Immunological analysis of proteoglycan structural changes in the early stage of experimental osteoarthritic canine cartilage lesions.
Specific modifications of the proteoglycan (PG) structure of osteoarthritic (OA) dog cartilage in relation to endogenous metalloprotease activity were examined using murine anti-proteoglycan monoclonal antibodies (MoAbs). OA lesions were induced over a period of 8 weeks in crossbred dogs (Pond-Nuki model). The articular cartilage was removed and homogenized in a Tris buffer, pH 7.5, and then divided into four groups: direct PG extraction, no addition, presence of 1 mM p-aminophenyl mercuric acetate (APMA), and presence of 1 mM APMA and 10 mM o-phenanthroline, incubated for 42 h at 37 degrees C followed by PG extraction. MoAbs reactive with PG protein and carbohydrate epitopes included 1C6, 3B3, 5D4, D1B2, and m4D6. The results showed marked alterations induced by APMA activation of the endogenous metalloproteases. PG changes were apparent at at least three sites: one was either in the hyaluronic acid-binding region or between the hyaluronic-binding region and the G2 globular domain, another was between the keratan-sulfate-rich domain and the chondroitin sulfate-attachment domain, and a third was in the chondroitin sulfate-attachment domain. Constitutive metalloprotease activity resulted in less marked PG alterations with preservation of functional PG aggregation to hyaluronan.
Pelletier
J P
JP
University of Montreal, Rheumatic Disease Unit, Notre-Dame Hospital Research Center, Canada.
Martel-Pelletier
J
J
Mehraban
F
F
Malemud
C J
CJ
D000818:Animals; D000911:Antibodies, Monoclonal; D002358:Cartilage, Articular / Q000737:chemistry / Q000473:pathology* / Q000648:ultrastructure; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D004789:Enzyme Activation / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis / Q000276:immunology; D006025:Glycosaminoglycans / Q000032:analysis / Q000276:immunology; D006898:Hydroxylamines / Q000494:pharmacology; D007150:Immunohistochemistry; D008666:Metalloendopeptidases / Q000032:analysis / Q000378:metabolism; D008854:Microscopy, Electron; D010003:Osteoarthritis / Q000276:immunology* / Q000473:pathology; D010662:Phenylmercuric Acetate / Q000031:analogs & derivatives / Q000494:pharmacology; D011509:Proteoglycans / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D011863:Radioimmunoassay; D013997:Time Factors
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006025:Glycosaminoglycans; D006898:Hydroxylamines; D011509:Proteoglycans; C010024:4-aminophenylmercuriacetate; D008666:Metalloendopeptidases; D010662:Phenylmercuric Acetate