Summary
Monoclonal antibodies have been used to study the presence and distribution of various components of the proteoglycan molecule in the intervertebral disc and cartilage endplate. Link protein, hyaluronic acid binding region, keratan sulphate and chondroitin 4- and 6-sulphate have been investigated in tissues from humans and other mammals. Exposure of the carbohydrate and protein epitopes was enhanced by chondroitinase and trypsin pretreatment respectively. The degree of immunoreactivity varied with location, being greater in the nucleus pulposus than the annulus fibrosus with least reactivity in the cartilage endplate. In addition, there was increased staining in the pericellular domains, particularly in adult tissues. Areas of ectopic calcification exhibited very different immunoreactivity, depending on the type of calcium salt present. Calcium hydroxyapatite deposits showed greater staining for 8A4 (link protein), while calcium pyrophosphate deposits demonstrated greater staining for 3B3(-), 7D4(-) and 3D5 than the surrounding non-calcified matrix. Staining for chondroitin sulphate isomer epitopes 3B3(-) and 7D4(-), indicative of modified chondroitin sulphate chains, was greater in human tissues of degenerate than non-degenerate appearance. This suggests that expression of these epitopes may be an indicator of disease and subsequent reparative procedures in intervertebral disc and cartilage endplate, similar to that seen in articular cartilage degeneration.
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References
Aspden, R. M., Hickey, D. S. & Hukins, D. W. L. (1981) Determination of collagen fibril orientation in the cartilage of vertebral endplate. Conn. Tissue Res. 9, 83–7.
Brandt, K. D. (1989) A pessimistic view of serologic markers for diagnosis and management of osteoarthritis. Biochemical immunologic and clinicopathologic barriers. J. Rheum. 16, 39–42.
Byers, S., Caterson, B., Hopwood, J. J. & Foster, B. K. (1992) Immunolocation analysis of glycosaminoglycans in the human growth plate. J. Histochem. Cytochem. 40, 275–82.
Carney, S. L., Billingham, M. E. J., Caterson, B., Ratcliffe, A., Bayliss, M. T., Hardingham, T. E. & Muir, H. (1992) Changes in proteoglycan turnover in experimental canine osteoarthritic cartilage. Matrix 12, 137–47.
Caterson, B. (1991) Immunological aspects of markers of joint disease. J. Rheum. 18, 19–23.
Caterson, B., Christner, J. E. & Baker, J. R. (1983) Identification of a monoclonal antibody that specifically recognizes corneal and skeletal keratan sulphate. J. Biol. Chem. 258, 8848–54.
Caterson, B., Baker, J. R., Christner, J. E., Lee, Y. & Lentz, M. (1985a) Monoclonal antibodies as probes for determining the microheterogeneity of the link proteins of cartilage proteoglycan. J. Biol. Chem. 260, 11 348–56.
Caterson, B., Christner, J. E., Baker, J. R. & Couchman, J. R. (1985b) Production and characterization of monoclonal antibodies directed against connective tissue proteoglycans. FASEB 44, 386–93.
Caterson, B., Calabro, T., Donohue, P. J. & Jahnke, M. R. (1986) In Articular Cartilage Biochemistry (edited by Kuettner, K.), pp. 59–73. New York: Raven Press.
Caterson, B., Griffin, J., Mahmoodian, F. & Sorrell, J. M. (1990a) Monoclonal antibodies against chondroitin sulphate isomers: their use as probes for investigating proteoglycan metabolism. Biochem. Soc. Trans. 18, 820–3.
Caterson, B., Mahmoodian, F., Sorrell, J. M., Hardingham, T. E., Bayliss, M. T., Carney, S. L., Ratcliffe, A. & Muir, H. (1990b) Modulation of native chondroitin sulphate structure in tissue development and in disease. J. Cell Science 97, 411–7.
Caterson, B., Hughes, C. E., Johnstone, B. & Mort, J. S. (1992) Immunological markers of cartilage proteoglycan metabolism in animal and human osteoarthritis. In Articular Cartilage and Osteoarthritis (edited by Kuettner, K. E., et al.), pp. 415–27. New York: Raven Press.
Couchman, J. R., Caterson, B., Christner, J. E. & Baker, J. R. (1984) Mapping by monoclonal antibody detection of glycosaminoglycans in connective tissue. Nature 307, 650–2.
Fosang, A. J. & Hardingham, T. E. (1991) 1-C-6 epitope in cartilage proteoglycan G2 domain is masked by keratan sulphate. Biochem. J. 273, 369–73.
Galante, J. O. (1967) Tensile properties of the human lumbar annulus fibrosus. Acta Orthop. Scand. (suppl) 100, 30–1.
Inerot, S. & Axelsson, I. (1991) Structure and composition of proteoglycans from human annulus fibrosus. Conn. Tissue Res. 26, 47–63.
Johnstone, B., Markopoulos, M., Neame, P. & Caterson, B. (1993) Identification and characterization of glycanated and non-glycanated forms of biglycan and decorin in the human intervertebral disc. Biochem. J. 292, 661–6.
Modis, L. (1990) Structure of the hyaline cartilage matrix. In Organisation of the Extracellular Matrix: a Polarization Microscopic Approach (edited by Modis, L.), pp. 75–97. Boca Raton: CRC Press.
Poole, A. R., Pidoux, I. & Rosenberg, L. (1982) Role of proteoglycans in endochondral ossification: immunofluorescent localization of link protein and proteoglycan monomer in bovine fetal epiphyseal growth plate. J. Cell. Biol. 92, 249–60.
Poole, C. A., Wotton, S. F. & Duance, V. C. (1988) Localisation of type IX collagen in chondrons isolated from porcine articular cartilage and rat chondrosarcoma. Histochem. J. 20, 567–74.
Ratcliffe, A., Fryer, P. R. & Hardingham, T. E. (1984) The distribution of aggregating proteoglycans in articular cartilage: comparison of quantitative immunoelectron microscopy with radioimmunoassay and biochemical analysis. J. Histochem. Cytochem. 32, 193–201.
Ratcliffe, A., Shurety, W. & Caterson, B. (1993) The quantitation of a native chondroitin sulphate epitope in synovial fluid lavages and articular cartilage from canine experimental osteoarthritis and disuse atrophy. Arthritis Rheum. 36, 543–51.
Roberts, S., Menage, J. & Urban, J. P. (1989) Biochemical and structural properties of the cartilage endplate and its relation to the intervertebral disc. Spine 14, 166–74.
Roberts, S., Menage, J., Duance, V., Wotton, S. & Ayad, S. (1991) Collagen types around the cells of the intervertebral disc and cartilage endplate: an immunolocalisation study. Spine 16, 1030–8.
Roberts, S., Menage, J. & Eisenstein, S. M. (1993) The cartilage endplate in scoliosis: Calcification and other sequelae. J. Orthop. Res. 11, 747–57.
Scott, J. T. (1986) Gout. In Copeman's Textbook of the Rheumatic Diseases (edited by Scott, J. T.), p. 895. Edinburgh: Churchill Livingstone.
Slater, R. R., Caterson, B., Lachiewicz, P. & Bayliss, M. T. (1992) The occurrence of 3-B-3(-) and 7-D-4 epitopes as markers of arthritis in humans. Proc. Orthop. Res. Soc. 38, 277.
Sorrell, J. M., Mahmoodian, F., Schafer, I. A., Davis, B. & Caterson, B. (1990) Identification of monoclonal antibodies that recognise novel epitopes in native chondroitin/dermatan sulphate glycosaminoglycan chains: their use in mapping functionally distinct domains of human skin. J. Histochem. Cytochem. 38, 393–402.
Urban, J. & Maroudas, A. (1980) The chemistry of the intervertebral disc in relation to its physiological function and requirements. Clin. Rheum. Dis. 6, 51–76.
Visco, D. M., Johnstone, B., Hill, M. A., Jolly, G. A. & Caterson, B. (1993) Immunohistochemical analysis of 3-B-3(-) and 7-D-4 epitope expression in canine osteoarthritis. Arthritis Rheum. 36, 1718–25.
Yada, T., Suzuki, S., Kobayashi, K., Kobayashi, M., Hoshino, T., Horie, K. & Kimata, K. (1990) Occurrence in chick embryo vitreous humor of a type IX collagen proteoglycan with an extraordinarily large chondroitin sulphate chain and short α1 polypeptide. J. Biol. Chem. 265, 6992–9.
Yamagata, T., Saito, H., Habuchi, O. & Suzuki, S. (1968) Purification and properties of bacterial chondroitinases and chondrosulfatases. J. Biol. Chem. 243, 1523–35.
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Roberts, S., Caterson, B., Evans, H. et al. Proteoglycan components of the intervertebral disc and cartilage endplate: an immunolocalization study of animal and human tissues. Histochem J 26, 402–411 (1994). https://doi.org/10.1007/BF00160052
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DOI: https://doi.org/10.1007/BF00160052