title: Detection of collagenase-induced damage of collagen by 9A4, a monoclonal C-terminal neoepitope antibody.
abstract: To determine whether the collagen network is compromised by collagenase during acute inflammation, a monoclonal antibody (9A4) was developed with specificity for the C-terminal neoepitope sequence generated by collagenase-cleavage of type II collagen (Gly-Pro-Pro-Gly-Pro-Gln-Gly-COOH). 9A4 was shown to detect the collagen collagenase-cleavage neoepitope with a K = 1.7 x 10(-7) M (type II) and K = 2 x 10(-6) M (type I). It does not recognize uncleaved native or denatured collagen. Articular cartilage from control animals is unstained by 9A4. During acute inflammation elicited in hamsters by intra-articular LPS, positive staining for the 9A4 neoepitope indicated the collagen was damaged. Wheel running exercise was used to apply stress to control cartilage and cartilage from animals with damaged collagen. After 6 months of running, the cartilage from normal animals was unaffected. By contrast, in the group with damaged collagen, the cartilage was fibrillated in all animals and in half of those, the cartilage failed and bony eburnation resulted.
authors.affiliation: Department of Cancer, Immunology, and Infectious Diseases, Pfizer Central Research Division, Pfizer Inc, Groton, CT 06340, USA. otteri@pfizer.com
authors.forename: I G
authors.initials: IG
authors.lastname: Otterness
authors.forename: J T
authors.initials: JT
authors.lastname: Downs
authors.forename: C
authors.initials: C
authors.lastname: Lane
authors.forename: M L
authors.initials: ML
authors.lastname: Bliven
authors.forename: H
authors.initials: H
authors.lastname: Stukenbrok
authors.forename: D N
authors.initials: DN
authors.lastname: Scampoli
authors.forename: A J
authors.initials: AJ
authors.lastname: Milici
authors.forename: P S
authors.initials: PS
authors.lastname: Mézes
mesh_terms: D000208:Acute Disease; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000918:Antibody Specificity; D002358:Cartilage, Articular / Q000276:immunology / Q000473:pathology; D003094:Collagen / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D017364:Collagenases / Q000378:metabolism*; D006224:Cricetinae; D000939:Epitopes; D005260:Female; D007150:Immunohistochemistry; D008070:Lipopolysaccharides; D008647:Mesocricetus; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D010007:Osteochondritis / Q000139:chemically induced / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D005082:Physical Exertion; D020349:Surface Plasmon Resonance
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D008070:Lipopolysaccharides; D003094:Collagen; D017364:Collagenases