Skip to main content
SpringerLink
Log in

Production of recombinant human interleukin-38 and its inhibitory effect on the expression of proinflammatory cytokines in THP-1 cells

  • Molecular Cell Biology
  • Published:
Molecular Biology Aims and scope Submit manuscript

Abstract

Interleukin (IL)-38 is the latest member of the IL-1 cytokine family. However, as a result of lacking efficient method to generate relatively large quantity of IL-38, its precise functions are poorly understood. In the present study, the cloning, expression, purification, and activity analysis of recombinant human IL-38 was described. Human IL-38 cDNA was cloned into the prokaryotic expression vector pET-44. The recombinant IL-38 containing a C-hexahistidine tag was expressed in Escherichia coli BL21 (DE3) which induced by isopropyl-β-D-thiogalactoside. The expressed fusion protein was purified by Ni-NTA affinity chromatography. IL-38 protein was largely found in the soluble fraction. The purified IL-38 appeared a single band on SDS-PAGE, the yield of IL-38 was 4 mg from 1 L of bacterial culture, and the purity was more than 98% with low endotoxin level (<0.1 EU/μg). Western blotting confirmed the identity of the purified protein. Activity analysis showed that IL-38 can inhibit effectively the expression of proinflammatory cytokines, such as tumor necrosis factor-α, IL-1β, IL-17, and monocyte chemoattractant protein-1 in lipopolysaccharide-activated THP-1 cells. The production and characterization of biologically active IL-38 will be beneficial for its potential role in clinical applications.

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. van de Veerdonk F.L., Stoeckman A.K., Wu G., Boeckermann A.N., Azam T., Netea M.G., Joosten L.A., van der Meer J.W., Hao R., Kalabokis V., Dinarello C.A. 2012. IL-38 binds to the IL-36 receptor and has biological effects on immune cells similar to IL-36 receptor antagonist. Proc. Natl. Acad. Sci. U. S. A. 109, 3001–3005.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Dinarello C.A. 2011. Interleukin-1 in the pathogenesis and treatment of inflammatory diseases. Blood. 117, 3720–3732.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Dinarello C., Arend W., Sims J., Smith D., Blumberg H., O’ Neill L., Goldbach-Mansky R., Pizarro T., Hoffman H., Bufler P., Nold M., Ghezzi P., Mantovani A., Garlanda C., Boraschi D., et al. 2010. IL-1 family nomenclature. Nat. Immunol. 11, 973.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Nicklin M.J., Barton J.L., Nguyen M., FitzGerald M.G., Duff G.W., Kornman K. 2002. A sequence-based map of the nine genes of the human interleukin-1 cluster. Genomics. 79, 718–725.

    Article  CAS  PubMed  Google Scholar 

  5. Bensen J.T., Dawson P.A., Mychaleckyj J.C., Bowden D.W. 2001. Identification of a novel human cytokine gene in the interleukin gene cluster on chromosome 2q12-14. J. Interferon Cytokine Res. 21, 899–904.

    Article  CAS  PubMed  Google Scholar 

  6. Kumar S., McDonnell P.C., Lehr R., Tierney L., Tzimas M.N., Griswold D.E., Capper E.A., Tal-Singer R., Wells G.I., Doyle M.L., Young P.R. 2000. Identification and initial characterization of four novel members of the interleukin-1 family. J. Biol. Chem. 275, 10308–10314.

    Article  CAS  PubMed  Google Scholar 

  7. Lin H., Ho A.S., Haley-Vicente D., Zhang J., Bernal- Fussell J., Pace A.M., Hansen D., Schweighofer K., Mize N.K., Ford J.E. 2001. Cloning and characterization of IL-1HY2, a novel interleukin-1 family member. J. Biol. Chem. 276, 20597–20602.

    Article  CAS  PubMed  Google Scholar 

  8. Shen H., Goodall J.C., Hill Gaston J.S. 2009. Frequency and phenotype of peripheral blood Th17 cells in ankylosing spondylitis and rheumatoid arthritis. Arthritis Rheum. 60, 1647–1656.

    Article  CAS  PubMed  Google Scholar 

  9. Hur W., Cho M.L., Yoon S.K., Kim S.Y., Ju J.H., Jhun J.Y., Heo S.B., Moon Y.M., Min S.Y., Park S.H., Kim H.Y. 2006. Adenoviral delivery of IL-1 receptor antagonist abrogates disease activity during the development of autoimmune arthritis in IL-1 receptor antagonist-deficient mice. Immunol. Lett. 106, 154–162.

    Article  CAS  PubMed  Google Scholar 

  10. Farooq M., Nakai H., Fujimoto A., Fujikawa H., Matsuyama A., Kariya N., Aizawa A., Fujiwara H., Ito M., Shimomura Y. 2013. Mutation analysis of the IL36RN gene in 14 Japanese patients with generalized pustular psoriasis. Hum. Mutat. 34, 176–183.

    Article  CAS  PubMed  Google Scholar 

  11. Hu Z., Chen Z., Huang N., Teng X., Zhang J., Wang Z., Wei X., Qin K., Liu X., Wu X., Tang H., Zhu X., Cui K., Li J. 2015. Expression, purification of IL-38 in Escherichia coli and production of polyclonal antibodies. Protein Expr. Purif. 107, 76–82.

    Article  CAS  PubMed  Google Scholar 

  12. Cheng X., Shen Y., Li R. 2014. Targeting TNF: A therapeutic strategy for Alzheimer’s disease. Drug Discov. Today. 19, 1822–1827.

    Article  CAS  PubMed  Google Scholar 

  13. Singhal G., Jaehne E.J., Corrigan F., Toben C., Baune B.T. 2014. Inflammasomes in neuroinflammation and changes in brain function: A focused review. Front. Neurosci. 8, 315.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Tam L.S., Kitas G.D., Gonzalez-Gay M.A. 2014). Can suppression of inflammation by anti-TNF prevent progression of subclinical atherosclerosis in inflammatory arthritis? Rheumatology (Oxford). 53, 1108–1119.

  15. Qamar A., Rader D.J. 2012. Effect of interleukin 1beta inhibition in cardiovascular disease. Curr. Opin. Lipidol. 23, 548–553.

    Article  CAS  PubMed  Google Scholar 

  16. Kim K.O., Jang B.I. 2011. Emerging drugs in the treatment of inflammatory bowel disease: Beyond anti- TNF-alpha. Korean J. Gastroenterol. 58, 235–244.

    Article  PubMed  Google Scholar 

  17. Peyrin-Biroulet L. 2010. Anti-TNF therapy in inflammatory bowel diseases: A huge review. Minerva Gastroenterol. Dietol. 56, 233–243.

    CAS  PubMed  Google Scholar 

  18. Aguilera M., Darby T., Melgar S. 2014. The complex role of inflammasomes in the pathogenesis of Inflammatory bowel diseases: Lessons learned from experimental models. Cytokine Growth Factor Rev. 25, 715–730.

    Article  CAS  PubMed  Google Scholar 

  19. Simsek I. 2011. TNF inhibitors for rheumatoid arthritis: A year in review. Bull. N.Y.U. Hosp. Jt. Dis. 69, 220–224.

    Google Scholar 

  20. Sethi G., Sung B., Aggarwal B.B. 2008. TNF: A master switch for inflammation to cancer. Front. Biosci. 13, 5094–5107.

    Article  CAS  PubMed  Google Scholar 

  21. Hundsberger H., Verin A., Wiesner C., Pfluger M., Dulebo A., Schutt W., Lasters I., Mannel D.N., Wendel A., Lucas R. 2008. TNF: A moonlighting protein at the interface between cancer and infection. Front. Biosci. 13, 5374–5386.

    Article  CAS  PubMed  Google Scholar 

  22. Kono H., Kimura Y., Latz E. 2014. Inflammasome activation in response to dead cells and their metabolites. Curr. Opin. Immunol. 30C, 91–98.

    Article  Google Scholar 

  23. Mikos H., Mikos M., Rabska-Pietrzak B., Niedziela M. 2014. The clinical role of serum concentrations of selected cytokines: IL-1beta, TNF-alpha and IL-6 in diagnosis of autoimmune thyroid disease (AITD) in children. Autoimmunity. 47, 466–472.

    Article  CAS  PubMed  Google Scholar 

  24. Gottlieb P.A., Alkanani A.K., Michels A.W., Lewis E.C., Shapiro L., Dinarello C.A., Zipris D. 2014. alpha1- Antitrypsin therapy downregulates toll-like receptorinduced IL-1beta responses in monocytes and myeloid dendritic cells and may improve islet function in recently diagnosed patients with type 1 diabetes. J. Clin. Endocrinol. Metab. 99, e1418–E1426.

    Article  Google Scholar 

  25. Mandolesi G., Gentile A., Musella A., Centonze D. 2015. IL-1beta dependent cerebellar synaptopathy in a mouse mode of multiple sclerosis. Cerebellum. 14, 19–22.

    Article  CAS  PubMed  Google Scholar 

  26. Wang Y., Gao B., Xiong S. 2014. Involvement of NLRP3 inflammasome in CVB3-induced viral myocarditis. Am. J. Physiol. Heart Circ. Physiol. 307, H1438–H1447.

    Article  CAS  PubMed  Google Scholar 

  27. Saghazadeh A., Mastrangelo M., Rezaei N. 2014. Genetic background of febrile seizures. Rev. Neurosci. 25, 129–161.

    CAS  PubMed  Google Scholar 

  28. Lorigados Pedre L., Morales Chacon L. M., Orozco Suarez S., Pavon Fuentes N., Estupinan Diaz B., Serrano Sanchez T., Garcia Maeso I., Rocha Arrieta L. 2013. Inflammatory mediators in epilepsy. Curr. Pharm. Des. 19, 6766–6772.

    Article  PubMed  Google Scholar 

  29. Harrold L. 2013. New developments in gout. Curr. Opin. Rheumatol. 25, 304–309.

    Article  CAS  PubMed  Google Scholar 

  30. Zambrano-Zaragoza J.F., Romo-Martinez E.J., Duran-Avelar Mde J., Garcia-Magallanes N., Vibanco-Perez N. 2014. Th17 cells in autoimmune and infectious diseases. Int. J. Inflam. 2014, 651503.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Yang J., Sundrud M.S., Skepner J., Yamagata T. 2014. Targeting Th17 cells in autoimmune diseases. Trends Pharmacol. Sci. 35, 493–500.

    Article  CAS  PubMed  Google Scholar 

  32. Kawano S., Nakamachi Y. 2011. miR-124a as a key regulator of proliferation and MCP-1 secretion in synoviocytes from patients with rheumatoid arthritis. Ann. Rheum. Dis. 70 (Suppl. 1), 88–91.

    Article  Google Scholar 

  33. Evans A.C., Papachristou G.I., Whitcomb D.C. 2010). Obesity and the risk of severe acute pancreatitis. Minerva Gastroenterol. Dietol. 56, 169–179.

    CAS  Google Scholar 

  34. Panee J. 2012. Monocyte chemoattractant protein 1 (MCP-1) in obesity and diabetes. Cytokine. 60, 1–12.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Rose C.E., Sung S.S., Fu S.M. 2003. Significant involvement of CCL2 (MCP-1) in inflammatory disorders of the lung. Microcirculation. 10, 273–288.

    Article  CAS  PubMed  Google Scholar 

  36. Matoba T., Egashira K. 2011. Anti-inflammatory gene therapy for cardiovascular disease. Curr. Gene Ther. 11, 442–446.

    Article  CAS  PubMed  Google Scholar 

  37. Tete S., Nicoletti M., Saggini A., Maccauro G., Rosati M., Conti F., Cianchetti E., Tripodi D., Toniato E., Fulcheri M., Salini V., Caraffa A., Antinolfi P., Frydas S., Pandolfi F., et al. 2012. Nutrition and cancer prevention. Int. J. Immunopathol. Pharmacol. 25, 573–581.

    CAS  PubMed  Google Scholar 

  38. Czaja A.J. 2014. Review article: Chemokines as orchestrators of autoimmune hepatitis and potential therapeutic targets. Aliment. Pharmacol. Ther. 40, 261–279.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Zhejiang Provincial Key Laboratory of Pathophysiology, Department of Immunology, Ningbo University School of Medicine, Ningbo, 315211, China

    X. L. Yuan, Y. Li, X. H. Pan, M. Zhou, Q. Y. Gao & M. C. Li

Authors
  1. X. L. Yuan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Y. Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. X. H. Pan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Q. Y. Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M. C. Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Y. Li or M. C. Li.

Additional information

Published in Russian in Molekulyarnaya Biologiya, 2016, Vol. 50, No. 3, pp. 466–473.

The article is published in the original.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yuan, X.L., Li, Y., Pan, X.H. et al. Production of recombinant human interleukin-38 and its inhibitory effect on the expression of proinflammatory cytokines in THP-1 cells. Mol Biol 50, 405–411 (2016). https://doi.org/10.1134/S0026893316030134

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S0026893316030134

Keywords

Search

Navigation