‘Default’ generated neonatal regulatory T cells are hypomethylated at conserved non‐coding sequence 2 and promote long‐term cardiac allograft survival

Regulatory T (Treg) cells play an important role in the maintenance of immune self‐tolerance and homeostasis. We previously reported that neonatal CD4+ T cells have an intrinsic ‘default’ mechanism to become Treg (neoTreg) cells in response to T‐cell receptor (TCR) stimulation. However, the underlying mechanisms are unclear and the effects of neoTreg cells on regulating immune responses remain unknown. Due to their involvement in Foxp3 regulation, we examined the role of DNA methyltransferase 1 (DNMT1) and DNMT3b during the induction of neoTreg cells in the Foxp3gfp mice. The function of neoTreg cells was assessed in an acute allograft rejection model established in RAG2−/− mice with allograft cardiac transplantation and transferred with syngeneic CD4+ effector T cells. Following ex vivo TCR stimulation, the DNMT activity was increased threefold in adult CD4+ T cells, but not significantly increased in neonatal cells. However, adoptively transferred neoTreg cells significantly prolonged cardiac allograft survival (mean survival time 47 days, P < 0·001) and maintained Foxp3 expression similar to natural Treg cells. The neoTreg cells were hypomethylated at the conserved non‐coding DNA sequence 2 locus of Foxp3 compared with adult Treg cells. The DNMT antagonist 5‐aza‐2′‐deoxycytidine (5‐Aza) induced increased Foxp3 expression in mature CD4+ T cells. 5‐Aza‐inducible Treg cells combined with continuous 5‐Aza treatment prolonged graft survival. These results indicate that the ‘default’ pathway of neoTreg cell differentiation is associated with reduced DNMT1 and DNMT3b response to TCR stimulus. The neoTreg cells may be a strategy to alleviate acute allograft rejection.

[1]  T. Sparwasser,et al.  Long-term prevention of chronic allograft rejection by regulatory T-cell immunotherapy involves host Foxp3-expressing T cells. , 2013, Blood.

[2]  Shimon Sakaguchi,et al.  The plasticity and stability of regulatory T cells , 2013, Nature Reviews Immunology.

[3]  Peter Ebert,et al.  T Cell Receptor (TCR) and Transforming Growth Factor β (TGF-β) Signaling Converge on DNA (Cytosine-5)-methyltransferase to Control forkhead box protein 3 (foxp3) Locus Methylation and Inducible Regulatory T Cell Differentiation* , 2013, The Journal of Biological Chemistry.

[4]  U. Beier,et al.  Foxp3+ T-regulatory cells require DNA methyltransferase 1 expression to prevent development of lethal autoimmunity. , 2013, Blood.

[5]  P. Zhang,et al.  GM-CSF contributes to aortic aneurysms resulting from SMAD3 deficiency. , 2013, The Journal of clinical investigation.

[6]  S. Sakaguchi,et al.  Development and maintenance of regulatory T cells. , 2013, Immunity.

[7]  B. Prakken,et al.  A3.14 PD-1 Signalling and Low RORC mRNA Content Influence the TREG/TH17 Balance in Human Cord Blood , 2013 .

[8]  Zachary D. Smith,et al.  DNA methylation: roles in mammalian development , 2013, Nature Reviews Genetics.

[9]  F. Alvarez,et al.  Adoptive transfer of ex vivo expanded regulatory T cells in an autoimmune hepatitis murine model restores peripheral tolerance , 2013, Hepatology.

[10]  C. Daniel,et al.  Therapeutic opportunities for manipulating TReg cells in autoimmunity and cancer , 2012, Nature Reviews Drug Discovery.

[11]  K. Nakai,et al.  T cell receptor stimulation-induced epigenetic changes and Foxp3 expression are independent and complementary events required for Treg cell development. , 2012, Immunity.

[12]  Shane J. Neph,et al.  Foxp3 Exploits a Pre-Existent Enhancer Landscape for Regulatory T Cell Lineage Specification , 2012, Cell.

[13]  P. Vyas,et al.  Azacitidine augments expansion of regulatory T cells after allogeneic stem cell transplantation in patients with acute myeloid leukemia (AML). , 2012, Blood.

[14]  A. Rudensky,et al.  Regulatory T cells: mechanisms of differentiation and function. , 2012, Annual review of immunology.

[15]  J. Lafaille,et al.  Induced CD4+Foxp3+ regulatory T cells in immune tolerance. , 2012, Annual review of immunology.

[16]  S. Reuter,et al.  DC-derived IL-18 drives Treg differentiation, murine Helicobacter pylori-specific immune tolerance, and asthma protection. , 2012, The Journal of clinical investigation.

[17]  Herman Waldmann,et al.  Plasticity of Foxp3(+) T cells reflects promiscuous Foxp3 expression in conventional T cells but not reprogramming of regulatory T cells. , 2012, Immunity.

[18]  Cheong-Hee Chang,et al.  Susceptibility to Acute Mouse Adenovirus Type 1 Respiratory Infection and Establishment of Protective Immunity in Neonatal Mice , 2012, Journal of Virology.

[19]  M. Leal,et al.  Preterm neonates show marked leukopenia and lymphopenia that are associated with increased regulatory T-cell values and diminished IL-7 , 2012, Pediatric Research.

[20]  E. Schiffrin,et al.  T Regulatory Lymphocytes Prevent Aldosterone-Induced Vascular Injury , 2012, Hypertension.

[21]  R. Foà,et al.  Functional analysis and gene expression profile of umbilical cord blood regulatory T cells , 2012, Annals of Hematology.

[22]  T. Jacks,et al.  Expression of tumour-specific antigens underlies cancer immunoediting , 2011, Nature.

[23]  F. Regateiro,et al.  Sustained suppression by Foxp3+ regulatory T cells is vital for infectious transplantation tolerance , 2011, The Journal of experimental medicine.

[24]  Sebastian Reuter,et al.  Helicobacter pylori infection prevents allergic asthma in mouse models through the induction of regulatory T cells. , 2011, The Journal of clinical investigation.

[25]  Jincun Zhao,et al.  IFN-γ– and IL-10–expressing virus epitope-specific Foxp3+ T reg cells in the central nervous system during encephalomyelitis , 2011, The Journal of experimental medicine.

[26]  P. Ye,et al.  Treatment With Interleukin-12/23p40 Antibody Attenuates Acute Cardiac Allograft Rejection , 2011, Transplantation.

[27]  Christophe Benoist,et al.  Stability of the Regulatory T Cell Lineage in Vivo , 2010, Science.

[28]  Zhiyong Guo,et al.  “Default” Generation of Neonatal Regulatory T Cells , 2010, The Journal of Immunology.

[29]  A. Rudensky,et al.  Role of conserved non-coding DNA elements in the Foxp3 gene in regulatory T-cell fate , 2010, Nature.

[30]  Chih-Yi Chen,et al.  The tobacco-specific carcinogen NNK induces DNA methyltransferase 1 accumulation and tumor suppressor gene hypermethylation in mice and lung cancer patients. , 2010, The Journal of clinical investigation.

[31]  Sascha Karberg.  Switching on Epigenetic Therapy , 2009, Cell.

[32]  Peter A. Jones,et al.  Rethinking how DNA methylation patterns are maintained , 2009, Nature Reviews Genetics.

[33]  Annaïg Lan,et al.  The key role of segmented filamentous bacteria in the coordinated maturation of gut helper T cell responses. , 2009, Immunity.

[34]  S. Mazmanian,et al.  The gut microbiota shapes intestinal immune responses during health and disease , 2009, Nature Reviews Immunology.

[35]  Yuka Kanno,et al.  Global mapping of H3K4me3 and H3K27me3 reveals specificity and plasticity in lineage fate determination of differentiating CD4+ T cells. , 2009, Immunity.

[36]  J. Issa,et al.  Mechanisms of resistance to 5-aza-2'-deoxycytidine in human cancer cell lines. , 2009, Blood.

[37]  D. Levy,et al.  Epigenetic Regulation of Foxp3 Expression in Regulatory T Cells by DNA Methylation1 , 2009, The Journal of Immunology.

[38]  K. Kretschmer,et al.  DNA methylation controls Foxp3 gene expression , 2008, European journal of immunology.

[39]  T. Nomura,et al.  Regulatory T Cells and Immune Tolerance , 2008, Cell.

[40]  Keith D. Robertson,et al.  DNA Methylation Inhibitor 5-Aza-2′-Deoxycytidine Induces Reversible Genome-Wide DNA Damage That Is Distinctly Influenced by DNA Methyltransferases 1 and 3B , 2007, Molecular and Cellular Biology.

[41]  W. Leonard,et al.  CREB/ATF-dependent T cell receptor–induced FoxP3 gene expression: a role for DNA methylation , 2007, The Journal of experimental medicine.

[42]  J. Leventhal,et al.  Targeting Acute Allograft Rejection by Immunotherapy With Ex Vivo-Expanded Natural CD4+CD25+ Regulatory T Cells , 2006, Transplantation.

[43]  A. Naji,et al.  Thymic selection of CD4+CD25+ regulatory T cells induced by an agonist self-peptide , 2001, Nature Immunology.

[44]  A. B. Lyons,et al.  Divided we stand: Tracking cell proliferation with carboxyfluorescein diacetate succinimidyl ester , 1999, Immunology and cell biology.

[45]  Luqman,et al.  Bolus injection of aqueous antigen leads to a high density of T‐cell‐receptor ligand in the spleen, transient T‐cell activation and anergy induction , 1998, Immunology.

[46]  Sharon K. Ahluwalia,et al.  Peripheral Education of the Immune System by Colonic Commensal Microbiota , 2012 .

[47]  O. Joffre,et al.  Prevention of acute and chronic allograft rejection with CD4+CD25+Foxp3+ regulatory T lymphocytes , 2008, Nature Medicine.

[48]  H. Ochs,et al.  The immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome (IPEX) is caused by mutations of FOXP3 , 2001, Nature Genetics.