Type 1 diabetes mellitus (T1D) is a chronic autoimmune condition in which the immune system destroys insulin-producing pancreatic β cells. In addition to well-established pathogenic effector T cells, regulatory T cells (Tregs) have also been shown to be defective in T1D. Thus, an increasing number of therapeutic approaches are being developed to target Tregs. However, the role and mechanisms of TGF-β-induced Tregs (iTregs) in T1D remain poorly understood. Here, using a streptozotocin (STZ)-induced preclinical T1D mouse model, we found that iTregs could ameliorate the development of T1D and preserve β cell function. The preventive effect was associated with the inhibition of type 1 cytotoxic T (Tc1) cell function and rebalancing the Treg/Tc1 cell ratio in recipients. Furthermore, we showed that the underlying mechanisms were due to the TGF-β-mediated combinatorial actions of mTOR and TCF1. In addition to the preventive role, the therapeutic effects of iTregs on the established STZ-T1D and nonobese diabetic (NOD) mouse models were tested, which revealed improved β cell function. Our findings therefore provide key new insights into the basic mechanisms involved in the therapeutic role of iTregs in T1D.
This is a preview of subscription content, access via your institution
Subscribe to this journal
Receive 12 digital issues and online access to articles
$119.00 per year
only $9.92 per issue
Rent or buy this article
Get just this article for as long as you need it
Prices may be subject to local taxes which are calculated during checkout
Patterson, C. C. et al. Trends and cyclical variation in the incidence of childhood type 1 diabetes in 26 European centres in the 25 year period 1989-2013: a multicentre prospective registration study. Diabetologia 62, 408–17. (2019).
Livingstone, S. J. et al. Estimated life expectancy in a Scottish cohort with type 1 diabetes, 2008-2010. JAMA 313, 37–44 (2015).
Greenbaum, C. J. et al. Strength in numbers: opportunities for enhancing the development of effective treatments for type 1 diabetes—the TrialNet experience. Diabetes 67, 1216–25. (2018).
Eizirik, D. L., Colli, M. L. & Ortis, F. The role of inflammation in insulitis and beta-cell loss in type 1 diabetes. Nat. Rev. Endocrinol. 5, 219–26. (2009).
Willcox, A., Richardson, S. J., Bone, A. J., Foulis, A. K. & Morgan, N. G. Analysis of islet inflammation in human type 1 diabetes. Clin. Exp. Immunol. 155, 173–81. (2009).
Coppieters, K. T. et al. Demonstration of islet-autoreactive CD8 T cells in insulitic lesions from recent onset and long-term type 1 diabetes patients. J. Exp. Med. 209, 51–60 (2012).
Menart-Houtermans, B. et al. Leukocyte profiles differ between type 1 and type 2 diabetes and are associated with metabolic phenotypes: results from the German Diabetes Study (GDS). Diabetes Care 37, 2326–33. (2014).
Vizler, C. et al. Relative diabetogenic properties of islet-specific Tc1 and Tc2 cells in immunocompetent hosts. J. Immunol. 165, 6314–21. (2000).
Buckner, J. H. Mechanisms of impaired regulation by CD4(+)CD25(+)FOXP3(+) regulatory T cells in human autoimmune diseases. Nat. Rev. Immunol. 10, 849–59. (2010).
Ferraro, A. et al. Expansion of Th17 cells and functional defects in T regulatory cells are key features of the pancreatic lymph nodes in patients with type 1 diabetes. Diabetes 60, 2903–13. (2011).
Ryba-Stanislawowska, M., Skrzypkowska, M., Mysliwiec, M. & Mysliwska, J. Loss of the balance between CD4(+)Foxp3(+) regulatory T cells and CD4(+)IL17A(+) Th17 cells in patients with type 1 diabetes. Hum. Immunol. 74, 701–707 (2013).
Lundsgaard, D., Holm, T. L., Hornum, L. & Markholst, H. In vivo control of diabetogenic T-cells by regulatory CD4+CD25+ T-cells expressing Foxp3. Diabetes 54, 1040–1047 (2005).
Jaeckel, E., von Boehmer, H. & Manns, M. P. Antigen-specific FoxP3-transduced T-cells can control established type 1 diabetes. Diabetes 54, 306–10. (2005).
Marek-Trzonkowska, N. et al. Administration of CD4+CD25highCD127- regulatory T cells preserves beta-cell function in type 1 diabetes in children. Diabetes Care 35, 1817–20. (2012).
Horwitz, D. A., Zheng, S. G. & Gray, J. D. Natural and TGF-beta-induced Foxp3(+)CD4(+) CD25(+) regulatory T cells are not mirror images of each other. Trends Immunol. 29, 429–35. (2008).
Zhou, X. et al. Cutting edge: all-trans retinoic acid sustains the stability and function of natural regulatory T cells in an inflammatory milieu. J. Immunol. 185, 2675–2679 (2010).
Komatsu, N. et al. Pathogenic conversion of Foxp3+ T cells into TH17 cells in autoimmune arthritis. Nat. Med. 20, 62–68 (2014).
Xu, W. et al. Adoptive transfer of induced-Treg cells effectively attenuates murine airway allergic inflammation. Plos One 7, e40314 (2012).
Zheng, S. G., Wang, J. & Horwitz, D. A. Cutting edge: Foxp3+CD4+CD25+ regulatory T cells induced by IL-2 and TGF-beta are resistant to Th17 conversion by IL-6. J. Immunol. 180, 7112–7116 (2008).
Xu, L., Kitani, A., Fuss, I. & Strober, W. Cutting edge: regulatory T cells induce CD4+CD25-Foxp3- T cells or are self-induced to become Th17 cells in the absence of exogenous TGF-beta. J. Immunol. 178, 6725–6729 (2007).
Lan, Q. et al. Induced Foxp3(+) regulatory T cells: a potential new weapon to treat autoimmune and inflammatory diseases? J. Mol. Cell Biol. 4, 22–28 (2012).
Kong, N. et al. Induced T regulatory cells suppress osteoclastogenesis and bone erosion in collagen-induced arthritis better than natural T regulatory cells. Ann. Rheum. Dis. 71, 1567–72. (2012).
Kong, N. et al. Antigen-specific transforming growth factor β-induced Treg cells, but not natural Treg cells, ameliorate autoimmune arthritis in mice by shifting the Th17/Treg cell balance from Th17 predominance to Treg cell predominance. Arthritis Rheum. 64, 2548–58. (2012).
Selvaraj, R. K. & Geiger, T. L. Mitigation of experimental allergic encephalomyelitis by TGF-beta induced Foxp3+ regulatory T lymphocytes through the induction of anergy and infectious tolerance. J. Immunol. 180, 2830–2838 (2008).
Weber, S. E. et al. Adaptive islet-specific regulatory CD4 T cells control autoimmune diabetes and mediate the disappearance of pathogenic Th1 cells in vivo. J. Immunol. 176, 4730–4739 (2006).
Luo, X. et al. Dendritic cells with TGF-beta1 differentiate naive CD4+CD25- T cells into islet-protective Foxp3+ regulatory T cells. Proc. Natl Acad. Sci. USA 104, 2821–2826 (2007).
Jones, C. B. et al. Regulatory T cells control diabetes without compromising acute anti-viral defense. Clin. Immunol. 153, 298–307 (2014).
Elias, D. et al. Autoimmune diabetes induced by the beta-cell toxin STZ. Immunity to the 60-kDa heat shock protein and to insulin. Diabetes 43, 992–998 (1994).
Kantwerk, G., Cobbold, S., Waldmann, H. & Kolb, H. L. 3T. 4 and Lyt-2 T cells are both involved in the generation of low-dose streptozotocin-induced diabetes in mice. Clin. Exp. Immunol. 70, 585–592 (1987).
Nakamura, M., Nagafuchi, S., Yamaguchi, K. & Takaki, R. The role of thymic immunity and insulitis in the development of streptozocin-induced diabetes in mice. Diabetes 33, 894–900 (1984).
Buschard, K. & Rygaard, J. T-lymphocytes transfer streptozotocin induced diabetes mellitus in mice. Acta Pathol. Microbiol Scand. C 86C, 277–82. (1978).
Goyal, S. N. et al. Challenges and issues with streptozotocin-induced diabetes—a clinically relevant animal model to understand the diabetes pathogenesis and evaluate therapeutics. Chem. Biol. Interact. 244, 49–63 (2016).
Shimokawa, C. et al. CD8(+) regulatory T cells are critical in prevention of autoimmune-mediated diabetes. Nat. Commun. 11, 1922 (2020).
Müller, A., Schott-Ohly, P., Dohle, C. & Gleichmann, H. Differential regulation of Th1-type and Th2-type cytokine profiles in pancreatic islets of C57BL/6 and BALB/c mice by multiple low doses of streptozotocin. Immunobiology 205, 35–50 (2002).
Kondo, S. et al. Suppression of insulitis and diabetes in B cell-deficient mice treated with streptozocin: B cells are essential for the TCR clonotype spreading of islet-infiltrating T cells. Int Immunol. 12, 1075–83. (2000).
Cai, W. et al. All trans-retinoic acid protects against acute ischemic stroke by modulating neutrophil functions through STAT1 signaling. J. Neuroinflamm. 16, e175 (2019).
Yadav, M. et al. Neuropilin-1 distinguishes natural and inducible regulatory T cells among regulatory T cell subsets in vivo. J. Exp. Med. 209, 1713–1722 (2012). S1–19.
Zheng, S. G., Wang, J. H., Gray, J. D., Soucier, H. & Horwitz, D. A. Natural and induced CD4+CD25+ cells educate CD4+CD25- cells to develop suppressive activity: the role of IL-2, TGF-beta, and IL-10. J. Immunol. 172, 5213–21. (2004).
Xu, A. et al. TGF-beta-induced regulatory T cells directly suppress B cell responses through a noncytotoxic mechanism. J. Immunol. 196, 3631–41. (2016).
Araki, K. et al. mTOR regulates memory CD8 T-cell differentiation. Nature 460, 108–12. (2009).
Rao, R. R., Li, Q., Odunsi, K. & Shrikant, P. A. The mTOR kinase determines effector versus memory CD8+ T cell fate by regulating the expression of transcription factors T-bet and Eomesodermin. Immunity 32, 67–78 (2010).
Ito, D. et al. mTOR complex signaling through the SEMA4A–Plexin B2 axis is required for optimal activation and differentiation of CD8+T cells. J. Immunol. 195, 934–43. (2015).
Yu, Q., Sharma, A. & Sen, J. M. TCF1 and β-catenin regulate T cell development and function. Immunol. Res 47, 45–55 (2010).
Donnarumma, T. et al. Opposing development of cytotoxic and follicular helper CD4 T cells controlled by the TCF-1-Bcl6 nexus. Cell Rep. 17, 1571–83. (2016).
Danilo, M., Chennupati, V., Silva, J. G., Siegert, S. & Held, W. Suppression of Tcf1 by inflammatory cytokines facilitates effector CD8 T cell differentiation. Cell Rep. 22, 2107–17. (2018).
O’Brien, B. A., Harmon, B. V., Cameron, D. P. & Allan, D. J. Beta-cell apoptosis is responsible for the development of IDDM in the multiple low-dose streptozotocin model. J. Pathol. 178, 176–81. (1996).
Cerwenka, A., Morgan, T. M., Harmsen, A. G. & Dutton, R. W. Migration kinetics and final destination of type 1 and type 2 CD8 effector cells predict protection against pulmonary virus infection. J. Exp. Med. 189, 423–34. (1999).
Morlacchi, S. et al. Regulatory T cells target chemokine secretion by dendritic cells independently of their capacity to regulate T cell proliferation. J. Immunol. 186, 6807–14. (2011).
Dal Secco, V. et al. Tunable chemokine production by antigen presenting dendritic cells in response to changes in regulatory T cell frequency in mouse reactive lymph nodes. Plos One 4, e7696 (2009).
Lin, J. T., Martin, S. L., Xia, L. & Gorham, J. D. TGF-beta 1 uses distinct mechanisms to inhibit IFN-gamma expression in CD4+ T cells at priming and at recall: differential involvement of Stat4 and T-bet. J. Immunol. 174, 5950–5958 (2005).
Green, E. A., Gorelik, L., McGregor, C. M., Tran, E. H. & Flavell, R. A. C. D. 4 CD25+ T regulatory cells control anti-islet CD8+ T cells through TGF-beta-TGF-beta receptor interactions in type 1 diabetes. Proc. Natl Acad. Sci. USA 100, 10878–83. (2003).
Li, L., Ma, Y. & Xu, Y. Follicular regulatory T cells infiltrated the ovarian carcinoma and resulted in CD8 T cell dysfunction dependent on IL-10 pathway. Int. Immunopharmacol. 68, 81–87 (2019).
Shirasaki, T. et al. Impaired interferon signaling in chronic hepatitis C patients with advanced fibrosis via the transforming growth factor beta signaling pathway. Hepatology 60, 1519–30. (2014).
Viel, S. et al. TGF-beta inhibits the activation and functions of NK cells by repressing the mTOR pathway. Sci. Signal. 9, a19 (2016).
Yu, Q. et al. T cell factor 1 initiates the T helper type 2 fate by inducing the transcription factor GATA-3 and repressing interferon-gamma. Nat. Immunol. 10, 992–999 (2009).
Tiemessen, M. M. et al. T Cell factor 1 represses CD8+ effector T cell formation and function. J. Immunol. 193, 5480–5487 (2014).
Das, J. et al. Transforming growth factor beta is dispensable for the molecular orchestration of Th17 cell differentiation. J. Exp. Med. 206, 2407–16. (2009).
Fung, T. H. W., Yang, K. Y. & Lui, K. O. An emerging role of regulatory T-cells in cardiovascular repair and regeneration. Theranostics 10, 8924–38. (2020).
Burzyn, D. et al. A special population of regulatory T cells potentiates muscle repair. Cell 155, 1282–95. (2013).
Dial, C. F., Tune, M. K., Doerschuk, C. M. & Mock, J. R. Foxp3(+) regulatory T cell expression of keratinocyte growth factor enhances lung epithelial proliferation. Am. J. Respir. Cell Mol. Biol. 57, 162–73. (2017).
Dombrowski, Y. et al. Regulatory T cells promote myelin regeneration in the central nervous system. Nat. Neurosci. 20, 674–80. (2017).
Facciabene, A. et al. Tumour hypoxia promotes tolerance and angiogenesis via CCL28 and T(reg) cells. Nature 475, 226–30. (2011).
Ali, N. et al. Regulatory T cells in skin facilitate epithelial stem cell differentiation. Cell 169, 1119–29. (2017).
Sharir, R. et al. Regulatory T cells influence blood flow recovery in experimental hindlimb ischaemia in an IL-10-dependent manner. Cardiovasc. Res. 103, 585–96. (2014).
This work was supported by the National Key R&D Program of China (2017YFA0105803), the general program of the National Natural Science Foundation of China (81770826), the Science and Technology Plan Projects of Guangdong Province (2019B020227003), the Key Special Projects of Medical and Health of Guangzhou City (202007040003), and the 5010 Clinical Research Projects of Sun Yat-sen University (2015015). Y.M.C. designed the experiments and helped write the manuscript. L.Z. and X.M.H. performed most experiments and collected data. P.H.C. and J.L.D. participated in experiments during revision. L.Z., X.M.H., T.L., R.D.P. analyzed the data. Y.L., H.C.L., F.H., G.J.S., and C.C.X. contributed administrative, technical or material support. L.Z. and Y.L. wrote the manuscript. We thank Song Guo Zheng (Professor of Medicine, Department of Internal Medicine, Ohio State University College of Medicine) for sharing his wisdom with us during the course of this research and for providing comments that greatly improved the manuscript. We thank Julie Wang for providing technical support and helping us with the study design.
The authors have no competing interests that might be perceived as influencing the results and/or discussion in this paper. The authors declare no competing interests.
Rights and permissions
Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Zhou, L., He, X., Cai, P. et al. Induced regulatory T cells suppress Tc1 cells through TGF-β signaling to ameliorate STZ-induced type 1 diabetes mellitus. Cell Mol Immunol 18, 698–710 (2021). https://doi.org/10.1038/s41423-020-00623-2
- type 1 diabetes mellitus
- induced regulatory T cells
- type 1 cytotoxic T cells
- mTOR and TCF1
This article is cited by
Ginseng-derived panaxadiol ameliorates STZ-induced type 1 diabetes through inhibiting RORγ/IL-17A axis
Acta Pharmacologica Sinica (2023)
Interactions between islets and regulatory immune cells in health and type 1 diabetes