Comparative studies of collagenase and stromelysin-1 expression by rheumatoid synoviocytes in vitro.
Matrix metalloproteinases such as collagenase and stromelysin are recognised as important cartilage-degrading enzymes in the pathophysiology of rheumatoid arthritis. Synovial fibroblasts and macrophages are the major cellular components of rheumatoid synovium, but the regulation and relative expression of collagenase and stromelysin by these two cell types remains uncertain. Using in vitro cultures of adherent rheumatoid synovial cells we have examined the coordinate or separate expression of collagenase and stromelysin-1 by dual immunolocalisation and Western blotting techniques. Synovial fibroblasts, when activated by macrophage-derived products in primary culture or by interleukin-1/phorbol myristate acetate in subcultures, released significant quantities of collagenase and stromelysin in their inactive, precursor forms. The ratio of released procollagenase: prostromelysin varied between different synovial cell preparations. Dual immunolocalisation studies demonstrated both coordinate and separate expression of the two enzymes by single cells. Approximately 80% of the activated fibroblasts, especially those with stellate morphology, showed co-expression of both enzymes. By contrast synovial macrophages had a modest or negligible capacity to elaborate either enzyme under the same in vitro conditions. In many fibroblastic cells both collagenase and stromelysin were co-localised to the perinuclear Golgi region and the same cytoplasmic compartments. Vesicular structures appear to provide intracellular transport for both enzymes to sites of secretion. Both enzymes showed preferential pericellular binding to a collagenous substratum rather than any association with the plasma membrane/cell surface.
Tetlow
L C
LC
Department of Medicine, University Hospital of South Manchester, West Didsbury, UK.
Lees
M
M
Woolley
D E
DE
D001172:Arthritis, Rheumatoid / Q000201:enzymology*; D015153:Blotting, Western; D002478:Cells, Cultured; D017364:Collagenases / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000201:enzymology*; D006801:Humans; D007124:Immunoenzyme Techniques; D020781:Matrix Metalloproteinase 1; D019278:Matrix Metalloproteinase 3; D008666:Metalloendopeptidases / Q000032:analysis*; D013583:Synovial Membrane / Q000166:cytology / Q000201:enzymology*
D017364:Collagenases; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3; D020781:Matrix Metalloproteinase 1