Skip to main content
NCBI home page
Annals of the Rheumatic Diseases logoLink to Annals of the Rheumatic Diseases
. 2004 Jan;63(1):43–49. doi: 10.1136/ard.2002.005546

Interferon ß stimulates interleukin 1 receptor antagonist production in human articular chondrocytes and synovial fibroblasts

G Palmer 1, F Mezin 1, C Juge-Aubry 1, C Plater-Zyberk 1, C Gabay 1, P Guerne 1
PMCID: PMC1754734  PMID: 14672890

Abstract

Background: Interferon (IFN) ß displays anti-inflammatory and immunosuppressive activity and has been considered for the treatment of rheumatoid arthritis (RA). Information about the effects of this molecule on joint cells is scarce, however.

Objective: To investigate the effects of IFNß on the production of interleukin-1 receptor antagonist (IL1Ra) in human articular chondrocytes and synovial fibroblasts.

Methods: Chondrocytes and synovial fibroblasts were stimulated with IFNß alone or in combination with interleukin (IL) 1ß. IL1Ra concentrations in culture supernatants and cell lysates were determined by ELISA. Expression of mRNA encoding the secreted sIL1Ra or the intracellular icIL1Ra1 isoforms was quantified by real time reverse transcriptase-polymerase chain reaction.

Results: In chondrocytes, IFNß alone had no effect, but dose dependently enhanced the secretion of IL1Ra induced by IL1ß. Chondrocyte cell lysates contained undetectable or low levels of IL1Ra, even after stimulation with IL1ß and IFNß. Consistently, IL1ß and IFNß induced sIL1Ra mRNA expression in chondrocytes, while expression of icIL1Ra1 was not detectable. Human articular chondrocytes thus mainly produce secreted IL1Ra. In synovial fibroblasts, IFNß alone dose dependently increased IL1Ra secretion. In addition, IFNß enhanced the stimulatory effect of IL1ß on IL1Ra production. In synovial cell lysates, IFNß and IL1ß also increased IL1Ra levels. Consistently, IFNß and IL1ß induced the expression of both sIL1Ra and icIL1Ra1 mRNA in synovial fibroblasts.

Conclusion: IFNß increases IL1Ra production in joint cells, which may be beneficial in cartilage damaging diseases such as RA or osteoarthritis.

Full Text

The Full Text of this article is available as a PDF (466.6 KB).

Figure 1 .

Figure 1

Open in a new tab

Expression of IFNAR1 and IFNAR2 mRNA in human articular chondrocytes and synovial fibroblasts. Expression of mRNA encoding (A) IFNAR1 and (B) IFNAR2 was assessed by RT-PCR using total RNA from freshly isolated (lanes 1, 6, 7) or dedifferentiated (passage 1; lanes 4 and 5) human articular chondrocytes or from human synovial fibroblasts (passage 3; lanes 2 and 3). Cells were left unstimulated (lanes 1, 2, 4, 6) or stimulated for 24 hours with 1 ng/ml IL1ß (lanes 3, 5, 7). M: DNA molecular weight marker. Lane 8: distilled water was used as a negative control for PCR amplification.

Figure 2 .

Figure 2

Open in a new tab

Dose dependent effects of IFNß and IL1ß on IL1Ra secretion by human articular chondrocytes. (A) Human articular chondrocytes freshly isolated from OA knee joint cartilage or (B) dedifferentiated human articular chondrocytes (passage 4, OA femoral head) were left unstimulated (control; white bars) or stimulated with indicated doses of IFNß (grey bars; 250 U/ml corresponds to 1 ng/ml), 1 ng/ml IL1ß (dark grey bars), or the combination of 1 ng/ml IL1ß and indicated doses of IFNß (black bars) for 72 hours. (C) Freshly isolated human articular chondrocytes (OA knee) were left unstimulated (control; white bars) or stimulated with indicated doses of IL1ß (dark grey bars), 2500 U/ml (10 ng/ml) IFNß (grey bar) or the combination of 2500 U/ml (10 ng/ml) IFNß and indicated doses of IL1ß (black bars) for 72 hours. IL1Ra concentrations in culture supernatants were measured by ELISA. Each bar represents the mean (SEM) of three determinations in a representative experiment. * p<0.05 as compared with control; & p<0.01 as compared with cells treated with the same dose of IL1ß alone.

Figure 3 .

Figure 3

Open in a new tab

Effects of IFNß and IL1ß on the production of secreted and intracellular IL1Ra by in human articular chondrocytes. (A) Human articular chondrocytes freshly isolated from normal adult femoral head cartilage or (B) dedifferentiated human articular chondrocytes (passage 5, OA femoral head) were left unstimulated (control; white bars) or stimulated with 2500 U/ml (10 ng/ml) IFNß (grey bars), 1 ng/ml IL1ß (dark grey bars), or the combination of 2500 U/ml (10 ng/ml) IFNß and1 ng/ml IL1ß (black bars) for 72 hours. IL1Ra concentrations in culture supernatants and cell lysates were measured by ELISA. Each bar represents the mean (SEM) of three determinations in a representative experiment. * p<0.05 as compared with control; & p<0.001 as compared with cells treated with IL1ß alone.

Figure 4 .

Figure 4

Open in a new tab

Effects of IFNß and IL1ß on the expression of sIL1Ra mRNA in human articular chondrocytes. Expression of mRNA encoding sIL1Ra was quantified by real time RT-PCR using total RNA from dedifferentiated human articular chondrocytes (passages 1–3). Cells were left unstimulated (control) or stimulated for 18 hours with 2500 U/ml (10 ng/ml) IFNß (IFN), 1 ng/ml IL1ß (IL1) or the combination of 2500 U/ml (10 ng/ml) IFNß and 1 ng/ml IL1ß (IL1 + IFN). The amount of 28S rRNA was monitored as an internal control. The expression of sIL1Ra was corrected for 28S rRNA levels and the IL1Ra/28S ratios were normalised to the maximal value observed in each experiment, which was set to 100%. The results shown represent the mean (SEM) of data obtained with three samples from different donors (one normal adult femoral head, one normal paediatric vertebra, and one OA femoral head).

Figure 5 .

Figure 5

Open in a new tab

Dose dependent effects of IFNß and IL1ß on IL1Ra production in human synovial fibroblasts. (A) Human OA synovial fibroblasts (passage 4) were left unstimulated (control; white bars) or stimulated with indicated doses of IFNß (grey bars; 250 U/ml corresponds to 1 ng/ml), 1 ng/ml IL1ß (dark grey bar), or the combination of 1 ng/ml IL1ß and indicated doses of IFNß (black bars) for 72 hours. (B) Human OA synovial fibroblasts (passage 5) were left unstimulated (control; white bars) or stimulated with indicated doses of IL1ß (dark grey bars), 2500 U/ml (10 ng/ml) IFNß (grey bar), or the combination of 2500 U/ml (10 ng/ml) IFNß and indicated doses of IL1ß (black bars) for 72 hours. IL1Ra concentrations in culture supernatants were measured by ELISA. Each bar represents the mean (SEM) of three determinations in a representative experiment. * p<0.05 as compared with control; & p<0.05 as compared with cells treated with the same dose of IL1ß alone.

Figure 6 .

Figure 6

Open in a new tab

Effects of IFNß and IL1ß on the production of secreted and intracellular IL1Ra in human synovial fibroblasts. Human RA synovial fibroblasts (passage 5) were left unstimulated (control; white bars) or stimulated with 2500 U/ml (10 ng/ml) IFNß (grey bars), 1 ng/ml IL1ß (dark grey bars), or the combination of 2500 U/ml (10 ng/ml) IFNß and 1 ng/ml IL1ß (black bars) for 72 hours. IL1Ra concentrations in culture supernatants and cell lysates were measured by ELISA. Each bar represents the mean (SEM) of three determinations in a representative experiment. * p<0.05 as compared with control; & p<0.05 as compared with cells treated with IL1ß alone.

Figure 7 .

Figure 7

Open in a new tab

Effects of IFNß and IL1ß on the expression of sIL1Ra and icIL1Ra1 mRNA in human synovial fibroblasts. Expression levels of mRNA encoding (A) sIL1Ra and (B) icIL1Ra1 were quantified by real time RT-PCR using total RNA from human synovial fibroblasts (passages 2–14). Cells were left unstimulated (control) or stimulated for 18 hours with 2500 U/ml (10 ng/ml) IFNß (IFN), 1 ng/ml IL1ß (IL1) or the combination of 2500 U/ml (10 ng/ml) IFNß and 1 ng/ml IL1ß (IL1 + IFN). The amount of 28S rRNA was monitored as an internal control. The expression of sIL1Ra or icIL1Ra1 was corrected for 28S rRNA levels and the IL1Ra/28S ratios were normalised to the maximal value observed in each experiment, which was set to 100%. The results shown represent the mean (SEM) of data obtained with three samples from different donors (one OA and two RA samples).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arend W. P., Malyak M., Guthridge C. J., Gabay C. Interleukin-1 receptor antagonist: role in biology. Annu Rev Immunol. 1998;16:27–55. doi: 10.1146/annurev.immunol.16.1.27. [DOI] [PubMed] [Google Scholar]
  2. Chabaud M., Page G., Miossec P. Enhancing effect of IL-1, IL-17, and TNF-alpha on macrophage inflammatory protein-3alpha production in rheumatoid arthritis: regulation by soluble receptors and Th2 cytokines. J Immunol. 2001 Nov 15;167(10):6015–6020. doi: 10.4049/jimmunol.167.10.6015. [DOI] [PubMed] [Google Scholar]
  3. Coclet-Ninin J., Dayer J. M., Burger D. Interferon-beta not only inhibits interleukin-1beta and tumor necrosis factor-alpha but stimulates interleukin-1 receptor antagonist production in human peripheral blood mononuclear cells. Eur Cytokine Netw. 1997 Dec;8(4):345–349. [PubMed] [Google Scholar]
  4. Dripps D. J., Brandhuber B. J., Thompson R. C., Eisenberg S. P. Interleukin-1 (IL-1) receptor antagonist binds to the 80-kDa IL-1 receptor but does not initiate IL-1 signal transduction. J Biol Chem. 1991 Jun 5;266(16):10331–10336. [PubMed] [Google Scholar]
  5. Fernandes J., Tardif G., Martel-Pelletier J., Lascau-Coman V., Dupuis M., Moldovan F., Sheppard M., Krishnan B. R., Pelletier J. P. In vivo transfer of interleukin-1 receptor antagonist gene in osteoarthritic rabbit knee joints: prevention of osteoarthritis progression. Am J Pathol. 1999 Apr;154(4):1159–1169. doi: 10.1016/S0002-9440(10)65368-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gabay C., Smith M. F., Eidlen D., Arend W. P. Interleukin 1 receptor antagonist (IL-1Ra) is an acute-phase protein. J Clin Invest. 1997 Jun 15;99(12):2930–2940. doi: 10.1172/JCI119488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Granowitz E. V., Clark B. D., Mancilla J., Dinarello C. A. Interleukin-1 receptor antagonist competitively inhibits the binding of interleukin-1 to the type II interleukin-1 receptor. J Biol Chem. 1991 Aug 5;266(22):14147–14150. [PubMed] [Google Scholar]
  8. Hannum C. H., Wilcox C. J., Arend W. P., Joslin F. G., Dripps D. J., Heimdal P. L., Armes L. G., Sommer A., Eisenberg S. P., Thompson R. C. Interleukin-1 receptor antagonist activity of a human interleukin-1 inhibitor. Nature. 1990 Jan 25;343(6256):336–340. doi: 10.1038/343336a0. [DOI] [PubMed] [Google Scholar]
  9. Haskill S., Martin G., Van Le L., Morris J., Peace A., Bigler C. F., Jaffe G. J., Hammerberg C., Sporn S. A., Fong S. cDNA cloning of an intracellular form of the human interleukin 1 receptor antagonist associated with epithelium. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3681–3685. doi: 10.1073/pnas.88.9.3681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hattori T., Kubota S., Yutani Y., Fujisawa T., Nakanishi T., Takahashi K., Takigawa M. Change in cellular localization of a rheumatoid arthritis-related antigen (RA-A47) with downregulation upon stimulation by inflammatory cytokines in chondrocytes. J Cell Physiol. 2001 Feb;186(2):268–281. doi: 10.1002/1097-4652(200002)186:2<168::aid-jcp1022>3.0.co;2-m. [DOI] [PubMed] [Google Scholar]
  11. Klareskog L., Forsum U., Malmnäs Tjernlund U. K., Kabelitz D., Wigren A. Appearance of anti-HLA-DR-reactive cells in normal and rheumatoid synovial tissue. Scand J Immunol. 1981 Aug;14(2):183–192. doi: 10.1111/j.1365-3083.1981.tb00198.x. [DOI] [PubMed] [Google Scholar]
  12. Liu J. S., Amaral T. D., Brosnan C. F., Lee S. C. IFNs are critical regulators of IL-1 receptor antagonist and IL-1 expression in human microglia. J Immunol. 1998 Aug 15;161(4):1989–1996. [PubMed] [Google Scholar]
  13. Lotz M., Blanco F. J., von Kempis J., Dudler J., Maier R., Villiger P. M., Geng Y. Cytokine regulation of chondrocyte functions. J Rheumatol Suppl. 1995 Feb;43:104–108. [PubMed] [Google Scholar]
  14. Lotz M., Clark-Lewis I., Ganu V. HIV-1 transactivator protein Tat induces proliferation and TGF beta expression in human articular chondrocytes. J Cell Biol. 1994 Feb;124(3):365–371. doi: 10.1083/jcb.124.3.365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Malyak M., Guthridge J. M., Hance K. R., Dower S. K., Freed J. H., Arend W. P. Characterization of a low molecular weight isoform of IL-1 receptor antagonist. J Immunol. 1998 Aug 15;161(4):1997–2003. [PubMed] [Google Scholar]
  16. Martel-Pelletier J., Alaaeddine N., Pelletier J. P. Cytokines and their role in the pathophysiology of osteoarthritis. Front Biosci. 1999 Oct 15;4:D694–D703. doi: 10.2741/martel. [DOI] [PubMed] [Google Scholar]
  17. Muzio M., Polentarutti N., Facchetti F., Peri G., Doni A., Sironi M., Transidico P., Salmona M., Introna M., Mantovani A. Characterization of type II intracellular IL-1 receptor antagonist (IL-1ra3): a depot IL-1ra. Eur J Immunol. 1999 Mar;29(3):781–788. doi: 10.1002/(SICI)1521-4141(199903)29:03<781::AID-IMMU781>3.0.CO;2-0. [DOI] [PubMed] [Google Scholar]
  18. Muzio M., Polentarutti N., Sironi M., Poli G., De Gioia L., Introna M., Mantovani A., Colotta F. Cloning and characterization of a new isoform of the interleukin 1 receptor antagonist. J Exp Med. 1995 Aug 1;182(2):623–628. doi: 10.1084/jem.182.2.623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nicoletti F., Patti F., DiMarco R., Zaccone P., Nicoletti A., Meroni P., Reggio A. Circulating serum levels of IL-1ra in patients with relapsing remitting multiple sclerosis are normal during remission phases but significantly increased either during exacerbations or in response to IFN-beta treatment. Cytokine. 1996 May;8(5):395–400. doi: 10.1006/cyto.1996.0054. [DOI] [PubMed] [Google Scholar]
  20. Oritani K., Kincade P. W., Zhang C., Tomiyama Y., Matsuzawa Y. Type I interferons and limitin: a comparison of structures, receptors, and functions. Cytokine Growth Factor Rev. 2001 Dec;12(4):337–348. doi: 10.1016/s1359-6101(01)00009-0. [DOI] [PubMed] [Google Scholar]
  21. Palmer Gaby, Guerne Pierre-Andre, Mezin Francoise, Maret Michel, Guicheux Jerome, Goldring Mary B., Gabay Cem. Production of interleukin-1 receptor antagonist by human articular chondrocytes. Arthritis Res. 2002 Apr 8;4(3):226–231. doi: 10.1186/ar411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Palmer Gaby, Talabot-Ayer Dominique, Szalay-Quinodoz Ildiko, Maret Michel, Arend William P., Gabay Cem. Mice transgenic for intracellular interleukin-1 receptor antagonist type 1 are protected from collagen-induced arthritis. Eur J Immunol. 2003 Feb;33(2):434–440. doi: 10.1002/immu.200310018. [DOI] [PubMed] [Google Scholar]
  23. Perini P., Tiberio M., Sivieri S., Facchinetti A., Biasi G., Gallo P. Interleukin-1 receptor antagonist, soluble tumor necrosis factor-alpha receptor type I and II, and soluble E-selectin serum levels in multiple sclerosis patients receiving weekly intramuscular injections of interferon-beta1a. Eur Cytokine Netw. 2000 Mar;11(1):81–86. [PubMed] [Google Scholar]
  24. Sciacca F. L., Canal N., Grimaldi L. M. Induction of IL-1 receptor antagonist by interferon beta: implication for the treatment of multiple sclerosis. J Neurovirol. 2000 May;6 (Suppl 2):S33–S37. [PubMed] [Google Scholar]
  25. Smeets T. J., Dayer J. M., Kraan M. C., Versendaal J., Chicheportiche R., Breedveld F. C., Tak P. P. The effects of interferon-beta treatment of synovial inflammation and expression of metalloproteinases in patients with rheumatoid arthritis. Arthritis Rheum. 2000 Feb;43(2):270–274. doi: 10.1002/1529-0131(200002)43:2<270::AID-ANR5>3.0.CO;2-H. [DOI] [PubMed] [Google Scholar]
  26. Tak P. P., Hart B. A., Kraan M. C., Jonker M., Smeets T. J., Breedveld F. C. The effects of interferon beta treatment on arthritis. Rheumatology (Oxford) 1999 Apr;38(4):362–369. doi: 10.1093/rheumatology/38.4.362. [DOI] [PubMed] [Google Scholar]
  27. Taniguchi T., Takaoka A. A weak signal for strong responses: interferon-alpha/beta revisited. Nat Rev Mol Cell Biol. 2001 May;2(5):378–386. doi: 10.1038/35073080. [DOI] [PubMed] [Google Scholar]
  28. Watson J. M., Lofquist A. K., Rinehart C. A., Olsen J. C., Makarov S. S., Kaufman D. G., Haskill J. S. The intracellular IL-1 receptor antagonist alters IL-1-inducible gene expression without blocking exogenous signaling by IL-1 beta. J Immunol. 1995 Nov 1;155(9):4467–4475. [PubMed] [Google Scholar]
  29. van Holten Judith, Plater-Zyberk Christine, Tak Paul P. Interferon-beta for treatment of rheumatoid arthritis? Arthritis Res. 2002 Sep 18;4(6):346–352. doi: 10.1186/ar598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. van de Loo Fons A. J., van den Berg Wim B. Gene therapy for rheumatoid arthritis. Lessons from animal models, including studies on interleukin-4, interleukin-10, and interleukin-1 receptor antagonist as potential disease modulators. Rheum Dis Clin North Am. 2002 Feb;28(1):127–149. doi: 10.1016/s0889-857x(03)00073-5. [DOI] [PubMed] [Google Scholar]

Articles from Annals of the Rheumatic Diseases are provided here courtesy of BMJ Publishing Group

RESOURCES