Abstract
T helper type 1 (TH1) cell development involves interferon-γ (IFN-γ) signaling through signal transducer and activator of transcription 1 (STAT1) and interleukin-12 (IL-12) signaling through STAT4 activation. We examined here T-bet regulation and evaluated the actions of T-bet in STAT1- and STAT4-dependent TH1 development processes. We found that T-bet expression during T cell activation was strongly dependent on IFN-γ signaling and STAT1 activation, but was independent of STAT4. Ectopic T-bet expression strongly increased IFN-γ production in TH2 cells activated by PMA-ionomycin, but weakly increased IFN-γ production in TH2 cells stimulated by IL-12–IL-18 or OVA peptide–antigen-presenting cell stimulation. In contrast, IL-12–IL-18–induced IFN-γ production remained STAT4-dependent despite ectopic T-bet expression. Ectopic T-bet expression selectively induced expression of IL-12Rβ2, but not IL-18Rα, in wild-type and STAT1−/− TH2 cells, but did not extinguish expression of GATA-3 and TH2 cytokines. Finally, ectopic T-bet did not directly induce expression of endogenous T- bet independently of IFN-γ or STAT1. Thus, T-bet is induced by IFN-γ and STAT1 signaling during T cell activation. In addition, T-bet mediates STAT1-dependent processes of TH1 development, including the induction of IL-12Rβ2.
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We thank K. Fredrick for animal husbandry. Supported by grants from the NIH.
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Web Fig. 1.
STAT1 and STAT4 differentially regulate IL-12–induced TH1 development: assessment by ELISA. CD4+ DO11.10 lymph node T cells from wild-type, STAT1-/- or STAT4-/- mice were activated with 0.3 µM OVA peptide in the presence or absence cytokines or anti-cytokine antibodies as indicated for 7 days, collected and restimulated with OVA peptide–APCs without cytokine manipulations for 48 h. Supernatants were collected at 48 h and IL-4 and IFN-γ were measured by ELISA. (GIF 8 kb)
Web Fig. 2.
STAT1 and STAT4 differentially regulate IL-12–induced TH1 development: assessment by RNA blotting. Cells in Web Fig. 1 were lysed on day 7 and total RNA was prepared and analyzed by RNA blot for IL-12Rβ2 and GAPDH expression as described3. (JPG 25 kb)
Web Fig. 3.
Conditional requirement for STAT1 in IL-12Rβ2 expression during TH1 development. Mel-14hi CD4+ cells were sorted from wild-type or STAT1-/- DO11.10 lymph nodes and spleens and activated in the presence of the indicated cytokines and neutralizing anti-cytokine antibodies as described in the Methods. On days 1, 2, 3, 4 and 7 post-activation, cells were collected and stained with isotype control (dashed lines) or anti–IL-12Rβ2 (solid lines). (GIF 28 kb)
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Afkarian, M., Sedy, J., Yang, J. et al. T-bet is a STAT1-induced regulator of IL-12R expression in naïve CD4+ T cells. Nat Immunol 3, 549–557 (2002). https://doi.org/10.1038/ni794
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DOI: https://doi.org/10.1038/ni794
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