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Histone deacetylases 1 and 2 restrain CD4+ cytotoxic T lymphocyte differentiation
Teresa Preglej, Patricia Hamminger, Maik Luu, Tanja Bulat, Liisa Andersen, Lisa Göschl, Valentina Stolz, Ramona Rica, Lisa Sandner, Darina Waltenberger, Roland Tschismarov, Thomas Faux, Thorina Boenke, Asta Laiho, Laura L. Elo, Shinya Sakaguchi, Günter Steiner, Thomas Decker, Barbara Bohle, Alexander Visekruna, Christoph Bock, Birgit Strobl, Christian Seiser, Nicole Boucheron, Wilfried Ellmeier
Teresa Preglej, Patricia Hamminger, Maik Luu, Tanja Bulat, Liisa Andersen, Lisa Göschl, Valentina Stolz, Ramona Rica, Lisa Sandner, Darina Waltenberger, Roland Tschismarov, Thomas Faux, Thorina Boenke, Asta Laiho, Laura L. Elo, Shinya Sakaguchi, Günter Steiner, Thomas Decker, Barbara Bohle, Alexander Visekruna, Christoph Bock, Birgit Strobl, Christian Seiser, Nicole Boucheron, Wilfried Ellmeier
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Research Article Immunology

Histone deacetylases 1 and 2 restrain CD4+ cytotoxic T lymphocyte differentiation

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Abstract

Some effector CD4+ T cell subsets display cytotoxic activity, thus breaking the functional dichotomy of CD4+ helper and CD8+ cytotoxic T lymphocytes. However, molecular mechanisms regulating CD4+ cytotoxic T lymphocyte (CD4+ CTL) differentiation are poorly understood. Here we show that levels of histone deacetylases 1 and 2 (HDAC1-HDAC2) are key determinants of CD4+ CTL differentiation. Deletions of both Hdac1 and 1 Hdac2 alleles (HDAC1cKO-HDAC2HET) in CD4+ T cells induced a T helper cytotoxic program that was controlled by IFN-γ–JAK1/2–STAT1 signaling. In vitro, activated HDAC1cKO-HDAC2HET CD4+ T cells acquired cytolytic activity and displayed enrichment of gene signatures characteristic of effector CD8+ T cells and human CD4+ CTLs. In vivo, murine cytomegalovirus–infected HDAC1cKO-HDAC2HET mice displayed a stronger induction of CD4+ CTL features compared with infected WT mice. Finally, murine and human CD4+ T cells treated with short-chain fatty acids, which are commensal-produced metabolites acting as HDAC inhibitors, upregulated CTL genes. Our data demonstrate that HDAC1-HDAC2 restrain CD4+ CTL differentiation. Thus, HDAC1-HDAC2 might be targets for the therapeutic induction of CD4+ CTLs.

Authors

Teresa Preglej, Patricia Hamminger, Maik Luu, Tanja Bulat, Liisa Andersen, Lisa Göschl, Valentina Stolz, Ramona Rica, Lisa Sandner, Darina Waltenberger, Roland Tschismarov, Thomas Faux, Thorina Boenke, Asta Laiho, Laura L. Elo, Shinya Sakaguchi, Günter Steiner, Thomas Decker, Barbara Bohle, Alexander Visekruna, Christoph Bock, Birgit Strobl, Christian Seiser, Nicole Boucheron, Wilfried Ellmeier

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Figure 6

Enhanced induction of CD4+ CTLs in HDAC1cKO-HDAC2HET mice upon MCMV infection.

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Enhanced induction of CD4+ CTLs in HDAC1cKO-HDAC2HET mice upon MCMV infe...
(A) Experimental strategy: WT and HDAC1cKO-HDAC2HET mice were intraperitoneally injected with 5 × 105 PFU/mouse Δm157-MCMV. Eight days after infection the spleen was isolated, and single-cell suspensions were analyzed by flow cytometry or, prior, restimulated with viral m25 peptide for 4 hours. (B) Flow cytometry analysis of splenocytes isolated from MCMV-infected (or PBS-injected controls) WT and HDAC1cKO-HDAC2HET mice showing CD44, CD62L, CD11a, and CD49d expression on TCRβ+CD4+ cells. (C) Contour plots depict IFN-γ and granzyme B expression in WT and HDAC1cKO-HDAC2HET TCRβ+CD4+ T cells (isolated from MCMV-infected or control mice) restimulated with m25 viral peptide. (D) Summary of experiments described in C. Diagrams depict the ratio of the percentages of either IFN-γ+CD4+CD44hi or granzyme B–positive CD4+CD44hiTCRβ+ cells to the percentages of all CD44hi cells within the TCRβ+CD4+ T cell population isolated from MCMV-infected and control WT and HDAC1cKO-HDAC2HET mice. (E) Contour plots show EOMES, T-bet, and CRTAM expression on TCRβ+CD4+CD44+CD62L– splenocytes isolated from WT and HDAC1cKO-HDAC2HET (MCMV-infected or control) mice. (F) Summary of experiments described in C. Diagrams depict the percentages of TCRβ+CD4+CD44+CD62L– splenocytes expressing EOMES, T-bet, and CRTAM. (B, C, and E) Numbers indicate the percentages of cells in the respective quadrants or gates. (D and F) Each symbol indicates 1 mouse. Horizontal bars indicate the mean. *P < 0.05, **P < 0.01, and ***P < 0.001 (1-way ANOVA analysis followed by Tukey’s multiple-comparisons test). Data are representative (B, C, and E) or show a summary (D and F) of at least 16 (B, C, and E) mice that were analyzed in at least 4 (B, E, and F) or 3 (C and D) independent experiments.

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