Skip to main content
Download PDF
  • Short Communication
  • Published:

The influence of JNK and P38 MAPK inhibition on IL-12P40 and IL-23 production depending on IL12B promoter polymorphism

Cellular & Molecular Biology Letters volume 14, pages 609–621 (2009)Cite this article

Abstract

The interleukin-12p40 gene (IL12B) encodes the p40 polypeptide chain, which, together with p19, composes IL-23. A bi-allelic promoter polymorphism (IL12Bpro) located at −2703 bp of the transcription initiation site has been reported to show associations with IL-12p40 production. To elucidate the dependence of IL-12p40 and IL-23 production on IL12Bpro polymorphism in relation to MAPK signal transduction pathways, we examined the effect of JNK and p38 inhibition on the secretion of these cytokines by stimulated peripheral blood mononuclear cells (PBMC) from healthy donors with 1.1 and 1.2/2.2 IL12Bpro genotypes. Stimulation with LPS and C3bgp resulted in approximately equal IL-12p40 production from PBMC with the 1.1 and 1.2/2.2 genotypes. The inhibition of JNK and p38 before stimulation significantly upregulated IL-12p40 production by PBMC with the 1.1 genotype, but did not influence IL-12p40 production from PBMC with the 1.2/2.2 genotype. Cultures of PBMC with the 1.1 genotype produced significantly more IL-12p40 than PBMC with the 1.2/2.2 genotype after stimulation with PHA. Inhibition of p38 kinase upregulated p40 production only in cultures with the 1.1 genotype. Decreased IL-23 production was observed in C3bgp-stimulated cultures after the inhibition of p38 regardless of the genotype of the tested cells. We concluded that IL-12p40 and IL-23 expression, which is mediated by the p38 and JNK intracellular signaling pathways, is influenced by IL12Bpro polymorphism.

Abbreviations

C3bgp:

C3 binding glycoprotein

IFN-γ:

interferon gamma

IL:

interleukin

IL12B :

the gene for p40 chain of IL-12

LPS:

lipopolysaccharide

MAPK:

mitogen-activated protein kinase

SAC:

Staphylococcus aureus cells

PBMC:

peripheral blood mononuclear cells

PHA:

phytohemagglutinin

References

  1. Hölscher, C. The power of combinatorial immunology: IL-12 and IL-12- related dimeric cytokines in infectious diseases. Med. Microbiol. Immunol. 193 (2004) 1–17.

    Article  PubMed  CAS  Google Scholar 

  2. Oppmann, B., Lesley, R., Blom, B., Timans, J.C., Xu, Y., Hunte, B., Vega, F., Yu, N., Wang, J., Singh, K., Zonin, F., Vaisberg, E., Churakova, T., Liu, M., Gorman, D., Wagner, J., Zurawski, S., Liu, Y., Abrams, J.S., Moore, K.W., Rennick, D., Waal-Malefyt, R., Hannum, C., Bazan, J.F., and Kastelein, R.A. Novel p19 protein engages IL-12p40 to form a cytokine, IL-23, with biological activities similar as well as distinct from IL-12. Immunity 13 (2000) 715–725.

    Article  PubMed  CAS  Google Scholar 

  3. Trinchieri, G. Interleukin-12: a proinflammatory cytokine with immunoregulatory functions that bridge innate resistance and antigen-specific adaptive immunity. Annu. Rev. Immunol. 13 (1995) 251–256.

    Article  PubMed  CAS  Google Scholar 

  4. Lankford, R.S.C. and Fricht, M.D. A unique role for IL-23 in promoting cellular immunity. J. Leukoc. Biol. 73 (2003) 49–56.

    Article  PubMed  CAS  Google Scholar 

  5. Gutcher, I. and Becher, B. APC-derived cytokines and T cell polarization in autoimmune inflammation. J. Clin. Invest. 117 (2007) 1119–1127.

    Article  PubMed  CAS  Google Scholar 

  6. Wysocka, M., Kubin, M., Vieira, L.Q., Ozmen, L., Garotta, G., Scott, P. and Trinchieri, G. Interleukin 12 is required for interferon-γ production and lethality in lipopolysaccharide-induced shock in mice. Eur. J. Immunol. 25 (1995) 672–676.

    Article  PubMed  CAS  Google Scholar 

  7. D’Andrea, A., Rengaraju, M., Valiante, N.M., Chehimi, J., Kubin, M., Aste, M., Chan, S.H., Kobayashi, M., Young, D., Nickbarg, E., Chizzonite, R., Wolf, S.F., and Trinchieri, G. Production of natural killer cell stimulatory factor (NKSF/IL-12) by peripheral blood mononuclear cells. J. Exp. Med. 176 (1992) 1387–1398.

    Article  PubMed  Google Scholar 

  8. Cooper, A.M. and Khader, S.A. IL-12p40: an inherently agonistic cytokine. Trends Immunol. 28 (2006) 33–38.

    Article  PubMed  CAS  Google Scholar 

  9. Morahan, G., Huang, D., Wu, M., Holt, B.J., White, G.P., Kendall, G.E., Sly, P.D., and Holt, P.G. Association of IL12B promoter polymorphism with severity of atopic and non-atopic asthma in children. Lancet 10 (2002) 455–459.

    Article  Google Scholar 

  10. Tatebayashi, K., Matsui, E., Kaneko, H., Fukao, T., Kasahara, K. and Kondo, N. IL-12B promoter polymorphism associated with asthma and IL-12B transcriptional activity. Allergol. Int. 54 (2005) 451–459.

    Article  CAS  Google Scholar 

  11. Marquet, S., Doumbo, O., Cabantous, S., Poudiougou, B., Argiro, L., Safeukui, I., Konate, S., Sissoko, S., Chevereau, E., Traore, A., Keita, M.M., Chevillard, C., Abel, L. and Dessein, A.J. A functional promoter variant in IL12B predisposes to cerebral malaria. Hum. Mol. Genet. 17 (2008) 2190–2195.

    Article  PubMed  CAS  Google Scholar 

  12. Morahan, G., Huang, D., Ymer, S.I., Cancilla, M.R., Stephen, K., Dabadghao, P., Werther, G., Tait, B.D., Harrison, L.C. and Colman, P.G. Linkage disequilibrium of type 1 diabetes susceptibility locus with a regulatory IL12B allele. Nat. Genet. 27 (2001) 218–221.

    Article  PubMed  CAS  Google Scholar 

  13. Utsugi, M., Dobashi, K., Ishizuka, T., Endou, K., Hamuro, J., Murata, Y., Nakazava, T., and Mori, M., c-Jun N-terminal kinase negatively regulates lipopolysaccharide-induced IL-12 production in human macrophages: role of mitogen-activated protein kinase in glutathione redox regulation of IL-12 production. J. Immunol. 171 (2003) 628–635.

    PubMed  CAS  Google Scholar 

  14. Ma, W., Gee, K., Lim, W., Chambers, K., Angel, J.B., Kozlowski, M., and Kumar, A., Dexamethasone inhibits IL-12p40 production in lipopolysaccharide-stimulated human monocytic cells by down-regulating the activity of c-Jun N-terminal kinase, the Activation protein-1 and NF-κB transcription factors. J. Immunol. 172 (2004) 318–330.

    PubMed  CAS  Google Scholar 

  15. Petro, T. M. ERK-MAP-kinases differentially regulate expression of IL-23p19 compared with p40 and IFN-beta in Theiler’s virus-infected RAW264.7 cells. Immunol. Lett. 97 (2005) 47–53.

    Article  PubMed  CAS  Google Scholar 

  16. Zhelev, Z., Stanilova, S. and Carpenter, B. Isolation, partial characterization and complement inhibiting activity of a new glycoprotein from Cuscuta europea. Biochem. Biophys. Res. Commun. 202 (1994) 186–194.

    Article  PubMed  CAS  Google Scholar 

  17. Stanilova, S., Zhelev, Zh. and Dobreva, Z. Preliminary studies on the immunomodulatory effect of the C3 binding glycoprotein isolated from Cuscuta europea. Int. J. Immunopharmacol. 22 (2000) 15–24.

    Article  PubMed  CAS  Google Scholar 

  18. Stanilova, S.A., Dobreva, Z.G., Slavov, E.S. and Miteva, L.D. C3 binding glycoprotein from Cuscuta europea induced different cytokine profiles from human PBMC compared to other plant and bacterial immunomodulators. Int. Immunopharmacol. 5 (2005) 723–734.

    Article  PubMed  CAS  Google Scholar 

  19. Dobreva, Z.G. and Stanilova, S.A. The immunomodulatory activity of C3 binding glycoprotein (C3bgp) is mediated by the complement receptor type III and mitogen-activated protein kinase signal transduction pathways. Immunotoxicol. 29 (2007) 1–14.

    Google Scholar 

  20. Dobreva, Z.G., Stanilova, S.A. and Miteva, L.D. Differences in the inducible gene expression and protein production of IL-12p40, IL-12p70 and IL-23: Involvement of p38 and JNK kinase pathways. Cytokine 43 (2008) 76–82.

    Article  PubMed  CAS  Google Scholar 

  21. Rao, K.M.K. MAP kinase activation in macrophages. J. Leukoc. Biol. 69 (2001) 3–10.

    PubMed  CAS  Google Scholar 

  22. Bennet, B.L., Sasaki, D.T., Murray, B.W., O’Leary, E.C., Sakata, S.T., Xu, W., Leisten, J.C., Motiwata, A., Pierce, S., Satoh, Y., Bhagwat, S.S., Manning, A.M., and Anderson, D.W. SP600125, an anthrapyrazolone inhibitor of Jun N-terminal kinase. Proc. Natl. Acad. Sci. USA 98 (2001) 13681–13686.

    Article  Google Scholar 

  23. Manthey, C.L., Wang, S.W., Kinney, S.D., and Yao, Z. SB202190, a selective inhibitor of p38 mitogen-activated protein kinase, is a powerful regulator of LPS-induced mRNAs in monocytes. J. Leukoc. Biol. l64 (1998) 409–417.

    Google Scholar 

  24. Müller-Berghaus, J., Kern, K., Pshen, A., Nguyen, X.D., Kluter, H., Morahan, G., and Schadendorf, D. Deficient IL-12p70 secretion by dendritic cells based on IL12B promoter genotype. Genes Immun. 5 (2004) 431–434.

    Article  PubMed  CAS  Google Scholar 

  25. Stanilova, S., Miteva, L. and Prakova, G. IL-12Bpro and GSTP1 polymorphisms in association with silicosis. Tissue Antigens 71 (2007) 169–174.

    Article  PubMed  Google Scholar 

  26. Marriott, J.B., Clarke, I.A. and Dalgleish, A.G. Inhibition of p38 MAP kinase during cellular activation results in IFN-c-dependent augmentation of IL-12 production by human monocytes/macrophages. Clin. Exp. Immunol. 125 (2001) 64–70.

    Article  PubMed  CAS  Google Scholar 

  27. Yoshimoto, T., Wang, C.R., Yoneto, T., Waki, S., Sunaga, S., Komagata, Y., Mitsuyama, M., Miyazaki, J., and Nariuchi, H. Reduced T helper 1 responses in IL-12p40 transgenic mice. J. Immunol. 160 (1998) 588–594.

    PubMed  CAS  Google Scholar 

  28. Sommer, S., Pudrith, C.B., Colvin, C.J., Coussens, C.P. Mycobacterium avium subspecies paratuberculosis suppresses expression of IL-12p40 and iNOS genes induced by signaling through CD40 in bovine monocyte-derived macrophages. Vet. Immunol. Immunopthol. 128 (2009) 44–52.

    Article  CAS  Google Scholar 

  29. Ma, X., Chow, J.M., Gri, G., Carra, G., Gerosa, F., Wolf, S.F., Dzialo, R. and Trinchieri, G. The Interleukin 12 p40 gene promoter is primed by Interferon γ in monocytic cells. J. Exp. Med. 183 (1996) 147–157.

    Article  PubMed  CAS  Google Scholar 

  30. Hino, A., Kweon, M.N., Fujihashi, K., McGhee, J.R., and Kiyono, H. Pathological role of large intestinal IL-12p40 for the induction of Th2-type allergic diarrhea. Am. J. Pathol. 164 (2004) 1327–1335.

    PubMed  CAS  Google Scholar 

  31. Walter, M.J., Kajivara, N., Karanja, P., Castro, M., and Holtzman, M.J. Interlekin 12p40 production by barrier epithelial cells during airway inflammation. J. Exp. Med. 193 (2001) 339–351.

    Article  PubMed  CAS  Google Scholar 

  32. Khader, S.A., Partida-Sanchez, S., Bell, G., Jelley-Gibbs, D.M., Swain, S., Pearl, J.E., Ghilardi, N., deSauvage, F.J., Lund, F.E., and Cooper, A.M. Interleukin 12p40 is required for dendritic cell migration and T cell priming after Mycobacterium tuberculosis infection. J. Exp. Med. 203 (2006) 1805–1815.

    Article  PubMed  CAS  Google Scholar 

  33. Kim, D.J., Youn, J.I., Seo, S.H., Jin, H.T. and Sung, Y.S. Differential regulation of antigen-specific CD8+ T cell responses by IL-12p40 in a dose dependent manner. J. Immunol. 180 (2008) 7167–7174.

    PubMed  CAS  Google Scholar 

  34. Ma, X., Neurath, M., Gri, G. and 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 (1997) 10389–10395.

    Article  PubMed  CAS  Google Scholar 

  35. Murphy, T.L., Cleveland, M.G., Kulesza, P., Magram, J. and Murphy, K.M. Regulation of interleukin 12 p40 expression through an NF-kappa B half-site. Mol. Cell. Biol. 15 (1995) 5258–5267.

    PubMed  CAS  Google Scholar 

  36. Plevy, S.E., Gemberling, J.H., Hsu, S., Dorner, A.J. and Smale, S.T. 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 (1997) 4572–4588.

    PubMed  CAS  Google Scholar 

  37. Zhu, C., Gagnidze, K., Gemberling, J.H. and Plevy, S.E. Characterization of an activation protein-1-binding site in the murine interleukin-12 p40 promoter. J. Biol. Chem. 276 (2001) 18519–18528.

    Article  PubMed  CAS  Google Scholar 

  38. Mitsuhashi, M., Liu, J., Cao, S., Shi, X. and Ma, X. Regulation of interleukin-12 gene expression and its anti-tumor activities by prostaglandin E2 derived from mammary carcinomas. J. Leukoc. Biol. 76 (2004) 322–332.

    Article  PubMed  CAS  Google Scholar 

  39. Karin, M. the regulation of AP-1 activity by mitogen-activated protein kinases. J. Biol. Chem. 270 (1995) 16483–16486.

    PubMed  CAS  Google Scholar 

  40. Tanos, T., Marinissen, M.J., Leskow, F.C. Hochbaum, D., Martinetto, H., Gutkind, S. and Coso, O.A. Phosphorylation of c-Fos by members of the p38 MAPK family. J. Biol. Chem. 280 (2005) 18842–18852.

    Article  PubMed  CAS  Google Scholar 

  41. Bhat, N.R., Feinstein, D.L., Shen, A.N., Bhat, A.N. p38-mediated transcriptional activation of inducible nitric-oxide synthase in glial cells. Roles of nuclear factors, nuclear factor kappa B, cAMP response element-binding protein, CCAAT/enhancer-binding protein-beta, and activating transcription factor-2. J. Biol. Chem. 277 (2002) 29584–29592.

    Article  PubMed  CAS  Google Scholar 

  42. Shimokawa, N., Nishiyama, C., Hirota, T., Hara, M., Ikeda, S., Okumura, K. and Ogawa, H. Functional analysis of a polymorphism in the promoter region of the IL-12/23p40 gene. Clin. Exp. Allergy 39 (2009) 228–235.

    Article  PubMed  CAS  Google Scholar 

  43. D’Addario, M., Arora, P.D., McCulltoch, C.A. role of p38 in stress activation of Sp1. Gene 379 (2006) 51–61.

    Article  PubMed  CAS  Google Scholar 

  44. Sheikh, S., Matsuoka, K., Li, F. and Plevy, S. Molecular regulation of IL-23 p19 gene expression in macrophages and murine inflammatory bowel disease. Clin. Immunol. 127Suppl.1 (2008) S119.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Molecular Biology, Immunology & Genetics, Faculty of Medicine, Trakia University, Armeiska 11 St., 6000, Stara Zagora, Bulgaria

    Zlatka Georgieva Dobreva, Spaska Angelova Stanilova & Lyuba Dineva Miteva

Authors
  1. Zlatka Georgieva Dobreva
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Spaska Angelova Stanilova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lyuba Dineva Miteva
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zlatka Georgieva Dobreva.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dobreva, Z.G., Stanilova, S.A. & Miteva, L.D. The influence of JNK and P38 MAPK inhibition on IL-12P40 and IL-23 production depending on IL12B promoter polymorphism. Cell Mol Biol Lett 14, 609–621 (2009). https://doi.org/10.2478/s11658-009-0022-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2478/s11658-009-0022-4

Key words

Cellular & Molecular Biology Letters

ISSN: 1689-1392

Contact us