Elsevier

Immunology Letters

Volume 134, Issue 1, 30 November 2010, Pages 7-16
Immunology Letters

1α,25-Dihydroxyvitamin D3 and all-trans retinoic acid synergistically inhibit the differentiation and expansion of Th17 cells

https://doi.org/10.1016/j.imlet.2010.07.002Get rights and content

Abstract

1α,25-Dihydroxyvitamin D3 (1,25D3), the active form of vitamin D3, is an immunoregulatory hormone with beneficial effects on Th1 cell-mediated inflammatory diseases. Although IL-17-producing CD4+ T helper (Th17) cells have been recently identified as novel effector cells, the immunomodulating effects of 1,25D3 on Th17 cells have not been well defined. We confirmed here that 1,25D3 inhibited the generation of Th17 cells in vitro. Interestingly, 1,25D3 synergistically suppressed the generation of Th17 cells by the combination with all-trans retinoic acid (ATRA). 1,25D3 and ATRA suppressed the development of allergen-induced contact hypersensitivity (CHS) in a mouse ear swelling model. In addition, we found that 1,25D3 and ATRA significantly inhibited the development of human Th17 cells from both naïve and memory human CD4+ T cells. 1,25D3 and ATRA effectively suppressed mRNA expressions of IL-1R1, IL-21R, IL-23R, RORC, and AHR in human T cells. ATRA further suppressed IL-6R, whereas 1,25D3 did not. Finally, we found that 1,25D3 and ATRA remarkably blocked IL-22 as well as IL-17 mRNA expression in human memory CD4+ T cells. Thus, we initially reveal that 1,25D3 and ATRA have synergistic effects on the generation of Th17 cells, suggesting that the combination with ATRA would provide a promising novel therapy for Th17 cell-related immune diseases including skin inflammation.

Introduction

It has been well accepted that CD4+ T helper (Th) cells play a pivotal role for the regulation of the immune responses. More than 20 years ago, Mosmann and Coffman [1] defined two distinct CD4+ T cell subsets (Th1 and Th2 cells) in terms of their differential cytokine-producing patterns. Th1 cells, producing IL-2, IFN-γ and TNF-α (type-1 cytokines), have been thought to mediate protective immunity to infectious diseases and tumors [2], [3], [4], [5] by promoting effector functions of macrophages, dendritic cells (DCs), natural killer (NK) cells and cytotoxic T lymphocytes (CTLs). However, it is also well known that abnormal activation of Th1 cells promotes pathogenic inflammatory responses [6] and mediates autoimmune diseases, such as multiple sclerosis and rheumatoid arthritis [7], [8]. On the other hand, Th2 cells, producing IL-4, IL-5, IL-6, IL-10 and IL-13 (type-2 cytokines), have been thought to support the maturation of B cells to plasma cells which produce antibodies, and mediate protective immunity to extracellular pathogens although their excessive activation causes allergic diseases [9], [10]. Therefore, the proper regulation of the Th1/Th2 balance is required for the maintenance of good health.

A seco-steroid hormone, 1α,25-dihydroxyvitamin D3 (1,25D3) is the active form of vitamin D3 and plays a critical role in the maintenance of mineral homeostasis and normal skeletal architecture. Moreover, it acts as a potent biological response modifier including immunomodulatory activities [11]. In relation to the Th1/Th2 balance, 1,25D3 and its derivatives show their immunomodulating activities through their direct effect on naïve CD4+ T cells [12], [13] and their indirect effect on DCs [12], [13], [14]. It has also been demonstrated that the administration of 1,25D3 prevented Th1-mediated autoimmune diseases in animal models [15], [16], [17], [18]. 1,25D3 and its analog also enhance CD4+ CD25+ regulatory T cells in the model of type 1 diabetes [19] and contact hypersensitivity response [20]. In addition, IL-10-producing regulatory T cells can be induced in vitro by 1,25D3 in the presence of dexamethasone [21]. 1,25D3 also represses IL-2 transcriptional activation by direct inhibition of NFAPp/AP-1 complex formation by VDR-retinoid X receptor heterodimer [22]. Despite these immunomodulating effects, VDR ligands have not been widely applied to the therapy for immune diseases, because of the side effect, hypercalcemia. Therefore, less calcemic analogs have been synthesized to overcome the side effect [11]. Another strategy to resolve this is to develop a more efficient protocol to downmodulate the function of pathogenic effector cells using a lower dose of 1,25D3.

Interleukin 17 (IL-17)-producing helper T (Th17) cells have been described as a functionally new CD4+ helper T cell subset [23]. It has been reported that Th17 cells produce IL-21, IL-22, and TNF-α in addition to IL-17A and IL-17F, and have a critical role in host defense against infection by recruiting neutrophils and macrophages to infected tissues. Th17 cells have also been considered as a key player in autoimmune and inflammatory diseases [24], [25], [26], and the high expression of IL-17 has been reported in human diseases including multiple sclerosis [27], [28], rheumatoid arthritis [29], psoriasis [30] and inflammatory bowel disease [31], though some of these diseases had been thought to be mediated by Th1 cells. There are a few studies that demonstrated 1,25D3 suppressed IL-17 production in animal models, such as TNBS colitis [32] and experimental autoimmune uveitis [33], and in human CD4+ T cells [34], [35]. Tang et al. [33] shows that 1,25D3 directly suppresses IL-17 production by CD4+ T cells and also inhibits the production of IL-1, IL-6, TNF-α, and IL-12/IL-23p40 by DCs in mice.

We demonstrated here that 1,25D3 directly inhibits the development and expansion of Th17 cells from both naïve and memory CD4+ T cells. Moreover, we revealed that the combination of 1,25D3 with all-trans retinoic acid (ATRA), a vitamin A metabolite, which was also reported to inhibit Th17 generation and promoted Foxp3+ Treg differentiation [36], [37], exhibited synergistically inhibitory effects on the development of Th17 cells. In the present study, we described the mechanisms of the inhibitory effects by 1,25D3 and ATRA on human Th17 cells, indicating that a combination therapy using 1,25D3 and ATRA would be a novel and beneficial strategy to overcome the hypercalcemic effect of 1,25D3.

Section snippets

Mice

Wild-type BALB/c mice and C57BL/6 mice were purchased from Charles River Breeding Laboratories (Kanagawa, Japan). OVA323–339-specific I-Ad-restricted T cell receptor (TCR) transgenic mice (DO11.10) were kindly donated by Dr. K. Murphy (Washington University School of Medicine, St. Louis, MO, USA). OVA323–339-specific I-Ab-restricted TCR transgenic mice (OT-II) maintained on the C57BL/6-background were kindly provided by F.R. Carbone (University of Melbourne, Victoria, Australia).

1,25D3 inhibits IL-17-producing CD4+ T cells

To address the immunomodulating effects of 1,25D3 on the cytokine profiles of CD4+ helper T cell subsets, CD4+CD62L+ naïve T cells from DO11.10 TCR transgenic mice were cultured under Th0 (IL-2), Th1 (IL-2, IL-12, IFN-γ anti-IL-4), Th2 (IL-2, IL-4, anti-IFN-γ and anti-IL-12) or Th17 (TGF-β, IL-6, anti-IFN-γ, anti-IL-4) conditions in the presence of OVA peptide with or without 1,25D3 for 6 days. As previously reported [12], 1,25D3 slightly suppressed IFN-γ, type-1 cytokine, production and

Discussion

CD4+ helper T cells including Th1, Th2, and Th17 cells are critical for regulation of immune responses, and the disordered activation promotes inflammatory responses and occasionally mediates the pathogenesis in certain autoimmune diseases [7], [8], [24]. Although it has been reported that Vitamin D and its analogs including 1,25D3 have immunomodulating effects on naïve CD4+ T cells to inhibit the development of Th1 cells and promote the development of Th2 cells [12], [13], we demonstrated here

Acknowledgements

We would like to thank Dr. T. Sawada (Shionogi Pharmaceutical Institute Co., Osaka, Japan) and Dr. H. Tashiro (Fujiya Co. Ltd., Hadano, Japan) for their kind donations of rhIL-2 and OVA323–339 peptide, respectively. We also thank Chugai Pharmaceutical Co. Ltd. for its generous donation of Rocaltrol® and rhIL-6. This work was partially supported by a Grant-in-Aid and a National Project “Knowledge Cluster Initiative” (2nd stage, “Sapporo Biocluster Bio-S”), Ministry of Education, Culture, Sports,

References (67)

  • K. Chamoto et al.

    Potentiation of tumor eradication by adoptive immunotherapy with T-cell receptor gene-transduced T-helper type 1 cells

    Cancer Res

    (2004)
  • D. Wakita et al.

    IFN-gamma-dependent type 1 immunity is crucial for immunosurveillance against squamous cell carcinoma in a novel mouse carcinogenesis model

    Carcinogenesis

    (2009)
  • T. Ohkuri et al.

    Identification of novel helper epitopes of MAGE-A4 tumour antigen: useful tool for the propagation of Th1 cells

    Br J Cancer

    (2009)
  • T. Nishimura et al.

    A critical role for antigen-specific Th1 cells in acute liver injury in mice

    J Immunol

    (1999)
  • A.K. Abbas et al.

    Functional diversity of helper T lymphocytes

    Nature

    (1996)
  • A. Hoshino et al.

    STAT6-mediated signaling in Th2-dependent allergic asthma: critical role for the development of eosinophilia, airway hyper-responsiveness and mucus hypersecretion, distinct from its role in Th2 differentiation

    Int Immunol

    (2004)
  • S. Ashino et al.

    CpG-ODN inhibits airway inflammation at effector phase through down-regulation of antigen-specific Th2-cell migration into lung

    Int Immunol

    (2008)
  • S. Nagpal et al.

    Noncalcemic actions of vitamin D receptor ligands

    Endocr Rev

    (2005)
  • A. Boonstra et al.

    1alpha,25-Dihydroxyvitamin d3 has a direct effect on naive CD4(+) T cells to enhance the development of Th2 cells

    J Immunol

    (2001)
  • I. Imazeki et al.

    Immunomodulating effect of vitamin D3 derivatives on type-1 cellular immunity

    Biomed Res

    (2006)
  • J. Matsuzaki et al.

    1alpha,25-Dihydroxyvitamin D3 downmodulates the functional differentiation of Th1 cytokine-conditioned bone marrow-derived dendritic cells beneficial for cytotoxic T lymphocyte generation

    Cancer Sci

    (2006)
  • J.M. Lemire et al.

    1,25-Dihydroxyvitamin D3 prevents the in vivo induction of murine experimental autoimmune encephalomyelitis

    J Clin Invest

    (1991)
  • C. Mathieu et al.

    Prevention of autoimmune diabetes in NOD mice by 1,25 dihydroxyvitamin D3

    Diabetologia

    (1994)
  • J.M. Lemire et al.

    1,25-Dihydroxyvitamin D3 attenuates the expression of experimental murine lupus of MRL/l mice

    Autoimmunity

    (1992)
  • S. Gregori et al.

    A 1alpha,25-dihydroxyvitamin D(3) analog enhances regulatory T-cells and arrests autoimmune diabetes in NOD mice

    Diabetes

    (2002)
  • S. Gorman et al.

    Topically applied 1,25-dihydroxyvitamin D3 enhances the suppressive activity of CD4+CD25+ cells in the draining lymph nodes

    J Immunol

    (2007)
  • F.J. Barrat et al.

    In vitro generation of interleukin 10-producing regulatory CD4(+) T cells is induced by immunosuppressive drugs and inhibited by T helper type 1 (Th1)- and Th2-inducing cytokines

    J Exp Med

    (2002)
  • I. Alroy et al.

    Transcriptional repression of the interleukin-2 gene by vitamin D3: direct inhibition of NFATp/AP-1 complex formation by a nuclear hormone receptor

    Mol Cell Biol

    (1995)
  • L.E. Harrington et al.

    Interleukin 17-producing CD4+ effector T cells develop via a lineage distinct from the T helper type 1 and 2 lineages

    Nat Immunol

    (2005)
  • E. Bettelli et al.

    T(H)-17 cells in the circle of immunity and autoimmunity

    Nat Immunol

    (2007)
  • M. Tajima et al.

    IL-6-dependent spontaneous proliferation is required for the induction of colitogenic IL-17-producing CD8+ T cells

    J Exp Med

    (2008)
  • S. Ashino et al.

    A T(h)17-polarized cell population that has infiltrated the lung requires cells that convert to IFN-{gamma} production in order to induce airway hyperresponsiveness

    Int Immunol

    (2010)
  • C. Lock et al.

    Gene-microarray analysis of multiple sclerosis lesions yields new targets validated in autoimmune encephalomyelitis

    Nat Med

    (2002)
  • Cited by (0)

    View full text