Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Stat4-dependent, T-bet-independent regulation of IL-10 in NK cells

Genes & Immunity volume 9, pages 316–327 (2008)Cite this article

Abstract

Interleukin-10 (IL-10) is intensely studied, yet little is known about the mechanisms that control IL-10 expression. We identified striking similarities between IL-10 and interferon-γ (IFN-γ) regulation in mouse natural killer (NK) cells. Like IFN-γ, IL-10 expression is induced by IL-2 and IL-12 and IL-2+IL-12 stimulation is synergistic. Unlike IFN-γ, neither IL-18 nor Ly-49D cross-linking induced IL-10 expression however. Additionally, the IL-12 homologs IL-23 and IL-27 also do not regulate NK cell-specific IL-10. We determined that a small population of NK cells accounts for IL-10 production. The induction of IL-10 by IL-2+IL-12 treatment in NK cells appears to be biphasic, with an initial burst of expression which diminishes by 12 h but spikes again at 18 h. We determined that much like IFN-γ, Stat4 is largely required for IL-12-induced IL-10. Conversely, we observed normal induction of IL-10 in T-bet-deficient NK cells. We identified a Stat4-binding element in the fourth intron of the Il10 gene, which is completely conserved between mouse and human. This intronic Stat4 motif is within a conserved noncoding sequence, which is also a target for cytokine-induced histone acetylation. These findings highlight tissue- and receptor-specific IL-10 regulatory mechanisms, which may be part of an early feedback loop.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  1. McCulloch CA, Downey GP, El-Gabalawy H . Signalling platforms that modulate the inflammatory response: new targets for drug development. Nat Rev Drug Discov 2006; 5: 864.

    Article  CAS  PubMed  Google Scholar 

  2. Mocellin S, Marincola F, Rossi CR, Nitti D, Lise M . The multifaceted relationship between IL-10 and adaptive immunity: putting together the pieces of a puzzle. Cytokine Growth Factor Rev 2004; 15: 61.

    Article  CAS  PubMed  Google Scholar 

  3. Moore KW, O’Garra A, De Waal MR, Vieira P, Mosmann TR . Interleukin-10. Annu Rev Immunol 1993; 11: 165.

    Article  CAS  PubMed  Google Scholar 

  4. Mege JL, Meghari S, Honstettre A, Capo C, Raoult D . The two faces of interleukin 10 in human infectious diseases. Lancet Infect Dis 2006; 6: 557.

    Article  CAS  PubMed  Google Scholar 

  5. Pestka S, Krause CD, Sarkar D, Walter MR, Shi Y, Fisher PB . Interleukin-10 and related cytokines and receptors. Annu Rev Immunol 2004; 22: 929.

    Article  CAS  PubMed  Google Scholar 

  6. D’Andrea A, Aste-Amezaga M, Valiante NM, Ma X, Kubin M, Trinchieri G . Interleukin 10 (IL-10) inhibits human lymphocyte interferon gamma-production by suppressing natural killer cell stimulatory factor/IL-12 synthesis in accessory cells. J Exp Med 1993; 178: 1041.

    Article  PubMed  Google Scholar 

  7. Fiorentino DF, Zlotnik A, Mosmann TR, Howard M, O’Garra A . IL-10 inhibits cytokine production by activated macrophages. J Immunol 1991; 147: 3815.

    CAS  PubMed  Google Scholar 

  8. Mocellin S, Panelli MC, Wang E, Nagorsen D, Marincola FM . The dual role of IL-10. Trends Immunol 2003; 24: 36.

    Article  CAS  PubMed  Google Scholar 

  9. Moore KW, De Waal MR, Coffman RL, O’Garra A . Interleukin-10 and the interleukin-10 receptor. Annu Rev Immunol 2001; 19: 683.

    Article  CAS  PubMed  Google Scholar 

  10. Mocellin S, Panelli M, Wang E, Rossi CR, Pilati P, Nitti D et al. IL-10 stimulatory effects on human NK cells explored by gene profile analysis. Genes Immun 2004; 5: 621.

    Article  CAS  PubMed  Google Scholar 

  11. Boulland ML, Meignin V, Leroy-Viard K, Copie-Bergman C, Briere J, Touitou R et al. Human interleukin-10 expression in T/natural killer-cell lymphomas: association with anaplastic large cell lymphomas and nasal natural killer-cell lymphomas. Am J Pathol 1998; 153: 1229.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Shibata Y, Foster LA, Kurimoto M, Okamura H, Nakamura RM, Kawajiri K et al. Immunoregulatory roles of IL-10 in innate immunity: IL-10 inhibits macrophage production of IFN-gamma-inducing factors but enhances NK cell production of IFN-gamma. J Immunol 1998; 161: 4283.

    CAS  PubMed  Google Scholar 

  13. Cai G, Kastelein RA, Hunter CA . IL-10 enhances NK cell proliferation, cytotoxicity and production of IFN-gamma when combined with IL-18. Eur J Immunol 1999; 29: 2658.

    Article  CAS  PubMed  Google Scholar 

  14. Chakir H, Lemay AM, Webb JR . Cytokine expression by murine DX5+ cells in response to IL-12, IL-18, or the combination of IL-12 and IL-18. Cell Immunol 2001; 212: 71.

    Article  CAS  PubMed  Google Scholar 

  15. Wakkach A, Fournier N, Brun V, Breittmayer JP, Cottrez F, Groux H . Characterization of dendritic cells that induce tolerance and T regulatory 1 cell differentiation in vivo. Immunity 2003; 18: 605.

    Article  CAS  PubMed  Google Scholar 

  16. Biron CA, Byron KS, Sullivan JL . Severe herpesvirus infections in an adolescent without natural killer cells. N Engl J Med 1989; 320: 1731.

    Article  CAS  PubMed  Google Scholar 

  17. Arase H, Mocarski ES, Campbell AE, Hill AB, Lanier LL . Direct recognition of cytomegalovirus by activating and inhibitory NK cell receptors. Science 2002; 296: 1323.

    Article  CAS  PubMed  Google Scholar 

  18. Cousens LP, Orange JS, Biron CA . Endogenous IL-2 contributes to T cell expansion and IFN-gamma production during lymphocytic choriomeningitis virus infection. J Immunol 1995; 155: 5690.

    CAS  PubMed  Google Scholar 

  19. Bream JH, Curiel RE, Yu CR, Egwuagu CE, Grusby MJ, Aune TM et al. IL-4 synergistically enhances both IL-2- and IL-12-induced IFN-gamma expression in murine NK cells. Blood 2003; 102: 207.

    Article  CAS  PubMed  Google Scholar 

  20. Wolk K, Kunz S, Asadullah K, Sabat R . Cutting edge: immune cells as sources and targets of the IL-10 family members? J Immunol 2002; 168: 5397.

    Article  CAS  PubMed  Google Scholar 

  21. Portielje JE, Lamers CH, Kruit WH, Sparreboom A, Bolhuis RL, Stoter G et al. Repeated administrations of interleukin (IL)-12 are associated with persistently elevated plasma levels of IL-10 and declining IFN-gamma, tumor necrosis factor-alpha, IL-6, and IL-8 responses. Clin Cancer Res 2003; 9: 76.

    CAS  PubMed  Google Scholar 

  22. Trinchieri G . Regulatory role of T cells producing both interferon gamma and interleukin 10 in persistent infection. J Exp Med 2001; 194: F53–F57.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Chan SH, Perussia B, Gupta JW, Kobayashi M, Pospisil M, Young HA et al. Induction of interferon gamma production by natural killer cell stimulatory factor: characterization of the responder cells and synergy with other inducers. J Exp Med 1991; 173: 869.

    Article  CAS  PubMed  Google Scholar 

  24. Bream JH, Hodge DL, Gonsky R, Spolski R, Leonard WJ, Krebs S et al. A distal region in the interferon-gamma gene is a site of epigenetic remodeling and transcriptional regulation by interleukin-2. J Biol Chem 2004; 279: 41249.

    Article  CAS  PubMed  Google Scholar 

  25. Loza MJ, Perussia B . The IL-12 signature: NK cell terminal CD56+high stage and effector functions. J Immunol 2004; 172: 88.

    Article  CAS  PubMed  Google Scholar 

  26. Chan SH, Kobayashi M, Santoli D, Perussia B, Trinchieri G . Mechanisms of IFN-gamma induction by natural killer cell stimulatory factor (NKSF/IL-12). Role of transcription and mRNA stability in the synergistic interaction between NKSF and IL-2. J Immunol 1992; 148: 92.

    CAS  PubMed  Google Scholar 

  27. Rincon M, Enslen H, Raingeaud J, Recht M, Zapton T, Su MS et al. Interferon-gamma expression by Th1 effector T cells mediated by the p38 MAP kinase signaling pathway. EMBO J 1998; 17: 2817.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Morinobu A, Gadina M, Strober W, Visconti R, Fornace A, Montagna C et al. STAT4 serine phosphorylation is critical for IL-12-induced IFN-gamma production but not for cell proliferation. Proc Natl Acad Sci USA 2002; 99: 12281.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Kaplan MH, Sun YL, Hoey T, Grusby MJ . Impaired IL-12 responses and enhanced development of Th2 cells in Stat4-deficient mice. Nature 1996; 382: 174.

    Article  CAS  PubMed  Google Scholar 

  30. Watford WT, Hissong BD, Bream JH, Kanno Y, Muul L, O’Shea JJ . Signaling by IL-12 and IL-23 and the immunoregulatory roles of STAT4. Immunol Rev 2004; 202: 139.

    Article  CAS  PubMed  Google Scholar 

  31. Townsend MJ, Weinmann AS, Matsuda JL, Salomon R, Farnham PJ, Biron CA et al. T-bet regulates the terminal maturation and homeostasis of NK and Valpha14i NKT cells. Immunity 2004; 20: 477.

    Article  CAS  PubMed  Google Scholar 

  32. Im SH, Hueber A, Monticelli S, Kang KH, Rao A . Chromatin-level regulation of the IL10 gene in T cells. J Biol Chem 2004; 279: 46818.

    Article  CAS  PubMed  Google Scholar 

  33. Jones EA, Flavell RA . Distal enhancer elements transcribe intergenic RNA in the IL-10 family gene cluster. J Immunol 2005; 175: 7437.

    Article  CAS  PubMed  Google Scholar 

  34. Young HA, Bream JH . IFN-gamma: recent advances in understanding regulation of expression, biological functions, and clinical applications. Curr Top Microbiol Immunol 2007; 316: 97.

    CAS  PubMed  Google Scholar 

  35. Mehrotra PT, Donnelly RP, Wong S, Kanegane H, Geremew A, Mostowski HS et al. Production of IL-10 by human natural killer cells stimulated with IL-2 and/or IL-12. J Immunol 1998; 160: 2637.

    CAS  PubMed  Google Scholar 

  36. Jeannin P, Delneste Y, Seveso M, Life P, Bonnefoy JY . IL-12 synergizes with IL-2 and other stimuli in inducing IL-10 production by human T cells. J Immunol 1996; 156: 3159.

    CAS  PubMed  Google Scholar 

  37. Gerosa F, Paganin C, Peritt D, Paiola F, Scupoli MT, ste-Amezaga M et al. Interleukin-12 primes human CD4 and CD8T cell clones for high production of both interferon-gamma and interleukin-10. J Exp Med 1996; 183: 2559.

    Article  CAS  PubMed  Google Scholar 

  38. Windhagen A, Anderson DE, Carrizosa A, Williams RE, Hafler DA . IL-12 induces human T cells secreting IL-10 with IFN-gamma. J Immunol 1996; 157: 1127.

    CAS  PubMed  Google Scholar 

  39. Pohl-Koppe A, Balashov KE, Steere AC, Logigian EL, Hafler DA . Identification of a T cell subset capable of both IFN-gamma and IL-10 secretion in patients with chronic Borrelia burgdorferi infection. J Immunol 1998; 160: 1804.

    CAS  PubMed  Google Scholar 

  40. Gerosa F, Nisii C, Righetti S, Micciolo R, Marchesini M, Cazzadori A et al. CD4(+) T cell clones producing both interferon-gamma and interleukin-10 predominate in bronchoalveolar lavages of active pulmonary tuberculosis patients. Clin Immunol 1999; 92: 224.

    Article  CAS  PubMed  Google Scholar 

  41. Jankovic D, Kullberg MC, Feng CG, Goldszmid RS, Collazo CM, Wilson M et al. Conventional T-bet(+)Foxp3(−) Th1 cells are the major source of host-protective regulatory IL-10 during intracellular protozoan infection. J Exp Med 2007; 204: 273.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Anderson CF, Oukka M, Kuchroo VJ, Sacks D . CD4(+)CD25(−)Foxp3(−) Th1 cells are the source of IL-10-mediated immune suppression in chronic cutaneous leishmaniasis. J Exp Med 2007; 204: 285.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Stumhofer JS, Silver JS, Laurence A, Porrett PM, Harris TH, Turka LA et al. Interleukins 27 and 6 induce STAT3-mediated T cell production of interleukin 10. Nat Immunol 2007; 12: 1363.

    Article  Google Scholar 

  44. Vanden ES, Goriely S, De WD, Willems F, Goldman M . IL-23 up-regulates IL-10 and induces IL-17 synthesis by polyclonally activated naive T cells in human. Eur J Immunol 2005; 35: 469.

    Article  Google Scholar 

  45. Dong C, Davis RJ, Flavell RA . MAP kinases in the immune response. Annu Rev Immunol 2002; 20: 55.

    Article  CAS  PubMed  Google Scholar 

  46. Murphy KM, Reiner SL . The lineage decisions of helper T cells. Nat Rev Immunol 2002; 2: 933.

    Article  CAS  PubMed  Google Scholar 

  47. Agnello D, Lankford CS, Bream J, Morinobu A, Gadina M, O’Shea JJ et al. Cytokines and transcription factors that regulate T helper cell differentiation: new players and new insights. J Clin Immunol 2003; 23: 147.

    Article  CAS  PubMed  Google Scholar 

  48. Ma W, Lim W, Gee K, Aucoin S, Nandan D, Kozlowski M et al. The p38 mitogen-activated kinase pathway regulates the human interleukin-10 promoter via the activation of Sp1 transcription factor in lipopolysaccharide-stimulated human macrophages. J Biol Chem 2001; 276: 13664.

    Article  CAS  PubMed  Google Scholar 

  49. Lucas M, Zhang X, Prasanna V, Mosser DM . ERK activation following macrophage FcgammaR ligation leads to chromatin modifications at the IL-10 locus. J Immunol 2005; 175: 469.

    Article  CAS  PubMed  Google Scholar 

  50. Vega MI, Huerta-Yepaz S, Garban H, Jazirehi A, Emmanouilides C, Bonavida B . Rituximab inhibits p38 MAPK activity in 2F7 B NHL and decreases IL-10 transcription: pivotal role of p38 MAPK in drug resistance. Oncogene 2004; 23: 3530.

    Article  CAS  PubMed  Google Scholar 

  51. Zhang S, Kaplan MH . The p38 mitogen-activated protein kinase is required for IL-12-induced IFN-gamma expression. J Immunol 2000; 165: 1374.

    Article  CAS  PubMed  Google Scholar 

  52. Nguyen KB, Watford WT, Salomon R, Hofmann SR, Pien GC, Morinobu A et al. Critical role for STAT4 activation by type 1 interferons in the interferon-gamma response to viral infection. Science 2002; 297: 2063.

    Article  CAS  PubMed  Google Scholar 

  53. Sullivan BM, Jobe O, Lazarevic V, Vasquez K, Bronson R, Glimcher LH et al. Increased susceptibility of mice lacking T-bet to infection with Mycobacterium tuberculosis correlates with increased IL-10 and decreased IFN-gamma production. J Immunol 2005; 175: 4593.

    Article  CAS  PubMed  Google Scholar 

  54. Wang ZY, Sato H, Kusam S, Sehra S, Toney LM, Dent AL . Regulation of IL-10 gene expression in Th2 cells by Jun proteins. J Immunol 2005; 174: 2098.

    Article  CAS  PubMed  Google Scholar 

  55. Saraiva M, Christensen JR, Tsytsykova AV, Goldfeld AE, Ley SC, Kioussis D et al. Identification of a macrophage-specific chromatin signature in the IL-10 locus. J Immunol 2005; 175: 1041.

    Article  CAS  PubMed  Google Scholar 

  56. Zhang X, Edwards JP, Mosser DM . Dynamic and Transient Remodeling of the Macrophage IL-10 Promoter during Transcription. J Immunol 2006; 177: 1282.

    Article  CAS  PubMed  Google Scholar 

  57. Brenner S, Prosch S, Schenke-Layland K, Riese U, Gausmann U, Platzer C . cAMP-induced Interleukin-10 promoter activation depends on CCAAT/enhancer-binding protein expression and monocytic differentiation. J Biol Chem 2003; 278: 5597.

    Article  CAS  PubMed  Google Scholar 

  58. Lin SC . Identification of an NF-Y/HMG-I(Y)-binding site in the human IL-10 promoter. Mol Immunol 2006; 43: 1325.

    Article  CAS  PubMed  Google Scholar 

  59. Cao S, Liu J, Song L, Ma X . The protooncogene c-Maf is an essential transcription factor for IL-10 gene expression in macrophages. J Immunol 2005; 174: 3484.

    Article  CAS  PubMed  Google Scholar 

  60. Shoemaker J, Saraiva M, O’Garra A . GATA-3 directly remodels the IL-10 locus independently of IL-4 in CD4+ T cells. J Immunol 2006; 176: 3470.

    Article  CAS  PubMed  Google Scholar 

  61. Mason LH, Willette-Brown J, Mason AT, McVicar D, Ortaldo JR . Interaction of Ly-49D+ NK cells with H-2Dd target cells leads to Dap-12 phosphorylation and IFN-gamma secretion. J Immunol 2000; 164: 603.

    Article  CAS  PubMed  Google Scholar 

  62. Mason L, Giardina SL, Hecht T, Ortaldo J, Mathieson BJ . LGL-1: a non-polymorphic antigen expressed on a major population of mouse natural killer cells. J Immunol 1988; 140: 4403.

    CAS  PubMed  Google Scholar 

  63. Kalina U, Kauschat D, Koyama N, Nuernberger H, Ballas K, Koschmieder S et al. IL-18 activates STAT3 in the natural killer cell line 92, augments cytotoxic activity, and mediates IFN-gamma production by the stress kinase p38 and by the extracellular regulated kinases p44erk-1 and p42erk-21. J Immunol 2000; 165: 1307.

    Article  CAS  PubMed  Google Scholar 

  64. Bream JH, Young HA, Rice N, Martin MP, Smith MW, Carrington M et al. CCR5 Promoter Alleles and Specific DNA Binding Factors. Science 1999; 284: 223a.

    Article  Google Scholar 

  65. Bream JH, Ping A, Zhang X, Winkler C, Young HA . A single nucleotide polymorphism in the proximal IFN-gamma promoter alters control of gene transcription. Genes Immun 2002; 3: 165.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Dr John J O’Shea for his valuable support of this work. In addition, we thank the Johns Hopkins Becton Dickinson Immune Function Laboratory and Paul Fallon for assistance with flow cytometry-related experiments. We also thank Debbie Hodge and Howard Young for helpful discussions and critical review of this manuscript and for providing Tbet−/− and Ifng−/− mice. This work was supported by National Institutes of Health Grant AI070594 (to JHB).

Author information

Authors and Affiliations

  1. W Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins University, Bloomberg School of Public Health, Baltimore, MD, USA

    L R Grant, C M Hedrich, F Wang, A Moorthy, D Ranatunga & J H Bream

  2. Lymphocyte and Cell Biology Section, Molecular Immunology and Inflammation Branch, National Institute of Musculoskeletal and Skin Diseases, Bethesda, MD, USA

    Z-J Yao & K Wilson

Authors
  1. L R Grant
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Z-J Yao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C M Hedrich
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. F Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A Moorthy
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. K Wilson
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. D Ranatunga
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. J H Bream
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to J H Bream.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Grant, L., Yao, ZJ., Hedrich, C. et al. Stat4-dependent, T-bet-independent regulation of IL-10 in NK cells. Genes Immun 9, 316–327 (2008). https://doi.org/10.1038/gene.2008.20

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/gene.2008.20

Keywords

This article is cited by

Search

Advanced search

Quick links