Skip to main content

Advertisement

SpringerLink
Log in

Interleukin-32γ Enhances the Production of IL-6 and IL-8 in Fibroblast-Like Synoviocytes Via Erk1/2 Activation

  • Published:
Journal of Clinical Immunology Aims and scope Submit manuscript

Abstract

Introduction

In the present study, we examined the effect of the pro-inflammatory cytokine IL-32γ, the most biologically active isoform, and its related molecules in fibroblast-like synoviocytes (FLS).

Materials and Methods

FLS were isolated from synovial tissues of rheumatoid arthritis (RA) patients. The secretion and expression of IL-6 and IL-8 were examined by ELISA and real-time PCR, and the activation of signaling molecules was evaluated by Western blot, electrophoretic mobility shift assay (EMSA), real-time PCR, and siRNA transfection.

Results

By IL-32γ stimulation in RA FLS, the expressions of IL-6 and IL-8 were increased significantly, and the phosphorylated Erk1/2 and AP-1 were expressed prominently in Western blot and EMSA. In the Erk1/2 inhibited cells, IL-32γ stimulation did not increase the mRNA expression of IL-6 and IL-8.

Conclusion

Our results suggest that IL-32γ stimulation can induce the production of IL-6 and IL-8 from RA FLS via Erk1/2 activation.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Edwards JC. The nature and origins of synovium: experimental approaches to the study of synoviocyte differentiation. J Anat. 1994;184:493–501.

    PubMed  Google Scholar 

  2. Noss EH, Brenner MB. The role and therapeutic implications of fibroblast-like synoviocytes in inflammation and cartilage erosion in rheumatoid arthritis. Immunol Rev. 2008;223:252–70.

    Article  CAS  PubMed  Google Scholar 

  3. Kim KW, Cho ML, Kim HR, Ju JH, Park MK, Oh HJ, et al. Up-regulation of stromal cell-derived factor 1 (CXCL12) production in rheumatoid synovial fibroblasts through interactions with T lymphocytes: role of interleukin-17 and CD40L-CD40 interaction. Arthritis Rheum. 2007;56:1076–86.

    Article  CAS  PubMed  Google Scholar 

  4. Nanki T, Nagasaka K, Hayashida K, Saita Y, Miyasaka N. Chemokines regulate IL-6 and IL-8 production by fibroblast-like synoviocytes from patients with rheumatoid arthritis. J Immunol. 2001;167:5381–5.

    CAS  PubMed  Google Scholar 

  5. Kim SH, Han SY, Azam T, Yoon DY, Dinarello CA. Interleukin-32: a cytokine and inducer of TNFalpha. Immunity. 2005;22:131–42.

    CAS  PubMed  Google Scholar 

  6. Netea MG, Azam T, Ferwerda G, Girardin SE, Walsh M, Park JS, et al. IL-32 synergizes with nucleotide oligomerization domain (NOD) 1 and NOD2 ligands for IL-1beta and IL-6 production through a caspase 1-dependent mechanism. Proc Natl Acad Sci U S A. 2005;102:16309–14.

    Article  CAS  PubMed  Google Scholar 

  7. Goda C, Kanaji T, Kanaji S, Tanaka G, Arima K, Ohno S, et al. Involvement of IL-32 in activation-induced cell death in T cells. Int Immunol. 2006;18:233–40.

    Article  CAS  PubMed  Google Scholar 

  8. Choi JD, Bae SY, Hong JW, Azam T, Dinarello CA, Her E, et al. Identification of the most active interleukin-32 isoform. Immunology. 2009;126:535–42.

    Article  CAS  PubMed  Google Scholar 

  9. Joosten LA, Netea MG, Kim SH, Yoon DY, Oppers-Walgreen B, Radstake TR, et al. IL-32, a proinflammatory cytokine in rheumatoid arthritis. Proc Natl Acad Sci U S A. 2006;103:3298–303.

    Article  CAS  PubMed  Google Scholar 

  10. Kim YG, Lee CK, Oh JS, Kim SH, Kim KA, Yoo B. Effect of interleukin-32γ on differentiation of osteoclasts from CD14+ monocytes. Arthritis Rheum. 2010 (in press).

  11. Arnett FC, Edworthy SM, Bloch DA, McShane DJ, Fries JF, Cooper NS, et al. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum. 1988;31:315–24.

    Article  CAS  PubMed  Google Scholar 

  12. Takayanagi H, Oda H, Yamamoto S, Kawaguchi H, Tanaka S, Nishikawa T, et al. A new mechanism of bone destruction in rheumatoid arthritis: synovial fibroblasts induce osteoclastogenesis. Biochem Biophys Res Commun. 1997;240:279–86.

    Article  CAS  PubMed  Google Scholar 

  13. Johnson MR, Wang K, Smith JB, Heslin MJ, Diasio RB. Quantitation of dihydropyrimidine dehydrogenase expression by real-time reverse transcription polymerase chain reaction. Anal Biochem. 2000;278:175–84.

    Article  CAS  PubMed  Google Scholar 

  14. Schreiber E, Matthias P, Muller MM, Schaffner W. Rapid detection of octamer binding proteins with ‘mini-extracts’, prepared from a small number of cells. Nucleic Acids Res. 1989;17:6419.

    Article  CAS  PubMed  Google Scholar 

  15. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248–54.

    Article  CAS  PubMed  Google Scholar 

  16. Shoda H, Fujio K, Yamaguchi Y, Okamoto A, Sawada T, Kochi Y, et al. Interactions between IL-32 and tumor necrosis factor alpha contribute to the exacerbation of immune-inflammatory diseases. Arthritis Res Ther. 2006;8:R166.

    Article  PubMed  CAS  Google Scholar 

  17. Shioya M, Nishida A, Yagi Y, Ogawa A, Tsujikawa T, Kim-Mitsuyama S, et al. Epithelial overexpression of interleukin-32alpha in inflammatory bowel disease. Clin Exp Immunol. 2007;149:480–6.

    Article  CAS  PubMed  Google Scholar 

  18. Nishida A, Andoh A, Shioya M, Kim-Mitsuyama S, Takayanagi A, Fujiyama Y. Phosphatidylinositol 3-kinase/Akt signaling mediates interleukin-32alpha induction in human pancreatic periacinar myofibroblasts. Am J Physiol Gastrointest Liver Physiol. 2008;294:G831–8.

    Article  CAS  PubMed  Google Scholar 

  19. Mun SH, Kim JW, Nah SS, Ko NY, Lee JH, Kim JD, et al. Tumor necrosis factor alpha-induced interleukin-32 is positively regulated via the Syk/protein kinase Cdelta/JNK pathway in rheumatoid synovial fibroblasts. Arthritis Rheum. 2009;60:678–85.

    Article  CAS  PubMed  Google Scholar 

  20. Novick D, Rubinstein M, Azam T, Rabinkov A, Dinarello CA, Kim SH. Proteinase 3 is an IL-32 binding protein. Proc Natl Acad Sci U S A. 2006;103:3316–21.

    Article  CAS  PubMed  Google Scholar 

  21. Firestein GS, Manning AM. Signal transduction and transcription factors in rheumatic disease. Arthritis Rheum. 1999;42:609–21.

    Article  CAS  PubMed  Google Scholar 

  22. Neff L, Zeisel M, Sibilia J, Scholler-Guinard M, Klein JP, Wachsmann D. NF-kappaB and the MAP kinases/AP-1 pathways are both involved in interleukin-6 and interleukin-8 expression in fibroblast-like synoviocytes stimulated by protein I/II, a modulin from oral streptococci. Cell Microbiol. 2001;3:703–12.

    Article  CAS  PubMed  Google Scholar 

  23. Mabilleau G, Sabokbar A. Interleukin-32 promotes osteoclast differentiation but not osteoclast activation. PLoS ONE. 2009;4:e4173.

    Article  PubMed  CAS  Google Scholar 

  24. Garcia-Vicuna R, Gomez-Gaviro MV, Dominguez-Luis MJ, Pec MK, Gonzalez-Alvaro I, Alvaro-Gracia JM, et al. CC and CXC chemokine receptors mediate migration, proliferation, and matrix metalloproteinase production by fibroblast-like synoviocytes from rheumatoid arthritis patients. Arthritis Rheum. 2004;50:3866–77.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by grant from Asan Institute for Life Science (2008-330) and Basic Science Research Program through the National Research Foundation of Korea (NRF, 2009-0071413).

Author information

Authors and Affiliations

  1. Division of Rheumatology, Department of Internal Medicine, University of Ulsan College of Medicine, Asan Medical Center, 388-1 Pungnap-dong, Songpa-gu, 138-736, Seoul, South Korea

    Yong-Gil Kim, Chang-Keun Lee & Bin Yoo

  2. Department of Biomedical Science and Technology, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, 143-701, Seoul, South Korea

    Soo-Hyun Kim

  3. Department of Orthopaedic Surgery, University of Ulsan College of Medicine, Asan Medical Center, 388-1 Pungnap-dong, Songpa-gu, 138-736, Seoul, South Korea

    Woo-Shin Cho

  4. Department of Immunology and Physiology, College of Medicine, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, 143-701, Seoul, South Korea

    Se Hwan Mun

Authors
  1. Yong-Gil Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Chang-Keun Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Soo-Hyun Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Woo-Shin Cho
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Se Hwan Mun
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Bin Yoo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Chang-Keun Lee.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kim, YG., Lee, CK., Kim, SH. et al. Interleukin-32γ Enhances the Production of IL-6 and IL-8 in Fibroblast-Like Synoviocytes Via Erk1/2 Activation. J Clin Immunol 30, 260–267 (2010). https://doi.org/10.1007/s10875-009-9360-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10875-009-9360-2

Keywords

Search

Navigation