Skip to main content
SpringerLink
Log in

CpG DNA induced IL-12 p40 gene activation is independent of STAT1 activation or production of interferon consensus sequence binding protein

  • Original Paper
  • Published:
Journal of Biomedical Science

Abstract

The host immune system responds to CpG motifs in bacterial DNA by rapidly producing proinflammatory cytokines. The host response to CpG DNA resembles, in many ways, the response to bacterial lipopolysaccharide (LPS). While both agents can induce a powerful inflammatory response, CpG DNA promotes Th1 and suppresses Th2 immunity. The regulation of macrophage IL-12 p40 secretion in response to stimulation with LPS and interferon-γ (IFN-γ) is dependent on the action of a nuclear transacting factor, interferon consensus sequence binding protein (ICSBP), a STAT1-dependent gene product. We found that CpG DNA induced IL-12 p40 secretion by macrophages from mice with either disrupted STAT1 or ICSBP genes. Additionally, CpG DNA did not induce translocation of transcription factors that bind to the gamma-activated sequence in the ICSBP gene nor did CpG DNA induce ICSBP transcription. CpG DNA, which activates macrophages by the TLR9 pathway, is a strong inducer of IL-12 p40, yet does so independently of IFN-γ, STAT1 or ICSBP. Thus, CpG DNA-induced IL-12 p40 secretion is mediated by one or more signaling elements distinct from those induced by either LPS or IFN-γ.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Arditi M, Zhou J, Torres M, Durden DL, Stins M, Kim KS. Lipopolysaccharide stimulates the tyrosine phosphorylation of mitogen-activated protein kinases p44, p42, and p41 in vascular endothelial cells in a soluble CD14-dependent manner. J Immunol 155:3994–4003;1995.

    PubMed  Google Scholar 

  2. Aste-Amezaga M, Ma X, Sartori A, Trinchieri G. Molecular mechanisms of the induction of IL-12 and its inhibition by IL-10. J Immunol 160:5936–5944;1998.

    PubMed  Google Scholar 

  3. Ballas ZK, Rasmussen WL, Krieg AM. Induction of NK activity in murine and human cells by CpG motifs in oligodeoxynucleotides and bacterial DNA. J Immunol 157:1840–1845;1996.

    PubMed  Google Scholar 

  4. Brazolot Millan CL, Weeratna R, Krieg AM, Siegrist CA, Davis HL. CpG DNA can induce strong Th 1 humoral and cell-mediated immune responses against hepatitis B surface antigen in young mice. Proc Natl Acad Sci USA 95:15553–15558;1998.

    Google Scholar 

  5. Chace JH, Hooker NA, Mildenstein KL, Krieg AM, Cowdery JS. Bacterial DNA-induced NK cell IFN-γ production is dependent on macrophage secretion of IL-12. Clin Immunol Immunopathol 84:185–193;1997.

    Article  PubMed  Google Scholar 

  6. Cowdery JS, Boerth NJ, Norian LA, Myung PS, Koretzky GA. Differential regulation of the IL-12 p40 promoter and of p40 secretion by CpG DNA and lipopolysaccharide. J Immunol 162:6770–6775;1999.

    PubMed  Google Scholar 

  7. Cowdery JS, Chace JH, Yi A, Krieg AM. Bacterial DNA induces NK cells to produce IFN-γ in vivo and increases the toxicity of lipopolysaccharides. J Immunol 156:4570–4575;1996.

    PubMed  Google Scholar 

  8. Crowley MT, Costello PS, Fitzer-Attas CJ, Turner M, Meng F, Lowell C, Tybulewicz VLJ, Defranco AL. A critical role for syk in signal transduction and phagocytosis mediated by FCγ receptors on macrophages. J Exp Med 186:1027–1039;1997.

    Article  PubMed  Google Scholar 

  9. Davis HL, Suparto II, Weeratna RR, Jumintarto, Iskandriati DD, Chamzah SS, Ma'ruf AA, Nente CC, Pawitri DD, Krieg AM, Heriyanto, Smits W, Sajuthi DD. CpG DNA overcomes hyporesponsiveness to hepatitis B vaccine in orangutans. Vaccine 18:1920–1924;2000.

    Article  PubMed  Google Scholar 

  10. Giese NA, Gabriele L, Doherty TM, Klinman DM, Tadesse-Heath L, Contursi C, Epstein SL, Morse III HC. Interferon (IFN) consensus sequence-binding protein, a transcription factor of the IFN regulatory factor family regulates immune responses in vivo through control of interleukin 12 expression. J Exp Med 186:1535–1546;1997.

    Article  PubMed  Google Scholar 

  11. Hacker H, Vabulas RM, Takeuchi O, Hoshino K, Akira S, Wagner H. Immune cell activation by bacterial CpG-DNA through myeloid differentiation marker 88 and tumor necrosis factor receptor-associated factor (TRAF)6. J Exp Med 192:595–600;2000.

    PubMed  Google Scholar 

  12. Halpern MD, Kurlander RJ, Pisetsky DS. Bacterial DNA induces murine interferon-γ production by stimulation of interleukin 12 and tumor necrosis factor-γ. Cell Immunol 167:72–78;1996.

    Article  PubMed  Google Scholar 

  13. Hemmi H, Takeuchi O, Kawai T, Kaisho T, Sato S, Sanjo H, Matsumoto M, Hoshino K, Wagner H, Takeda K, Akira S. A toll-like receptor recognizes bacterial DNA. Nature 408:740–745;2000.

    Article  PubMed  Google Scholar 

  14. Hirschfeld M, Ma Y, Weis JH, Vogel SN, Weis JJ. Cutting edge: Repurification of lipopolysaccharide eliminates signaling through both human and murine toll-like receptor 2. J Immunol 165:618–622;2000.

    PubMed  Google Scholar 

  15. Hirschfeld M, Weis JJ, Toshchakov V, Salkowski CA, Cody MJ, Ward DC, Qureshi N, Michalek SM, Vogel SN. Signaling by toll-like receptor 2 and 4 agonists results in different gene expression in murine macrophages. Infect Immun 69:1477–1482;2001.

    Article  PubMed  Google Scholar 

  16. Holtschke T, Lohler J, Kanno Y, Fehr T, Giese N, Rosenbauer F, Lou J, Knobeloch KP, Gabriele L, Waring JF, Bachmann MF, Zinkernagel R, Morse III HC, Ozato K, Horak I. Immunodeficiency and chronic myelogenous leukemia-like syndrome in mice with a targeted mutation of the ICSBP gene. Cell 87:307–317;1996.

    Article  PubMed  Google Scholar 

  17. Kanno Y, Kozak CA, Schindler C, Driggers PH, Ennist DL, Gleason SL, Darnell JE Jr, Ozato K. The genomic structure of the murine ICSBP gene reveals the presence of the gamma interferon-responsive element, to which ISGF3 alpha subunit (or similar) molecule binds. Mol Cell Biol 13:3951–3963;1993.

    PubMed  Google Scholar 

  18. Kantakamalakul W, Politis AD, Marecki S, Sullivan T, Ozato K, Fenton MJ, Vogel SN. Regulation of IFN consensus sequence binding protein expression in murine macrophages. J Immunol 162:7417–7425;1999.

    PubMed  Google Scholar 

  19. Kimbrell DA, Beutler B. The evolution and genetics of innate immunity. Nat Rev Genet 2:256–267;2001.

    Article  PubMed  Google Scholar 

  20. Klinman DM, Yi A, Beaucage SL, Conover J, Krieg AM. CpG motifs present in bacterial DNA rapidly induce lymphocytes to secrete interleukin 6, interleukin 12, and interferon-γ. Proc Natl Acad Sci USA 93:2879–2883;1996.

    Google Scholar 

  21. Krieg AM, Love-Homan L, Yi A, Harty JT. CpG DNA induces sustained IL-12 expression in vivo and resistance toListeria monocytogenes challenge. J Immunol 161:2428–2434;1998.

    PubMed  Google Scholar 

  22. Krieg AM, Yi A, Matson S, Walschmidt TJ, Bishop GA, Teasdale R, Koretzky GA, Klinman DM. CpG motifs in bacterial DNA trigger direct B-cell activation. Nature 374:546–549;1995.

    Article  PubMed  Google Scholar 

  23. Lamont AG, Adorini L. IL-12: A key cytokine in immune regulation. Immunol Today 15:214–217;1996.

    Article  Google Scholar 

  24. Liu HM, Nwebrough SE, Bhatia SK, Krieg AM, Weiner GJ. Immunostimulatory CpG oligodeoxynucleotides enhance the immune response to vaccine strategies involving granulocyte-macrophage colony stimulating factor. Blood 92:3730–3736;1998.

    PubMed  Google Scholar 

  25. Ma X, Gri G, Trinchieri G. A novel ets-2-related nuclear factor is involved in transcriptional activation of the human interleukin-12 p40 gene promoter in response to interferon-γ and LPS stimulation of monocytic cells. Ann NY Acad Sci 795:357–360;1996.

    PubMed  Google Scholar 

  26. Ma X, Neurath M, Gri G, Trinchieri G: Identification and characterization of a novel ets-2-related nuclear complex implicated in the activation of the human interleukin-12 p40 gene promoter. J Biol Chem 272:10389–10395;1997.

    Google Scholar 

  27. Ma X, Chow JM, Gri G, Carra G, Gerosa F, Wolf SF, Dzialo R, Trinchieri G. The interleukin 12 p40 gene promoter is primed by interferon-γ in monocytic cells. J Exp Med 183:147–157;1996.

    PubMed  Google Scholar 

  28. Messina JP, Gilkeson GS, Pisetsky DS. Stimulation of in vitro murine lymphocyte proliferation by bacterial DNA. J Immunol 147:1759–1764;1991.

    PubMed  Google Scholar 

  29. Murphy TL, Cleveland MG, Kulesza P, Magram J, Murphy KM. Regulation of interleukin 12 p40 expression through an NF-κB half-site. Mol Cell Biol 15:5258–5267;1995.

    PubMed  Google Scholar 

  30. Muzio M, Natoli G, Saccani S, Levrero M, Mantovani A. The human toll signaling pathway: Divergence of nuclear factor κB and JNK/SAPK activation upstream of tumor necrosis factor receptor-associated factor 6 (TRAF6). J Exp Med 187:2097–2101;1998.

    Article  PubMed  Google Scholar 

  31. Plevy SE, Gemberling JHM, Hsu S, Dorner AJ, Smale ST: Multiple control elements mediate activation of the murine and human interleukin 12 p40 promoters: Evidence of functional synergy between C/EBP and rel proteins. Mol Cell Biol 17:4572–4588;1997.

    PubMed  Google Scholar 

  32. Politis AD, Ozato K, Coligan JE, Vogel SN: Regulation of IFN-γ-induced nuclear expression of IFN consensus sequence binding protein in murine peritoneal macrophages. J Immunol 152:2270–2278;1994.

    PubMed  Google Scholar 

  33. Poltorak A, He X, Smirnova I, Liu M, VanHuffel C, Du X, Birdwell D, Alejos E, Silva M, Galanos C, Freudenberg M, Ricciardi-Castagnoli P, Layton B, Beutler B. Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: Mutations inTlr4 gene. Science 282:2085–2088;1998.

    Article  PubMed  Google Scholar 

  34. Qureshi ST, Lariviere L, Leveque G, Clermont S, Moore KJ, Gros P, Malo D. Endotoxin-tolerant mice have mutations in Toll-like receptor 4 (Tlr4). J Exp Med 189:615–625;1999.

    Article  PubMed  Google Scholar 

  35. Re F, Strominger JL. Toll-like receptor 2 (TLR2) and TLR4 differentially activate human dendritic cells. J Biol Chem 276:37692–37699;2001.

    Google Scholar 

  36. Roman M, Martin-Orozco E, Goodman JS, Nguyen M, Sato Y, Ronaghy A, Kornbluth RS, Richman DD, Carson DA, Raz E. Immunostimulatory DNA sequences function as T helper-1-promoting adjuvants. Nat Med 3:849–854;1997.

    Article  PubMed  Google Scholar 

  37. Scharton-Keresten T, Contursi C, Masumi A, Sher A, Ozato K. Interferon consensus sequence binding protein-deficient mice display impaired resistance to intracellular infection due to a primary defect in interleukin 12 p40 induction. J Exp Med 186:1523–1534;1997.

    Article  PubMed  Google Scholar 

  38. Sweet MJ, Campbell CC, Sester DP, Xu D, McDonald RC, Stacey KJ, Hume DA, Liew FY. Colony-stimulating factor-1 suppresses responses to CpG DNA and expression of Toll-like receptor 9, but enhances responses to lipopolysaccharide in murine macrophages. J Immunol 168:392–399;2002.

    PubMed  Google Scholar 

  39. Thompson CB, Wang CY, Ho JC, Bohjanen RP, Petyrniak B, Junes SC, Miesfeldt H, Zhang L, Nabel GJ, Karpinski B, Leiden JN.cis-Acting sequences required for inducible interleukin-2 enhancer function find a novel Ets-related protein Elf-1. Mol Cell Biol 12:1043–1053;1992.

    PubMed  Google Scholar 

  40. Trinchieri G. Interleukin-12:A proinflammatory cytokine with immunoregulatory functions that bridge innate resistance and antigen-specific adaptive immunity. Annu Rev Immunol 13:251–276;1995.

    PubMed  Google Scholar 

  41. Ulevitch RJ, Tobias PS. Receptor-dependent mechanisms of cell stimulation by bacterial endotoxin. Annu Rev Immunol 13:437–457;1995.

    Article  PubMed  Google Scholar 

  42. Wang I, Contursi C, Masumi A, Ma X, Trinchieri G, Ozato K. An IFN-γ-inducible transcription factor, IFN consensus sequence binding protein (ICSBP), stimulates IL-12 p40 expression in macrophages. J Immunol 165:271–279;2000.

    PubMed  Google Scholar 

  43. Weiner GJ, Liu HM, Wooldridge JE, Dahle CE, Krieg AM. Immunostimulatory oligodeoxynucleotides containing the CpG motif are effective immune adjuvants in tumor antigen immunization. Proc Natl Acad Sci USA 94:10833–10837;1997.

    Google Scholar 

  44. Wu C, Maeda H, Contursi C, Ozato K, Seder RA. Differential requirement of IFN consensus sequence binding protein for the production of IL-12 and induction of Th1-type cells in response to IFN-γ. J Immunol 162:807–812;1999.

    PubMed  Google Scholar 

  45. Yi A, Klinman DM, Martin TL, Matson S, Krieg AM. Rapid immune activation by CpG motifs in bacterial DNA. J Immunol 157:5394–5402;1996.

    PubMed  Google Scholar 

  46. Yi A, Krieg AM. CpG DNA rescue from anti-IgM-induced WEHI-231 B lymphoma apoptosis via modulation of IκBα and IκBα and sustained activation of nuclear factor-κB/c-Rel. J Immunol 160:1240–1245;1998.

    PubMed  Google Scholar 

  47. Yi AK, Tuetken R, Redford T, Kirsch J, Krieg AM. CpG motifs in bacterial DNA activate leukocytes through the ph-dependent generation of reactive oxygen species. J Immunol 160:4755–4761;1998.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Veterans Affairs Medical Center, Department of Internal Medicine and Interdisciplinary Immunology Program, University of Iowa, College of Medicine, Iowa City, Iowa

    Mary Bradford, Arin J. Schroeder &  John S. Cowdery

  2. National Institute of Allergy and Infectious Diseases, National Institute of Health, USA

    Herbert C. Morse 3rd

  3. Department of Microbiology and Immunology, Uniformed Services University of Health Sciences, Bethesda, Md., USA

    Stefanie N. Vogel

Authors
  1. Mary Bradford
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Arin J. Schroeder
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Herbert C. Morse 3rd
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Stefanie N. Vogel
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. John S. Cowdery
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bradford, M., Schroeder, A.J., Morse, H.C. et al. CpG DNA induced IL-12 p40 gene activation is independent of STAT1 activation or production of interferon consensus sequence binding protein. J Biomed Sci 9, 688–696 (2002). https://doi.org/10.1007/BF02254997

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02254997

Key Words

Search

Navigation