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Glucocorticoid-induced tumor necrosis factor receptor–related protein co-stimulation facilitates tumor regression by inducing IL-9–producing helper T cells

Nature Medicine volume 21, pages 1010–1017 (2015)Cite this article

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Abstract

T cell stimulation via glucocorticoid-induced tumor necrosis factor receptor (TNFR)-related protein (GITR) elicits antitumor activity in various tumor models; however, the underlying mechanism of action remains unclear. Here we demonstrate a crucial role for interleukin (IL)-9 in antitumor immunity generated by the GITR agonistic antibody DTA-1. IL-4 receptor knockout (Il4ra−/−) mice, which have reduced expression of IL-9, were resistant to tumor growth inhibition by DTA-1. Notably, neutralization of IL-9 considerably impaired tumor rejection induced by DTA-1. In particular, DTA-1–induced IL-9 promoted tumor-specific cytotoxic T lymphocyte (CTL) responses by enhancing the function of dendritic cells in vivo. Furthermore, GITR signaling enhanced the differentiation of IL-9–producing CD4+ T-helper (TH9) cells in a TNFR-associated factor 6 (TRAF6)- and NF-κB–dependent manner and inhibited the generation of induced regulatory T cells in vitro. Our findings demonstrate that GITR co-stimulation mediates antitumor immunity by promoting TH9 cell differentiation and enhancing CTL responses and thus provide a mechanism of action for GITR agonist–mediated cancer immunotherapies.

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Figure 1: IL-4R signaling is essential for DTA-1–induced tumor growth inhibition.
Figure 2: DTA-1–mediated tumor regression requires IL-9.
Figure 3: GITR-triggered IL-9 production facilitates antitumor CTL responses by enhancing DC function in vivo.
Figure 4: GITR co-stimulation enhances mouse and human TH9 differentiation in vitro.
Figure 5: GITR triggering shifts the balance of iTreg cells to TH9 cells.
Figure 6: TRAF6–NF-κB pathway is associated with GITR-mediated TH9 differentiation.

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Acknowledgements

We thank J.-O. Kim (International Vaccine Institute) for CD45.1 congenic mice, D.H. Chung (Seoul National University College of Medicine) for IL-4–deficient mice, Y.-K. Kim (Ewha Institute of Convergence Medicine) for IL-4Ra–deficient mice, T.H. Watts (University of Toronto) and P.P. Pandolfi (Harvard Medical School) for GITR-deficient mice, C.D. Surh (Pohang University of Science and Technology) and T.A. Chatila (Harvard Medical School) for Foxp3GFP reporter mice, Y. Choi (University of Pennsylvania School of Medicine) for Traf6fl/fl mice, G. Lozano (MD Anderson Cancer Center) for K-Ras transgenic mice, S. Sakaguchi (Osaka University) for the antibody to GITR (DTA-1), J.v. Snick (Ludwig Institute) for the antibody to IL-9 (MM9C1), S.-I. Nishikawa (RIKEN Center for Developmental Biology) for the antibody to c-kit (ACK2), H.-W. Yum (Seoul National University) and D.-H. Kim (Seoul National University) for assistance in setting up a colorectal cancer model, and members of the Kang laboratory for technical support. This work was supported by grants from the Public Welfare & Safety Research Program (20100020843; C.-Y.K.) and the National R&D Program for Cancer Control, Ministry of Health and Welfare (no. 0720500; C.-Y.K.).

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  1. Byung-Seok Kim

    Present address: Current address: Department of Immunology and Center for Inflammation and Cancer, MD Anderson Cancer Center, Houston, Texas, USA (B.-S.K.).,

Authors and Affiliations

  1. Laboratory of Immunology, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, Republic of Korea

    Il-Kyu Kim, Byung-Seok Kim, Choong-Hyun Koh, Kwang-Soo Shin, Ga-Eun Lee, Yeonseok Chung & Chang-Yuil Kang

  2. Department of Molecular Medicine and Biopharmaceutical Science, Graduate School of Convergence Science and Technology, Seoul National University, Seoul, Republic of Korea

    Jae-Won Seok, Jun-Seok Park, Eun-Ah Bae, Hyewon Jeon & Chang-Yuil Kang

  3. Laboratory of Pathophysiology, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, Republic of Korea

    Jaebeom Cho, Yujin Jung & Ho-Young Lee

  4. Academy of Immunology and Microbiology, Institute for Basic Science, Pohang, Republic of Korea

    Daehee Han

  5. Graduate School of Cancer Science and Policy, National Cancer Center, Goyang, Republic of Korea

    Byoung S Kwon

  6. Center for Immunology and Autoimmune Diseases, Institute of Molecular Medicine, University of Texas Medical School at Houston, Houston, Texas, USA

    Yeonseok Chung

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Contributions

I.-K.K., B.-S.K., Y.C. and C.-Y.K. designed the study; I.-K.K., B.-S.K., C.-H.K., J.-W.S., J.-S.P., K.-S.S., E.-A.B., G.-E.L. and H.J. performed experiments; I.-K.K., B.-S.K., C.-H.K., J.-W.S., Y.C. and C.-Y.K. analyzed data; J.C., Y.J., D.H., B.S.K. and H.-Y.L. provided animals and reagents; I.-K.K., Y.C. and C.-Y.K. wrote the manuscript; C.-Y.K. supervised the study.

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Correspondence to Chang-Yuil Kang.

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Kim, IK., Kim, BS., Koh, CH. et al. Glucocorticoid-induced tumor necrosis factor receptor–related protein co-stimulation facilitates tumor regression by inducing IL-9–producing helper T cells. Nat Med 21, 1010–1017 (2015). https://doi.org/10.1038/nm.3922

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