Abstract
The destruction of cartilage is an important characteristic of rheumatoid arthritis (RA). Immune complexes (IC) are usually found in high amounts in RA synovial fluids (SF) and in the superficial layers of RA cartilage. The objective of this study was to investigate if IC have a direct influence on proliferation, survival and production of nitric oxide (NO) of cytokine-activated chondrocytes. Primary bovine chondrocytes were incubated with cytokines (huIL-1α, bovIFN-γ, huTNF-α) and IC containing precipitates of peripheral blood (PB) and/or synovial fluid (SF) of 14 RA patients, 5 osteoarthritis (OA) patients and 10 healthy age and sex-matched controls. After 48 h, chondrocyte viability, proliferation, apoptosis, NO production and oxygen radical levels were measured. Staining with May-Grünwald-Giemsa after incubation with IC of RA PB and SF, showed apoptotic chondrocytes with condensation of the nuclei. The proliferation rates of cytokine-activated chondrocytes, incubated with sera and SF IC of RA patients were significantly decreased compared to chondrocytes, incubated with sera and SF IC of OA patients and compared to sera of controls. Quantitative evaluation of apoptotic cells by annexin-V/propidium iodide and TUNEL assays revealed a significant increase after incubation with sera and SF IC of RA patients, compared to control sera and OAs sera and SF. In all TUNEL positive samples, active-caspase-3-positive cells were found. There was a significant increase of chondrocyte NO production, after incubation with SF IC of RA patients, compared to OA SF. These results support the hypothesis that IC, present in serum and SF of RA patients, have a profound influence on chondrocyte growth, NO production and apoptosis, contributing to cartilage destruction in RA.
Similar content being viewed by others
References
Choy EH, Panayi GS (2001) Cytokine pathways and joint inflammation in rheumatoid arthritis. N Engl J Med 344(12):907–916
Salmon JE, Pricop L (2001) Human receptors for immunoglobulin G: key elements in the pathogenesis of rheumatic disease. Arthritis Rheum 44(4):739–750
Bonomo L, Tursi A, Trizio D, Gillardi U, Dammacco F (1970) Immune complexes in rheumatoid synovitis: a mixed staining immunofluorescence study. Immunology 18(4):557–563
Mellbye OJ, Natvig JB (1971) Evidence for immune complexes containing antibody to the pepsin site of IgG in rheumatoid synovial fluids. Clin Exp Immunol 8(6):889–899
De Clerck LS, Struyf NJ, Bridts CH, Franckx L, Van Offel JF, Empsten FA et al (1989) Humoral immunity and composition of immune complexes in patients with rheumatoid arthritis, with special reference to IgE-containing immune complexes. Clin Exp Rheumatol 7(5):485–492
Cooke D, Saura R, Uno K, Scudamore A (1991) Interactions of immunoglobulins and chondrocytes. J Rheumatol Suppl 27:114–116
Cooke TD, Hurd ER, Ziff M, Jasin HE (1975) The pathogenesis of chronic inflammation in experimental antigen induced arthritis. Preferential localization of antigen-antibody complexes to collageneous tissues. J Exp Med 135:323–338
Alomari WR, Archer JR, Brocklehurst R, Currey HL (1983) Binding of immunoglobulins and immune complexes to cartilage derived extracts. Clin Exp Immunol 54(3):716–722
De Clerck LS, Struyf NJ, Bridts CH, van Marck EA, Breedveld FC, Devries E et al (1992) Experimental arthritis in rats induced by intra-articular injection of IgE aggregates: evidence for arthritogenic role of complexed IgE. Ann Rheum Dis 51(2):210–213
Dishon T (1971) Experimental arthritis in the rabbit induced by intra-articular injection of immune complexes. J Dent Res 50(3):651
Dumonde DC, Glynn LE (1962) The production of arthritis in rabbits by an immunological reaction to fibrin. Br J Exp Pathol 43:373–383
Johnson KJ, Ward PA (1982) Biology of disease. Newer concepts in the pathogenesis of immune complex-induced tissue injury. Lab Invest 47(3):218–226
Sumi M, Maeda M, Cooke TD (1986) Deleterious interactions of immune complexes with tibial cartilage of antigen-induced arthritic rabbits. II: chondrocyte degradation Clin Orthop 212:260–274
Takagi T, Jasin HE (1992) Interactions of synovial fluid immunoglobulins with chondrocytes. Arthritis Rheum 35:1502–1509
Saura R, Uno K, Satsuma S, Kurz EU, Scudamore RA, Cooke TD (1993) Mechanisms of cartilage degradation in inflammatory arthritis: interaction between chondrocytes and immunoglobulin G. J Rheumatol 20(2):336–343
Verbruggen A, De Clerck LS, Bridts CH, Breedveld FC, Stevens WJ (2000) Influence of blood and synovial fluid immune complexes of patients with rheumatoid arthritis on production of nitric oxide and growth and viability of chondrocytes. J Rheumatol 27(1):35–40
Arnett FC, Edworthy SM, Bloch DA, McShane DJ, Fries JF, Cooper NS et al (1988) The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum 31(3):315–324
Altman E, Asch D, Bloch G, Bole D, Borenstein K, Brandt et al (1986) The American College of Rheumatology criteria for the classification and reporting of osteoarthritis of the knee. Arthritis Rheum 29:1039–1049
Stevens WJ, Bridts C (1981) A method for rapid determination of IgG containing circulating immune complexes using polyethyleneglycol and radioactively labeled protein A. Immunol Lett 3(1):1–4
Clancy RM, Abramson SB, Kohne C, Rediske J (1997) Nitric oxide attenuates cellular hexose monophosphate shunt response to oxidants in articular chondrocytes and acts to promote oxidant injury. J Cell Physiol 172(2):183–191
Schuerwegh AJ, Dombrecht EJ, Stevens WJ, Van Offel JF, Bridts CH, De Clerck LS (2003) Influence of pro-inflammatory (IL-1α, IL-6, TNF-α, IFN-γ) and anti-inflammatory (IL-4) cytokines on chondrocyte function. Osteoarthr Cartil 11:681–687
Kockx MM, Muhring J, Knaapen MW, de Meyer GR (1998) RNA synthesis and splicing interferes with DNA in situ end labeling techniques used to detect apoptosis. Am J Pathol 152(4):885–888
Tiku ML, Liesch JB, Robertson FM (1990) Production of hydrogen peroxide by rabbit articular chondrocytes. Enhancement by cytokines. J Immunol 145(2):690–696
Westacott CI, Whicher JT, Barnes IC, Thompson D, Swan AJ, Dieppe PA (1990) Synovial fluid concentration of five different cytokines in rheumatic diseases. Ann Rheum Dis 49(9):676–681
Schlaak JF, Pfers I, Meyer Zum Buschenfelde KH, Marker-Hermann E (1996) Different cytokine profiles in the synovial fluid of patients with osteoarthritis, rheumatoid arthritis and seronegative spondylarthropathies. Clin Exp Rheumatol 14(2):155–162
Henrotin YE, Zheng SX, Labasse AH, Deby GP, Crielaard JM, Reginster JY (2000) Modulation of human chondrocyte metabolism by recombinant human interferon. Osteoarthr Cartil 8(6):474–482
Oyanagui Y (1994) Nitric oxide and superoxide radical are involved in both initiation and development of adjuvant arthritis in rats. Life Sci 54(17):L285–L289
Stadler J, Stefanovic-Racic M, Billiar TR, Curran RD, McIntyre LA, Georgescu HI et al (1991) Articular chondrocytes synthesize nitric oxide in response to cytokines and lipopolysaccharide. J Immunol 147(11):3915–3920
Nabbe KC, Boross P, Holthuysen AE, Sloetjes AW, Kolls JK, Verbeek S, van Lent PL, van Den Berg WB (2005) Joint inflammation and chondrocyte death become independent of Fcgamma receptor type III by local overexpression of interferon-gamma during immune complex-mediatedarthritis. Arthritis Rheum 52(3):967–974
Nabbe KC, van Lent PL, Holthuysen AE, Sloetjes AW, Koch AE, Radstake TR, van den Berg WB (2005) Local IL-13 gene transfer prior to immune-complex arthritis inhibits chondrocyte death and MMP-mediated cartilage matrix degradation despite enhanced joint inflammation. Arthritis Res Ther 7(2):R392–R401. Epub
Nydegger UE, Zubler RH, Gabay R, Joliat G, Karagevrekis CH, Lambert PH et al (1977) Circulating complement breakdown products in patients with rheumatoid arthritis. Correlation between plasma C3d, circulating immune complexes, and clinical activity. J Clin Invest 59(5):862–868
Levo Y, Fischel R, Ehrenfeld M (1981) Circulating immune complexes in patients with rheumatoid arthritis: correlation with disease activity. J Rheumatol 8(5):851–855
Lee DA, Salih V, Stockton EF, Stanton JS, Bentley G (1997) Effect of normal synovial fluid on the metabolism of articular chondrocytes in vitro. Clin Orthop 342:228–238
Jarvis JN, Wang W, Moore HT, Zhang L, Xu C (1997) In vitro induction of proinflammatory cytokine secretion by juvenile rheumatoid arthritis synovial fluid immune complexes. Arthritis Rheum 11:2039–2046
Tighe H, Warnatz K, Brinson D, Corr M, Weigle WO, Baird SM et al (1997) Peripheral deletion of rheumatoid factor B cells after abortive activation by IgG. Proc Natl Acad Sci USA 94(2):646–651
Aceituno E, Castanon S, Jimenez C, Subira D, De Gorgolas M, Fernandez-Guerrero M et al (1997) Circulating immune complexes from HIV-1+ patients induces apoptosis on normal lymphocytes. Immunology 92(3):317–320
Gamberale R, Giordano M, Trevani AS, Andonegui G, Geffner JR (1998) Modulation of human neutrophil apoptosis by immune complexes. J Immunol 161(7):3666–3674
Ortaldo JR, Mason AT, O’Shea JJ (1995) Receptor-induced death in human natural killer cells: involvement of CD16. J Exp Med 181(1):339–344
Crompton M (1999) The mitochondrial permeability transition pore and its role in cell death. Biochem J 341:233–249
Tomita M, Sato EF, Nishikawa M, Yamano T, Inoue M (2001) Nitric oxide regulates mitochondrial respiration and function of chondrocytes. Arthritis Rheum 44:96–104
Paul A, Wilson S, Belham CM, Robinson CJ, Scott PH, Gould GW, Plevin R (1997) Stress-activated protein kinases : activation, regulation and function. Cell Signal 9:403–410
Stefanovic-Racic M, Evans CH (1993) Nitric oxide and arthritis. Arthritis Rheum 36:1036
Fukuda K, Kumano F, Takayama M, Saito M, Otani K, Tanaka S (1995) Zonal differences in nitric oxide synthesis by bovine chondrocytes exposed to interleukin-1. Inflamm Res 44:434
Hayashi T, Abe E, Yamate T, Taguchi Y, Jasin HE (1997) Nitric oxide production by superficial and deep chondrocytes. Arthritis Rheum 40:26
Blanco J, Ochs R, Schwartz H, Lotz M (1995) Chondrocyte apoptosis induced by nitric oxide. Am J Pathol 146:75–85
Acknowledgments
We thank the staff of the policlinic of Immunology-Allergology-Rheumatology of the University Hospital of Antwerp, Dr. L. Goethals and Dr. L. Francx for their support in SF and blood sampling. Further, we want to thank Karin Van Cotthem and Christel Mertens for their expert technical assistance. This study was supported by a grant from the Fund of Scientific Investigation (FWO)-Flanders-Belgium.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Schuerwegh, A.J., Dombrecht, E.J., Stevens, W.J. et al. Synovial fluid and peripheral blood immune complexes of patients with rheumatoid arthritis induce apoptosis in cytokine-activated chondrocytes. Rheumatol Int 27, 901–909 (2007). https://doi.org/10.1007/s00296-007-0336-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00296-007-0336-3