Activating Transcription Factor 3 Is a Positive Regulator of Human IFNG Gene Expression

IL-12 and IL-18 are essential for Th1 differentiation, whereas the role of IFN-α in Th1 development is less understood. In this microarray-based study, we searched for genes that are regulated by IFN-α, IL-12, or the combination of IL-12 plus IL-18 during the early differentiation of human umbilical cord blood CD4+ Th cells. Twenty-six genes were similarly regulated in response to treatment with IL-12, IFN-α, or the combination of IL-12 plus IL-18. These genes could therefore play a role in Th1 lineage decision. Transcription factor activating transcription factor (ATF) 3 was upregulated by these cytokines and selected for further study. Ectopic expression of ATF3 in CD4+ T cells enhanced the production of IFN-γ, the hallmark cytokine of Th1 cells, whereas small interfering RNA knockdown of ATF3 reduced IFN-γ production. Furthermore, ATF3 formed an endogenous complex with JUN in CD4+ T cells induced to Th1. Chromatin immunoprecipitation and luciferase reporter assays showed that both ATF3 and JUN are recruited to and transactivate the IFNG promoter during early Th1 differentiation. Collectively, these data indicate that ATF3 promotes human Th1 differentiation.

[1]  M. Kaplan,et al.  Impaired IL-12 responses and enhanced development of Th2 cells in Stat4-deficient mice , 1996, Nature.

[2]  W. V. van Blitterswijk,et al.  Serum-free medium for generation and propagation of functional human cytotoxic and helper T cell clones. , 1984, Journal of immunological methods.

[3]  Yuka Kanno,et al.  T-bet regulates Th1 responses through essential effects on GATA-3 function rather than on IFNG gene acetylation and transcription , 2006, The Journal of experimental medicine.

[4]  D. Cantrell,et al.  Sustained IL-12 Signaling Is Required for Th1 Development 1 , 2004, The Journal of Immunology.

[5]  R. Lahesmaa,et al.  Signaling lymphocytic activation molecule (SLAM) is differentially expressed in human Th1 and Th2 cells. , 2000, Journal of immunological methods.

[6]  M. Biffi,et al.  References Subscriptions Permissions Email Alerts The Role of Stat4 in Species-Specific Regulation of Th Cell Development by Type I , 2013 .

[7]  T. Nomura,et al.  Control of Regulatory T Cell Development by the Transcription Factor Foxp3 , 2002 .

[8]  Carrie D. Johnson,et al.  ATF3 regulates MCMV infection in mice by modulating IFN-γ expression in natural killer cells , 2008, Proceedings of the National Academy of Sciences.

[9]  L. Kobierski,et al.  Activating transcription factor-3 stimulates 3',5'-cyclic adenosine monophosphate-dependent gene expression. , 1994, Molecular endocrinology.

[10]  A. Rudensky,et al.  Foxp3 programs the development and function of CD4+CD25+ regulatory T cells , 2003, Nature Immunology.

[11]  Tsonwin Hai,et al.  ATF3 and ATF3 delta Zip. Transcriptional repression versus activation by alternatively spliced isoforms. , 1994, The Journal of biological chemistry.

[12]  M. Biffi,et al.  Selective Expression of an Interleukin-12 Receptor Component by Human T Helper 1 Cells , 1997, The Journal of experimental medicine.

[13]  D. Carvajal,et al.  IL-12-Deficient Mice Are Defective in IFNγ Production and Type 1 Cytokine Responses , 1996 .

[14]  M. Byrne,et al.  Microbial Lipopeptides Induce the Production of IL-17 in Th Cells1 , 2000, The Journal of Immunology.

[15]  R. Johnson,et al.  Identification of a lipopolysaccharide inducible transcription factor in murine macrophages. , 1996, Molecular immunology.

[16]  D. Carvajal,et al.  IL-12-deficient mice are defective in IFN gamma production and type 1 cytokine responses. , 1996, Immunity.

[17]  R. Flavell,et al.  Reprogramming transcription during the differentiation of precursor CD4+ T cells into effector Th1 and Th2 cells. , 1999, Microbes and infection.

[18]  Wolfgang Weninger,et al.  Negative Regulation of TLR-Signaling Pathways by Activating Transcription Factor-31 , 2007, The Journal of Immunology.

[19]  A. D. Panopoulos,et al.  Essential autocrine regulation by IL-21 in the generation of inflammatory T cells , 2007, Nature.

[20]  R. Lahesmaa,et al.  Kinetics and STAT4‐ or STAT6‐mediated regulation of genes involved in lymphocyte polarization to Th1 and Th2 cells , 2003, European journal of immunology.

[21]  R Taub,et al.  Interactions among LRF-1, JunB, c-Jun, and c-Fos define a regulatory program in the G1 phase of liver regeneration , 1992, Molecular and cellular biology.

[22]  J. Farrar,et al.  Distinct Characteristics of Murine STAT4 Activation in Response to IL-12 and IFN-α1 , 2006, The Journal of Immunology.

[23]  G. Crabtree,et al.  Characterization of the nuclear and cytoplasmic components of the lymphoid-specific nuclear factor of activated T cells (NF-AT) complex. , 1993, The Journal of biological chemistry.

[24]  L. Fouser,et al.  Interleukin (IL)-22 and IL-17 are coexpressed by Th17 cells and cooperatively enhance expression of antimicrobial peptides , 2006, The Journal of experimental medicine.

[25]  M. Li,et al.  Activating transcription factor 3 up-regulated by c-Jun NH2-terminal kinase/c-Jun contributes to apoptosis induced by potassium deprivation in cerebellar granule neurons , 2008, Neuroscience.

[26]  R. Lahesmaa,et al.  IL‐12 up‐regulates T‐bet independently of IFN‐γ in human CD4+ T cells , 2005 .

[27]  W. M. Weaver,et al.  The Proximal Regulatory Element of the Interferon-γ Promoter Mediates Selective Expression in T Cells* , 1996, The Journal of Biological Chemistry.

[28]  F. Ramsdell,et al.  An essential role for Scurfin in CD4+CD25+ T regulatory cells , 2003, Nature Immunology.

[29]  R. Lahesmaa,et al.  IL-12 up-regulates T-bet independently of IFN-gamma in human CD4+ T cells. , 2005, European journal of immunology.

[30]  Laurie H Glimcher,et al.  A Novel Transcription Factor, T-bet, Directs Th1 Lineage Commitment , 2000, Cell.

[31]  R. Kastelein,et al.  IGIF Does Not Drive Th1 Development but Synergizes with IL-12 for Interferon-γ Production and Activates IRAK and NFκB , 1997 .

[32]  Matthew J. Stephens,et al.  Global effect of PEG-IFN-alpha and ribavirin on gene expression in PBMC in vitro. , 2004, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[33]  R. Lahesmaa,et al.  Enrichment of nucleofected primary human CD4+ T cells: a novel and efficient method for studying gene function and role in human primary T helper cell differentiation. , 2006, Journal of immunological methods.

[34]  M. Kurimoto,et al.  IFN-alpha and IL-18 synergistically enhance IFN-gamma production in human NK cells: differential regulation of Stat4 activation and IFN-gamma gene expression by IFN-alpha and IL-12. , 2001, European journal of immunology.

[35]  J. Warrington,et al.  Identification and validation of endogenous reference genes for expression profiling of T helper cell differentiation by quantitative real-time RT-PCR. , 2001, Analytical biochemistry.

[36]  M. Kurimoto,et al.  IFN‐α  and IL‐18 synergistically enhance IFN‐γ  production in human NK cells: differential regulation of Stat4 activation and IFN‐γ  gene expression by IFN‐α  and IL‐12 , 2001 .

[37]  B. Samten,et al.  CREB, ATF, and AP-1 Transcription Factors Regulate IFN-γ Secretion by Human T Cells in Response to Mycobacterial Antigen1 , 2008, The Journal of Immunology.

[38]  D. Levy,et al.  IL-6 programs TH-17 cell differentiation by promoting sequential engagement of the IL-21 and IL-23 pathways , 2007, Nature Immunology.

[39]  J. O’Shea,et al.  IL-12 Receptor β2 (IL-12Rβ2)-Deficient Mice Are Defective in IL-12-Mediated Signaling Despite the Presence of High Affinity IL-12 Binding Sites , 2000, The Journal of Immunology.

[40]  R. Kastelein,et al.  IGIF does not drive Th1 development but synergizes with IL-12 for interferon-gamma production and activates IRAK and NFkappaB. , 1997, Immunity.

[41]  I. Gery,et al.  Suppressors of Cytokine Signaling Proteins Are Differentially Expressed in Th1 and Th2 Cells: Implications for Th Cell Lineage Commitment and Maintenance , 2002, The Journal of Immunology.

[42]  R. Rees,et al.  Activation of STAT4 by IL-12 and IFN-alpha: evidence for the involvement of ligand-induced tyrosine and serine phosphorylation. , 1996, Journal of Immunology.

[43]  Terry B. Strom,et al.  IL-21 initiates an alternative pathway to induce proinflammatory TH17 cells , 2007, Nature.

[44]  J. Greenwood,et al.  ATF3 enhances c-Jun-mediated neurite sprouting. , 2003, Brain research. Molecular brain research.

[45]  R. Lahesmaa,et al.  Identification of genes involved in the initiation of human Th1 or Th2 cell commitment , 2005, European journal of immunology.

[46]  W. Henderson,et al.  Activating transcription factor 3 is a negative regulator of allergic pulmonary inflammation , 2008, The Journal of experimental medicine.

[47]  C. Woolf,et al.  ATF3 Increases the Intrinsic Growth State of DRG Neurons to Enhance Peripheral Nerve Regeneration , 2007, The Journal of Neuroscience.

[48]  R. Lahesmaa,et al.  Identification of Novel Genes Regulated by IL-12, IL-4, or TGF-β during the Early Polarization of CD4+ Lymphocytes 1 , 2003, The Journal of Immunology.

[49]  K. Matsushima,et al.  Comprehensive gene expression profile of human activated T(h)1- and T(h)2-polarized cells. , 2001, International immunology.

[50]  Franca Ronchese,et al.  Gene Microarrays Reveal Extensive Differential Gene Expression in Both CD4+ and CD8+ Type 1 and Type 2 T Cells1 , 2001, The Journal of Immunology.

[51]  M. Gadina,et al.  IL-12 receptor beta 2 (IL-12R beta 2)-deficient mice are defective in IL-12-mediated signaling despite the presence of high affinity IL-12 binding sites. , 2000, Journal of immunology.

[52]  M. Yaniv,et al.  The mammalian Jun proteins: redundancy and specificity , 2001, Oncogene.

[53]  M Kurimoto,et al.  Influenza A virus-induced IFN-alpha/beta and IL-18 synergistically enhance IFN-gamma gene expression in human T cells. , 1998, Journal of immunology.

[54]  R. Lahesmaa,et al.  Identification of Novel IL-4/Stat6-Regulated Genes in T Lymphocytes1 , 2003, The Journal of Immunology.

[55]  C. Heusser,et al.  Interferon alpha increases the frequency of interferon gamma-producing human CD4+ T cells , 1993, The Journal of experimental medicine.

[56]  M. Toda,et al.  Immunologic self-tolerance maintained by activated T cells expressing IL-2 receptor alpha-chains (CD25). Breakdown of a single mechanism of self-tolerance causes various autoimmune diseases. , 1995, Journal of immunology.

[57]  IFN-α and IL-12 Induce IL-18 Receptor Gene Expression in Human NK and T Cells1 , 2000, The Journal of Immunology.

[58]  Matthew J. Stephens,et al.  Global effect of PEG-IFN-α and ribavirin on gene expression in PBMC in vitro , 2004 .

[59]  R. Coffman,et al.  Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins. , 1986, Journal of immunology.

[60]  A. Rudensky,et al.  Foxp3 in control of the regulatory T cell lineage , 2007, Nature Immunology.

[61]  Jan Mous,et al.  Touching base , 2000, Nature Genetics.

[62]  T. Aso,et al.  Homocysteine-responsive ATF3 gene expression in human vascular endothelial cells: activation of c-Jun NH(2)-terminal kinase and promoter response element. , 2000, Blood.

[63]  Kathleen A. Kennedy,et al.  Systems biology approaches identify ATF3 as a negative regulator of Toll-like receptor 4 , 2006, Nature.

[64]  R. Flavell,et al.  Reprogramming the signalling requirement for AP-1 (activator protein-1) activation during differentiation of precursor CD4+ T-cells into effector Th1 and Th2 cells. , 1997, Genes and function.

[65]  T. Aune,et al.  Regulation of the activity of IFN-gamma promoter elements during Th cell differentiation. , 1998, Journal of immunology.