Skip to main content
SpringerLink
Log in

Immunohistochemical demonstration of collagenase and tissue inhibitor of metalloproteinases (TIMP) in synovial lining cells of rheumatoid synovium

  • Published:
Virchows Archiv B

Summary

Degradation of fibrillar collagens is a central process in joint destruction in rheumatoid arthritis. Collagenase responsible for the collagenolysis has been immunolocalized on the extracellular matrix components at the cartilage/pannus junction in the rheumatoid joint, but very little is known about cellular source of the proteinase. In this paper monospecific antibodies against collagenase and tissue inhibitor of metalloproteinases (TIMP) were applied to rheumatoid and normal synovium to identify cells synthesizing and secreting the enzyme and its inhibitor. By treating the specimens with the monovalent ionophore, monensin, both collagenase and TIMP could be immunolocalized in hyperplastic synovial lining cells in rheumatoid synovium, but not in the cells of normal synovium. Dual immunolocalization studies demonstrated that the majority of the lining cells (approximately 64%) produce both collagenase and TIMP, while approximately 3% of the cells were positive only for collagenase, and 11 % only for TIMP. Neither collagenase nor TIMP was immunolocalized on the extracellular matrix components in the synovia examined. These data suggest that synovial lining cells in rheumatoid arthritis secrete both collagenase and TIMP into the joint cavity. The role of collagenase in joint destruction in rheumatoid arthritis is discussed with reference to the regulation of the activity by TIMP.

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

  • Abe S, Shinmei M, Nagai Y (1973) Synovial collagenase and joint diseases: The significancy of latent collagenase with special reference to rheumatoid arthritis. J Biochem 73:1007–1011

    PubMed  CAS  Google Scholar 

  • Arnett FC, Edworthy SM, Block DA, McShane DJ, Fries JF, Cooper NS, Healey LA, Kaplan SR, Liang MH, Luthra HS, Medsger TA Jr, Mitchell DM, Neustadt DH, Pinals RS, Schlaller JG, Sharp JT, Wilder RL, Hunder GG (1988) The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum 31:315–324

    Article  PubMed  CAS  Google Scholar 

  • Barland P, Novikoff AB, Hamerman D (1962) Electron microscopy of the human synovial membrane. J Cell Biol 14:207–220

    Article  PubMed  CAS  Google Scholar 

  • Cawston TE, Barrett AJ (1979) A rapid and reproducible assay for collagenase using [1-14C] acetylated collagen. Anal Biochem 99:340–345

    Article  PubMed  CAS  Google Scholar 

  • Cawston TE, Galloway WA, Mercer E, Murphy G, Reynolds JJ (1981) Purification of rabbit bone inhibitor of collagenase. Biochem J 195:159–165

    PubMed  CAS  Google Scholar 

  • Cawston TE, Mercer E, De Silva M, Halzleman BL (1984) Metalloproteinases and collagenase inhibitors in rheumatoid synovial fluid. Arthritis Rheum 27:285–290

    Article  PubMed  CAS  Google Scholar 

  • Childers JW, Hernandez AD, Kim JH, Stricklin GP (1987) Immunolocalization of collagenase inhibitor in normal skin and basal cell carcinoma. J Am Acad Dermatol 17:1025–1032

    Article  PubMed  CAS  Google Scholar 

  • Chin JR, Murphy G, Werb Z (1985) Stromelysin, a connective tissue-degrading metalloendopeptidase secreted by stimulated rabbit synovial fibroblasts in parallel with collagenase. J Biol Chem 260:12367–12376

    PubMed  CAS  Google Scholar 

  • Chowcat NL, Savage FJ, Hembry RM, Boulos PB (1988) Role of collagenase in colonic anastomoses: A reappraisal. Br J Surg 75:330–334

    Article  PubMed  CAS  Google Scholar 

  • Dayer J-M, Krane SM, Russell GG, Robinson DR (1976) Production of collagenase and prostaglandins by isolated adherent rheumatoid synovial cells. Proc Natl Acad Sci USA 73:945–949

    Article  PubMed  CAS  Google Scholar 

  • Dayer J-M, Beutler B, Cerami A (1985) Cachectin/tumor necrosis factor stimulates collagenase and prostaglandin E2 production by human synovial cells and dermal fibroblasts. J Exp Med 162:2163–2168

    Article  PubMed  CAS  Google Scholar 

  • Dean DD, Martel-Pelletier J, Pelletier J-P, Howell DS, Woessner JF Jr (1989) Evidence for metalloproteinase and metalloproteinase inhibitor imbalance in human osteoarthritic cartilage. J Clin Invest 84:678–685

    Article  PubMed  CAS  Google Scholar 

  • Evanson JM, Jeffrey JJ, Krane SM (1968) Studies on collagenase from rheumatoid synovium in tissue culture. J Clin Invest 47:2639–2651

    PubMed  CAS  Google Scholar 

  • Galloway WA, Murphy G, Sandy JD, Gavrilovic J, Cawston TE, Reynolds JJ (1983) Purification and characterization of a rabbit bone metalloproteinase that degrades proteoglycan and other connective-tissue components. Biochem J 209:741–752

    PubMed  CAS  Google Scholar 

  • Graabaek PM (1982) Ultrastructural evidence for two distinct types of synoviocytes in rat synovial membrane. J Ultrastruct Res 78:321–339

    Article  PubMed  CAS  Google Scholar 

  • Harris ED Jr, DiBona DR, Krane SM (1969) Collagenases in human synovial fluid. J Clin Invest 48:2104–2113

    Article  PubMed  CAS  Google Scholar 

  • Harris ED Jr, Welgus HG, Krane SM (1984) Regulation of the mammallian collagenases. Collagen Rel Res 4:493–512

    CAS  Google Scholar 

  • Hembry RM, Murphy G, Reynolds JJ (1985) Immunolocalization of tissue inhibitor of metalloproteinases (TIMP) in human cells: Characterization and use of a specific antiserum. J Cell Sci 73:105–119

    PubMed  CAS  Google Scholar 

  • Hembry RM, Ehrlich HP (1986 a) Immunolocalization of collagenase and tissue inhibitor of metalloproteinases (TIMP) in hypertrophic scar tissue. Br J Dermatol 115:409–420

    Article  PubMed  CAS  Google Scholar 

  • Hembry RM, Murphy G, Cawston TE, Dingle JT, Reynolds JJ (1986b) Characterization of a specific antiserum for mammalian collagenase from several species: Immunolocalization of collagenase in rabbit chondrocytes and uterus. J Cell Sci 81:105–123

    PubMed  CAS  Google Scholar 

  • Hoshino T, Kishi J, Kawai T, Kobayashi K, Hayakawa T (1986) Immunoelectron microscopy localization of collagenase inhibitor in bovine dentin. Collagen Rel Res 6:303–312

    CAS  Google Scholar 

  • Ito A, Goshowaki H, Sato T, Mori Y, Yamashita K, Hayakawa T, Nagase H (1988a) Human recombinant interleukin-1 α-mediated stimulation of procollagenase production and suppression of biosynthesis of tissue inhibitor of metalloproteinases in rabbit uterine cervical fibroblasts. FEBS Lett 234:326–330

    Article  PubMed  CAS  Google Scholar 

  • Ito A, Nagase H (1988b) Evidence that human rheumatoid synovial matrix metalloproteinase 3 is an endogenous activator of procollagenase. Arch Biochem Biophys 267:211–216

    Article  PubMed  CAS  Google Scholar 

  • Kishi J, Kawai T, Hayakawa T (1985) Immunohistochemical localization of collagenase inhibitor in bovine dental pulp, pulp cells in monolayer culture, and in some oral connective tissues. Archs Oral Biol 30:571–576

    Article  CAS  Google Scholar 

  • Kodama S, Kishi J, Obata K, Iwata K, Hayakawa T (1987) Monoclonal antibodies to bovine collagenase inhibitor. Collagen Rel Res 7:341–350

    CAS  Google Scholar 

  • Kodama S, Yamashita K, Kishi J, Iwata K, Hayakawa T (1989) A sandwich enzyme immunoassay for collagenase inhibitor using monoclonal antibodies. Matrix 9:1–6

    PubMed  CAS  Google Scholar 

  • Linck G, Porte A (1981) Cytophysiology of the synovial membrane: Distinction of two cell types of the intima revealed by their reaction with horseradish peroxidase and iron saccharate in the mouse. Biol Cell 42:147–152

    Google Scholar 

  • Mainardi CL, Seyer JM, Kang AH (1980) Type-specific collagenolysis: A type V collagen-degrading enzyme from macrophages. Biochem Biophys Res Commun 9:1108–1115

    Google Scholar 

  • McLean IW, Nakane PK (1974) Periodate-lysine-paraformaldehyde fixative: A new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083

    PubMed  CAS  Google Scholar 

  • Mizel SB, Dayer J-M, Krane SM, Merenhagen SE (1981) Stimulation of rheumatoid synovial cell collagenase and prostaglandin production by partially purified lymphocyte-activating factor (interleukin 1). Proc Natl Acad Sci USA 78:2474–2477

    Article  PubMed  CAS  Google Scholar 

  • Morré DJ, Morré DM, Mollenhauser HH, Reutter W (1987) Golgi apparatus cisternae of monensin-treated cells accumulate in the cytoplasm of liver slices. Eur J Cell Biol 43:235–242

    PubMed  Google Scholar 

  • Murphy G, Reynolds JJ, Werb Z (1985) Biosynthesis of tissue inhibitor of metalloproteinases by human fibroblasts in culture: Stimulation by 12-0-tetradecanoylphorbol 13-acetate and interleukin 1 in parallel with collagenase. J Biol Chem 260:3079–3083

    PubMed  CAS  Google Scholar 

  • Murphy G, Hembry RM, Reynolds JJ (1986) Characterization of a specific antiserum to rabbit stromelysin and demonstration of the synthesis of collagenase and stromelysin by stimulated rabbit articular chondrocytes. Collagen Rel Res 6:351–364

    CAS  Google Scholar 

  • Murphy G, Cockett MI, Stephens PE, Smith BJ, Dockerty AJP (1987) Stromelysin is an activator of procollagenase. A study with natural and recombinant enzymes. Biochem J 248:265–268

    PubMed  CAS  Google Scholar 

  • Nagase H, Jackson RC, Brinckerhoff CE, Vater CA, Harris ED Jr (1981) A precursor form of latent collagenase produced in a cell-free system with mRNA from rabbit synovial cells. J Biol Chem 256:11951–11954

    PubMed  CAS  Google Scholar 

  • Nagase H, Brinckerhoff CE, Vater CA, Harris ED Jr (1983) Biosynthesis and secretion of procollagenase by rabbit synovial fibroblasts. Inhibition of procollagenase secretion by monensin and evidence for glycosylation of procollagenase. Biochem J 214:281–288

    PubMed  CAS  Google Scholar 

  • Nolan JC, Ridge SC, Oronsky AL, Kerwar SS (1980) Purification and properties of a collagenase inhibitor from cultures of bovine aorta. Atherosclerosis 35:93–102

    Article  PubMed  CAS  Google Scholar 

  • Okada Y, Nakanishi I, Kajikawa K (1981) Ultrastructure of the mouse synovial membrane: Development and organization of the extracellular matrix. Arthritis Rheum 24:835–843

    Article  PubMed  CAS  Google Scholar 

  • Okada Y, Nakanishi I, Munehiro C, Umeda S, Ichizen H, Masuda S (1984) The presence of siderosomes in synovioblasts (B-cells) of chronic spontaneous hemarthrosis: Histologic, electron microscopic, and roentgenographic electron microanalysis studies of three cases. Arch Pathol Lab Med 108:968–972

    PubMed  CAS  Google Scholar 

  • Okada Y, Nagase H, Harris ED Jr (1986) A metalloproteinase from human rheumatoid synovial fibroblasts that digests connective tissue matrix components. Purification and characterization. J Biol Chem 261:14245–14255

    PubMed  CAS  Google Scholar 

  • Okada Y, Watanabe S, Nakanishi I, Kishi J, Hayakawa T, Watorek W, Travis J, Nagase H (1988) Inactivation of tissue inhibitor of metalloproteinases by neutrophil elastase and other serine proteinases. FEBS Lett 229:157–160

    Article  PubMed  CAS  Google Scholar 

  • Okada Y, Konomi H, Yada T, Kimata K, Nagase H (1989 a) Degradation of type IX collagen by matrix metalloproteinase 3 (stromelysin) from human rheumatoid synovial cells. FEBS Lett 244:473–476

    Article  PubMed  CAS  Google Scholar 

  • Okada Y, Takeuchi N, Tomita K, Nakanishi I, Nagase H (1989 b) Immunolocalization of matrix metalloproteinase 3 (stromelysin) in rheumatoid synovioblasts (B-cells): Correlation to rheumatoid arthritis. Ann Rheum Dis 48:645–653

    PubMed  CAS  Google Scholar 

  • Ouchterlony O, Nilsson LA (1973) Immunodiffusion and immunoelectrophoresis. In: Weir DM (ed) Handbook of experimental immunology. Second edition. Blackwell Scientific Publication, Oxford, pp 19.1–19.39

    Google Scholar 

  • Postlethwaite AE, Raghow R, Stricklin GP, Poppleton H, Seyer JM, Kang AH (1988) Modulation of fibroblast functions by interleukin 1: Increased steady-state accumulation of type I procollagen messenger RNAs and stimulation of other functions but not chemotaxis by human recombinant interleukin 1 α andβ. J Cell Biol 106:311–318

    Article  PubMed  CAS  Google Scholar 

  • Stricklin GP, Eisen AZ, Bauer EA, Jeffrey JJ (1978) Human skin fibroblast collagenase: Chemical properties of precursor and active forms. Biochemistry 17:2331–2337

    Article  PubMed  CAS  Google Scholar 

  • Tartakoff AM (1983) Perturbation of vesicular traffic with the carboxylic ionophore monensin. Cell 32:1026–1028

    Article  PubMed  CAS  Google Scholar 

  • Virca GD, Mallya RK, Pepys MB, Schnebli HP (1982) Quantitation of human leukocyte elastase, cathepsin G, α-2-macroglobulin and α-1-proteinase inhibitor in osteoarthritis and rheumatoid arthritis synovial fluids. Ad Exp Med Biol 167:345–357

    Google Scholar 

  • Welgus HG, Jeffrey JJ, Eisen AZ, Roswit WT, Stricklin GP (1985) Human skin fibroblast collagenase: Interaction with substrate and inhibitor. Collagen Rel Res 5:167–179

    CAS  Google Scholar 

  • Woolley DE, Crossley MJ, Evanson JM (1977) Collagenase at sites of cartilage erosion in the rheumatoid joint. Arthritis Rheum 20:1231–1239

    Article  PubMed  CAS  Google Scholar 

  • Woolley DE, Harris ED Jr, Mainardi CL, Brinckerhoff CE (1978) Collagenase immunolocalization in cultures of rheumatoid synovial cells. Science 200:773–775

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pathology, School of Medicine, Kanazawa University, 13-1 Takara-machi, 920, Kanazawa, Ishikawa, Japan

    Yasunori Okada & Isao Nakanishi

  2. Department of Orthopedic Surgery, School of Medicine, Kanazawa University, Kanazawa, Japan

    Yukio Gonoji

  3. Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas

    Hideaki Nagase

  4. Department of Biochemistry, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan

    Taro Hayakawa

Authors
  1. Yasunori Okada
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yukio Gonoji
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Isao Nakanishi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hideaki Nagase
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Taro Hayakawa
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Okada, Y., Gonoji, Y., Nakanishi, I. et al. Immunohistochemical demonstration of collagenase and tissue inhibitor of metalloproteinases (TIMP) in synovial lining cells of rheumatoid synovium. Virchows Archiv B Cell Pathol 59, 305–312 (1990). https://doi.org/10.1007/BF02899418

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02899418

Key words

Search

Navigation