CD4+ T cells from HC or PM patients were stimulated with αCD3/CD28 in the presence of UK5099 (20 μM), recombinant human CD155-Fc (10 μg/ml), or control IgG1 for 3 d. b Immunoblot analysis of ACLY expression in CD4+ T cells and representative bands of six biologically independent replicates. c Acetyl-CoA levels in whole-cell lysates and the cytosolic and mitochondrial fractions of CD4+ T cells and data from four biologically independent replicates. d Immunoblot analysis of H3K9 and H3K27 acetylation in CD4+ T cells from PM patients or HC. Representative bands of six biologically independent replicates. e, f Immunoblot analysis of ACLY and H3K9 and H3K27 acetylation in CD4+ T cells. Representative bands of four biologically independent replicates. g–i CD4+ T cells from Tigit+/+ or Tigit−/− mice were stimulated with αCD3/CD28. g Immunoblot analysis of ACLY expression in mouse CD4+ T cells. h Acetyl-CoA in whole-cell lysates, cytosolic and mitochondrial fractions of mouse CD4+ T cells. Data from four biologically independent replicates. i Immunoblot analysis of acetylated H3K9 and H3K27 in mouse CD4+ T cells and representative bands. j, k CD4+ T cells from HC or PM were stimulated with αCD3/CD28 in the presence of UK5099, ETO or BPTES for 3 d. Immunoblot analysis of ACLY (j), acetylated H3K9 and H3K27 (k) in CD4+ T cells. l, m Tigit−/− or Tigit+/+ CD4+ T cells were stimulated with αCD3/CD28 in the presence of UK5099, ETO, or BPTES for 3 d. Immunoblot analysis of ACLY (l), acetylated H3K9 and H3K27 (m) in mouse CD4+ T cells treated with UK5099, ETO, or BPTES. n Quantification of H3K9 and H3K27 acetylation in CD4+ T cells from Tigit−/− or Tigit+/+ mice at the Il17a, Il17f, and Ifng promoters and CNS2 and CNS22 by ChIP‒qPCR. o–t Naive CD4+ T cells were cultured under Th1 or Th17 conditions in the presence of C646 (1 μM) or vehicle for 5 d. Flow cytometric analysis of the expression of T-bet, IFNγ (o–q), RORγt, and IL-17A (r–t) in CD4+ T cells. For c, h, p, q, n = four biologically independent replicates. n n = three biologically independent replicates. s, t, n = five biologically independent replicates. e, f, j–m The experiment was repeated three times independently with similar results. Data are mean ± SEM. Statistics were performed using one-way ANOVA followed by multiple comparisons adjustments for c and h, two-tailed unpaired Student’s t test for (n), and two-way ANOVA followed by multiple comparisons adjustments for p, q, s, t.

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