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. 1996 Mar;103(3):357–367. doi: 10.1111/j.1365-2249.1996.tb08288.x

Expression of interferon-gamma (IFN-δ), IL-10, IL-12 and transforming growth factor-beta (TGF-β) mRNA in synovial fluid cells from patients in the early and late phases of rheumatoid arthritis (RA)

A BUCHT *,†, P LARSSON †, L WEISBROT , C THORNE ‡, P PISA §, G SMEDEGÅRD †, E C KEYSTONE , A GRÖNBERG †
PMCID: PMC2200369  PMID: 8608632

Abstract

The expression of immunoregulatory cytokines was investigated in freshly isolated synovial fluid mononuclear cells (SFMC) and peripheral blood mononuclear cells (PBMC) from patients with RA, using a quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) assay. IFN-γ, TGF-β, IL-10 and IL-12 (p40) transcripts were detected in SFMC of patients with early disease (<1 year duration) as well as in patients with long standing arthritis (>1 year). The expression of IFN-γ, IL-10 and IL-12 mRNA was increased in SFMC compared with RA PBMC. In addition, the expression was higher in RA SFMC than in PBMC from healthy control individuals. Immunoassay analysis of the secreted IL-12 heterodimer demonstrated increased levels in RA SF compared with levels found in serum from RA patients and control individuals. High levels of TGF-β mRNA were found in SFMC, but a significantly decreased TGF-β/β2-microglobulin (β2-M) ratio was found compared with PBMC from both patients and control individuals. IL-4 mRNA could not be detected, either in SFMC or in PBMC. Cytokine expression in RA PBMC did not differ from control PBMC, with the exception of a decreased TGF-β/β2-M ratio in RA patients with early disease. Our findings of IFN-7 mRNA and IL-12, but undetectable levels of IL-4 mRNA, suggest that the synovitis is characterized by a type 1 immune response. The presence of TGF-β and IL-10 mRNA indicates that immunosuppressive cytokines may also operate in the inflamed joint, although their level of expression may not be sufficient for down-modulation of immune activation.

Keywords: rheumatoid arthritis, interferon-gamma, transforming growth factor-beta, IL-4, IL-10, IL-12

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References

  • 1.Arend WP, Dayer JM. Cytokines and cytokine inhibitors or antagonists in rheumatoid arthritis. Arthritis Rheum. 1990;3:305–15. doi: 10.1002/art.1780330302. [DOI] [PubMed] [Google Scholar]
  • 2.Elliott MJ, Maini RN, Feldmann M, et al. Randomised double-blind comparison of chimeric monoclonal antibody to tumor necrosis factor α (cA2) vs. placebo in rheumatoid arthritis. Lancet. 1994;344:1105–10. doi: 10.1016/s0140-6736(94)90628-9. [DOI] [PubMed] [Google Scholar]
  • 3.Yocum DE, Esparza L, Dubry S, Benjamin JB, Volz R, Scuder P. Characteristics of tumor necrosis factor production in rheumatoid arthritis. Cell Immunol. 1989;122:131–45. doi: 10.1016/0008-8749(89)90154-8. [DOI] [PubMed] [Google Scholar]
  • 4.Katsikis PD, Chu CQ, Brennan FM, Maini RN, Feldmann M. Immunoregulatory role of interleukin 10 in rheumatoid arthritis. J Exp Med. 1994;179:1517–27. doi: 10.1084/jem.179.5.1517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Essner R, Rhoades K, McBride WH, Morton DL, Economou JS. IL4 down-regulates IL1 and TNF gene expression in human monocytes. J Immunol. 1989;142:3857–61. [PubMed] [Google Scholar]
  • 6.Donnelly RP, Fenton MJ, Finbloom DS, Gerrard TL. Differential regulation of IL1 production in human monocytes by IFNγ and IL4. J Immunol. 1990;145:569–75. [PubMed] [Google Scholar]
  • 7.Oswald IP, Wynn TA, Sher A, James SL. Interleukin 10 inhibits macrophage microbicidal activity by blocking the endogenous production of tumor necrosis factor α required as a costimulatory factor for interferon γ-induced activation. Proc Natl Acad Sci USA. 1992;89:8676–80. doi: 10.1073/pnas.89.18.8676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Clerici M, Shearer GM. The Thl-Th2 hypothesis of HIV infection: new insights. Immunol Today. 1994;15:575–81. doi: 10.1016/0167-5699(94)90220-8. [DOI] [PubMed] [Google Scholar]
  • 9.Maggi E, Parronchi P, Manetti R, et al. Reciprocal regulatory effects of IFNγ and IL4 on the in vitro development of human Th1 and Th2 clones. J Immunol. 1992;148:2142–7. [PubMed] [Google Scholar]
  • 10.Tripp CS, Wolf SF, Unanue ER. Interleukin 12 and tumor necrosis factor α are costimulators of interferon γ production by natural killer cells in severe combined immunodeficiency mice with listeriosis and interleukin 10 is a physiologic antagonist. Proc Natl Acad Sci USA. 1993;90:3725–9. doi: 10.1073/pnas.90.8.3725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Dándrea A, Aste-Amezaga M, Valiante NM, Ma X, Kubin M, Trinchieri G. Interleukin 10 (IL10) inhibits human lymphocyte interferon γ-production by suppressing natural killer cell stimulatory factor/IL12 synthesis in accessory cells. J Exp Med. 1993;178:1041–8. doi: 10.1084/jem.178.3.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Fontana A, Constam DB, Frei K, Malipiero U, Pfister HW. Modulation of the immune response by transforming growth factor beta. Int Arch Allergy. 1992;99:1–7. doi: 10.1159/000236328. [DOI] [PubMed] [Google Scholar]
  • 13.Thorbecke GJ, Shah R, Leu CH, Kuruvilla AP, Hardison AM, Palladino MA. Involvement of endogenous tumor necrosis factor α and transforming growth factor β during induction of collagen type II arthritis in mice. Proc Natl Acad Sci USA. 1992;89:7375–9. doi: 10.1073/pnas.89.16.7375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Fava RA, Olsen NJ, Postlethwaite AE, et al. Transforming growth factor β1 (TGFβ1) induced neutrophil recruitment to synovial tissues: implication for TGFβ-driven synovial inflammation and hyperplasia. J Exp Med. 1991;173:1121–32. doi: 10.1084/jem.173.5.1121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15.Kahle P, Saal G, Schaudt K, Zacher J, Fritz P, Pawelec G. Determination of cytokines in synovial fluids: correlation with diagnosis and histomorphological characteristics of synovial tissue. Ann Rheum Dis. 1992;51:731–4. doi: 10.1136/ard.51.6.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Firestein GS, Alvaro-Garcia JM, Maki R. Quantitative analysis of cytokine gene expression in rheumatoid arthritis. J Immunol. 1990;144:3347–53. [PubMed] [Google Scholar]
  • 17.Farahat MN, Yanni G, Poston R, Panayi GS. Cytokine expression in synovial membranes of patients with rheumatoid arthritis and osteoarthritis. Ann Rheum Dis. 1993;52:870–5. doi: 10.1136/ard.52.12.870. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18.Wood NC, Symons JA, Dickens E, Duff GW. In situ hybridization of IL6 in rheumatoid arthritis. Clin Exp Immunol. 1992;87:183–9. doi: 10.1111/j.1365-2249.1992.tb02972.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Field M, Chu D, Feldmann M, Maini RN. Interleukin-6 localisation in the synovial membrane in rheumatoid arthritis. Rheumatol Int. 1991;11:45–50. doi: 10.1007/BF00291144. [DOI] [PubMed] [Google Scholar]
  • 20.Houssiau FA, Devogelaer JP, Van Damme J, de Deuxchaisnes CN, van Snick J. Interleukin-6 in synovial fluid and serum of patients with rheumatoid arthritis and other inflammatory arthritides. Arthritis Rheum. 1988;31:784–8. doi: 10.1002/art.1780310614. [DOI] [PubMed] [Google Scholar]
  • 21.Koch AE, Kunkel SL, Burrows JC, Evanoff HL, Haines GK, Pope RM, Streiter RM. Synovial tissue macrophage as a source of the chemotactic cytokine IL8. J Immunol. 1991;147:2187–95. [PubMed] [Google Scholar]
  • 22.Hopkins SJ, Meager A. Cytokines in synovial fluid: II. The presence of tumor necrosis factor and interferon. Clin Exp Immunol. 1988;73:88–92. [PMC free article] [PubMed] [Google Scholar]
  • 23.Xu W, Firestein GS, Taetle R, Kaushansky K, Zvaifler NJ. Cytokines in chronic inflammatory arthritis. J Clin Invest. 1989;83:876–82. doi: 10.1172/JCI113971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Fava R, Olsen N, Keski-Oja J, Moses H, Pincus T. Active and latent forms of transforming growth factor β activity in synovial effusions. J Exp Med. 1989;169:291–6. doi: 10.1084/jem.169.1.291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25.Miossec P, Naviliat M, DÁngeac AD, Sany J, Banchereau J. Low levels of interleukin-4 and high levels of transforming growth factor β in rheumatoid synovitis. Arthritis Rheum. 1990;33:1180–7. doi: 10.1002/art.1780330819. [DOI] [PubMed] [Google Scholar]
  • 26.Lafyatis R, Thompson NL, Remmers EF, et al. Transforming growth factor-β production by synovial tissues from rheumatoid arthritis patients and streptococcal cell wall arthritic rats. J Immunol. 1989;143:1142–8. [PubMed] [Google Scholar]
  • 27.Saxne T, Palladino MA, Heinegárd D, Talal N, Wollheim F. Detection of tumor necrosis factor α but not tumor necrosis factor β in rheumatoid arthritis synovial fluid and serum. Arthritis Rheum. 1988;31:1041–5. doi: 10.1002/art.1780310816. [DOI] [PubMed] [Google Scholar]
  • 28.Simon AK, Seipelt E, Sieper J. Divergent T-cell cytokine patterns in inflammatory arthritis. Proc Natl Acad Sci USA. 1994;91:8562–6. doi: 10.1073/pnas.91.18.8562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29.Llorente L, Richaud-Patin Y, Fior R, et al. In vivo production of interleukin-10 by non-T cells in rheumatoid arthritis, Sjögren's syndrome, and systemic lupus erythematosus. Arthritis Rheum. 1994;37:1647–55. doi: 10.1002/art.1780371114. [DOI] [PubMed] [Google Scholar]
  • 30.Potocnik AJ, Kinne R, Menninger H, Zacher J, Emmrich F, Kroczek RA. Expression of activation antigens on T cells in rheumatoid arthritis patients. Scand J Immunol. 1990;31:213–24. doi: 10.1111/j.1365-3083.1990.tb02762.x. [DOI] [PubMed] [Google Scholar]
  • 31.Iannone F, Corrigall VM, Kingsley GH, Panayi G. Evidence for the continuous recruitment and activation of T cells into the joints of patients with rheumatoid arthritis. Eur J Immunol. 1994;24:2706–13. doi: 10.1002/eji.1830241120. [DOI] [PubMed] [Google Scholar]
  • 32.Firestein GS, Zvaifler NJ. Peripheral blood and synovial fluid monocyte activation in inflammatory arthritis. II. Low levels of synovial fluid and synovial tissue interferon suggest that gamma-interferon is not the primary macrophage activating factor. Arthritis Rheum. 1987;30:864–71. doi: 10.1002/art.1780300804. [DOI] [PubMed] [Google Scholar]
  • 33.Yamagata N, Kobayashi K, Kasama T, et al. Multiple cytokine activities and loss of inteleukin 2 inhibitor in synovial fluids of patients with rheumatoid arthritis. J Rheumatol. 1988;15:1623–7. [PubMed] [Google Scholar]
  • 34.Buchan G, Barrett K, Fujita T, Taniguchi T, Maini R, Feldmann M. Detection of activated T cell products in the rheumatoid joint using cDNA probes to interleukin-2 (IL2), IL2 receptor and IFNγ. Clin Exp Immunol. 1988;71:295–301. [PMC free article] [PubMed] [Google Scholar]
  • 35.Chen E, Keystone EC, Fish EN. Restricted cytokine expression in rheumatoid arthritis. Arthritis Rheum. 1993;36:901–10. doi: 10.1002/art.1780360706. [DOI] [PubMed] [Google Scholar]
  • 36.Arnett FC, Edworthy SM, Bloch DA, et al. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum. 1988;31:315–24. doi: 10.1002/art.1780310302. [DOI] [PubMed] [Google Scholar]
  • 37.Chomczynski P, Sacchi N. Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987;162:156–9. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  • 38.Suggs SV, Wallace B, Hirose T, Kawashima EH, Itakura K. Use of synthetic oligonucleotides as hybridization probes: isolation of cloned cDNA sequences for human β2-microglobulin. Proc Natl Acad Sci USA. 1981;78:6613–7. doi: 10.1073/pnas.78.11.6613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 39.Choi Y, Kotzin B, Herron L, Callahan J, Marrack P, Kappler J. Interaction of Staphylococcus aureus toxin ‘superantigens’ with human T cells. Proc Natl Acad Sci USA. 1989;86:8941–5. doi: 10.1073/pnas.86.22.8941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40.Gray PW, Goeddel DV. Structure of the human immune interferon gene. Nature. 1982;298:859–63. doi: 10.1038/298859a0. [DOI] [PubMed] [Google Scholar]
  • 41.Derynck R, Jarrett JA, Chen EY, et al. Human transforming growth factor-β complementary DNA sequence and expression in normal and transformed cells. Nature. 1985;316:701–5. doi: 10.1038/316701a0. [DOI] [PubMed] [Google Scholar]
  • 42.Yokota T, Otsuka T, Mosman T, et al. Isolation and characterization of a human interleukin cDNA clone, homologous to mouse B-cell stimulatory factor 1, that express B-cell- and T-cell-stimulating activities. Proc Natl Acad Sci USA. 1986;83:5894–8. doi: 10.1073/pnas.83.16.5894. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 43.Vieira P, de Waal-Malefyt R, Dang MN, et al. Isolation and expression of human cytokine synthesis inhibitory factor cDNA clones: homology to Epstein-Barr virus open reading frame BCRF1. Proc Natl Acad Sci USA. 1991;88:1172–6. doi: 10.1073/pnas.88.4.1172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 44.Wolf SF, Temple PA, Kobayashi M, et al. Cloning of cDNA for natural killer cell stimulatory factor, a heterodimeric cytokine with multiple biologic effects on T and natural killer cells. J Immunol. 1991;146:3074–81. [PubMed] [Google Scholar]
  • 45.Bucht A, Söderström K, Esin S, et al. Analysis of γδ V region usage in normal and diseased human intestinal biopsies and peripheral blood by polymerase chain reaction (PCR) and flow cytometry. Clin Exp Immunol. 1995;99:57–64. doi: 10.1111/j.1365-2249.1995.tb03472.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 46.Réme T, Portier M, Frayssinoux F, Combe B, Miossec P, Favier F, Sany J. T cell receptor expression and activation of synovial lymphocyte subsets in patients with rheumatoid arthritis. Arthritis Rheum. 1990;33:485–92. doi: 10.1002/art.1780330404. [DOI] [PubMed] [Google Scholar]
  • 47.Tang MLK, Varigos G, Kemp AS. Reduced interferon-gamma (IFN-γ) secretion with increased IFN-γ mRNA expression in atopic dermatitis: evidence for a post-transcriptional defect. Clin Exp Immunol. 1994;97:483–90. doi: 10.1111/j.1365-2249.1994.tb06114.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 48.Oswald IP, Gazzinelli RT, Sher A, James SL. IL10 synergizes with IL4 and transforming growth factor-β to inhibit macrophage cytotoxic activity. J Immunol. 1992;148:3578–82. [PubMed] [Google Scholar]
  • 49.Abeshira-Amar O, Gibert M, Joliy M, Théze J, Jankovic D. IL4 plays a dominant role in the differential development of Th0 into Th1 and Th2 cells. J Immunol. 1992;148:3820–9. [PubMed] [Google Scholar]
  • 50.Schmitt E, Hoehn P, Huels C, Goedert S, Palm N, Rüde E, Germann T. T helper type 1 development requires the coordinate action of interleukin-12 and interferon-γ and is inhibited by transforming growth factor-β. Eur J Immunol. 1994;24:793–8. doi: 10.1002/eji.1830240403. [DOI] [PubMed] [Google Scholar]
  • 51.Kennedy MK, Picha KS, Shanebeck KD, Anderson DM, Grabstein KH. Interleukin-12 regulates the proliferation of Th1, but not Th2 or Th0, clones. Eur J Immunol. 1994;24:2271–8. doi: 10.1002/eji.1830241002. [DOI] [PubMed] [Google Scholar]
  • 52.Ling P, Gately MK, Gubler U, et al. Human IL-12 p40 homodimer binds to the IL-12 receptor but does not mediate biologic activity. J Immunol. 1995;154:116–27. [PubMed] [Google Scholar]
  • 53.Wahl SM, Allen JB, Wong HL, Dougherthy SF, Ellingsworth LR. Antagonistic and agonistic effects of transforming growth factor-β and IL-1 in rheumatoid synovium. J Immunol. 1990;145:2514–9. [PubMed] [Google Scholar]
  • 54.Olsson T. Role of cytokines in multiple sclerosis and experimental autoimmune encephalomyelitis. Eur J Neurol. 1994;1:7–19. doi: 10.1111/j.1468-1331.1994.tb00045.x. [DOI] [PubMed] [Google Scholar]
  • 55.Khoury SJ, Hancock WW, Weiner H. Oral tolerance to myelic basic protein and natural recovery from experimental autoimmune encephalomyelitis are associated with downregulation of inflammatory cytokines and differential upregulation of transforming growth factor β, interleukin 4 and prostaglandin E expression in the brain. J Exp Med. 1992;176:1355–64. doi: 10.1084/jem.176.5.1355. [DOI] [PMC free article] [PubMed] [Google Scholar]

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