Detection of collagenase-induced damage of collagen by 9A4, a monoclonal C-terminal neoepitope antibody

Otterness, Ivan G.; Downs, James T.; Lane, Caryl; Bliven, Marcia L.; Stukenbrok, H; Scampoli, D N; Milici, Anthony J.; Mézes, Peter S.F.

doi:10.1016/s0945-053x(99)00026-8

Cited by 57 publications

(49 citation statements)

References 37 publications

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“…Several immunoassays for cartilage CII fragments have been described, which include targeting denatured triplehelical domain epitopes (51), collagenase-cleavage prod- ucts from the three-quarter-length locus (25,52), and telopeptide domain epitopes (27). A CII degradation marker is attractive because it is the most abundant protein in the tissue, it normally turns over extremely slowly, and its overt degradation is thought to mark a critical, perhaps irreversible, stage in the process of joint destruction.…”

Section: Discussionmentioning

confidence: 99%

The release of crosslinked peptides from type II collagen into human synovial fluid is increased soon after joint injury and in osteoarthritis

Lohmander¹,

Atley²,

Pietka³

et al. 2003

Arthritis & Rheumatism

234

165

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Objective. To determine concentrations of crosslinked C-telopeptide fragments of type II collagen (CTX-II) in synovial fluid (SF) from patients with joint injury, osteoarthritis (OA), or other knee arthritides.Methods. Two study groups were used: a crosssectional group, which included healthy-knee volunteers (reference group [REF]) and patients with pseudogout (PPA), an anterior cruciate ligament tear with or without a meniscus tear (INJ), or primary knee OA (POA); and a longitudinal group, which included patients with arthroscopic cartilage changes or septic arthritis. CTX-II was quantified by competition enzyme-linked immunosorbent assay (ELISA) based on a monoclonal antibody that recognized the C-terminus of the peptide EKGPDP as a proteolytic neoepitope. Aggrecan fragments, matrix metalloproteinases 1 and 3, and tissue inhibitor of metalloproteinases 1 were determined by ELISAs.Results. Concentrations of CTX-II in SF were higher in patients with PPA, INJ, and POA than in the REF group (P < 0.001). After joint injury, mean levels of CTX-II in SF were increased above REF levels at all time intervals (P < 0.001), and were highest within hours after trauma. In those in the longitudinal study group with joint cartilage damage, variation coefficients for CTX-II were 81% (between patients) and 64% (within patient), monitored over 1 year. In a patient with septic arthritis, SF CTX-II increased at the onset of symptoms, and peaked 30-fold higher than the baseline. Concentrations of all biomarkers decreased with successful treatment.Conclusion. This is the first report to describe the release into SF of soluble molecular fragments specific for the degradation of mature, crosslinked, type II collagen (CII) in human OA and joint injury. The results provide strong evidence that the integrity of the CII network of cartilage is compromised soon after joint injury and in arthritis. This early degradation of CII may represent an important treatment target.Osteoarthritis (OA) and joint injury are characterized by remodeling and degradation of cartilage, bone, and other joint tissues. The normally very slow turnover rate of cartilage matrix (1) is increased, as observed by several techniques in both animal OA models and human OA. Phasic changes in both synthesis and degradation of collagen and other matrix molecules (2-6) are associated with altered tissue structure and material properties (7-9).Increased joint tissue turnover after injury and in OA can be detected by a release of molecules and molecular fragments into synovial fluid (SF), blood, and urine. For example, the stimulated synthesis of type II collagen (CII) results in higher levels of CII C-propeptide in SF (10,11). Stimulated aggrecan synthesis results in more aggrecan fragments carrying the 846 epitope (12). Higher synovial concentrations of matrix metalloproteinases (MMPs), such as MMP-1 and MMP-3, and proteinase activity after joint injury and in OA are consistent with the observed expression of these

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Section: Discussionmentioning

confidence: 99%

The release of crosslinked peptides from type II collagen into human synovial fluid is increased soon after joint injury and in osteoarthritis

Lohmander¹,

Atley²,

Pietka³

et al. 2003

Arthritis & Rheumatism

234

165

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“…Upon cleavage of collagen by a collagenase, this epitope is exposed and staining can be observed by immunohistochemistry (29). Staining of this 9A4 neoepitope lagged behind loss of proteoglycan staining and was first observable by ∼21 d in the elbow (Fig.…”

Section: Molecular Characterization Of Cartilage Matrix Degradation Inmentioning

confidence: 99%

NFATc1 and NFATc2 repress spontaneous osteoarthritis

Greenblatt

Ritter²,

Wright

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Osteoarthritis (OA) was once viewed originally as a mechanical disease of "wear and tear," but advances made during the past two decades suggest that abnormal biomechanics contribute to active dysregulation of chondrocyte biology, leading to catabolism of the cartilage matrix. A number of signaling and transcriptional mechanisms have been studied in relation to the regulation of this catabolic program, but how they specifically regulate the initiation or progression of the disease is poorly understood. Here, we demonstrate that cartilage-specific ablation of Nuclear factor of activated T cells c1 (Nfatc1) in Nfatc2 −/− mice leads to early onset, aggressive OA affecting multiple joints. This model recapitulates features of human OA, including loss of proteoglycans, collagen and aggrecan degradation, osteophyte formation, changes to subchondral bone architecture, and eventual progression to cartilage effacement and joint instability. Consistent with the notion that NFATC1 is an OA-suppressor gene, NFATC1 expression was significantly down-regulated in paired lesional vs. macroscopically normal cartilage samples from OA patients. The highly penetrant, early onset, and severe nature of this model make it an attractive platform for the preclinical development of treatments to alter the course of OA. Furthermore, these findings indicate that NFATs are key suppressors of OA, and regulating NFATs or their transcriptional targets in chondrocytes may lead to novel disease-modifying OA therapies.

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“…For 9A4 immunohistochemistry (39), sections from the stifle joint were stained using DAKO ARK kit (DAKO, Carpinteria, CA) according to the manufacturer's directions. Ag retrieval was performed with Decal Solution (Biogenex, San Ramon, CA) at room temperature for 30 min.…”

Section: Mab-induced Arthritismentioning

confidence: 99%

“…A modified Mankin score (40) was calculated based on cartilage structure, cellularity, safranin-O staining of mucopolysaccharides, and synovial inflammation/hyperplasia. In addition, to assess collagen degradation, sections were stained with a mAb (9A4) that specifically recognizes the neoepitope generated by collagenase digestion (39). Lesions observed in the articular cartilage of the stifle and hock joints recovered from wild-type animals included slight to moderate reductions in glycosaminoglycans (extending to the tidemark of the articular cartilage), which correlated inversely with increased 9A4 immunohistochemical staining (Fig.…”

Section: Mab-induced Arthritismentioning

confidence: 99%

“…This latter model is dependent on IL-1, and inhibitors of the production and/or activity of this cytokine suppress disease outcomes (55,56). At the histological level, mAb-induced arthritis led to a disruption of the normal cartilage architecture, loss of proteoglycan content, synovial inflammation, and the appearance of collagen cleavage products as detected with the mAb 9A4 (39). The time required to achieve disease in the monoclonal arthritis model is reduced relative to that required in the collagen-induced model, but the severity of the disease appears comparable in the two formats.…”

Section: Figurementioning

confidence: 99%

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Absence of the P2X7 Receptor Alters Leukocyte Function and Attenuates an Inflammatory Response

Labasi

Petrushova

Donovan

et al. 2002

The Journal of Immunology

450

378

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When challenged with extracellular ATP, leukocytes respond and activate processes attributed to the P2X7 receptor (P2X7R), an unusual ligand-gated ion channel. To prove P2X7R involvement, blood samples from P2X7R-deficient mice were characterized. Monocytes and lymphocytes associated with wild-type blood responded to ATP and underwent volume/shape changes and shed L-selectin. In contrast, leukocytes from P2X7R-deficient animals demonstrated no change in physical properties or L-selectin expression following ATP challenge. Blood stimulated with LPS or ATP individually generated minimal quantities of the leaderless polypeptide IL-1β, but sequential treatment of wild-type, but not P2X7R-deficient, blood with LPS and ATP yielded large amounts of cell-free cytokine. Based on these differences, wild-type and P2X7R-deficient animals were compared following induction of monoclonal anti-collagen-induced arthritis. Ab-treated wild-type animals subsequently challenged with LPS developed inflamed, swollen paws; their joint cartilage demonstrated lesions, loss of proteoglycan content, and the presence of collagen degradation products. P2X7R-deficient animals subjected to the same challenge were markedly less affected; both the incidence and severity of disease were reduced. These data indicate that ATP does act via the P2X7R to affect leukocyte function and that the P2X7R can serve as an important component of an in vivo inflammatory response.

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Detection of collagenase-induced damage of collagen by 9A4, a monoclonal C-terminal neoepitope antibody

Cited by 57 publications

References 37 publications

The release of crosslinked peptides from type II collagen into human synovial fluid is increased soon after joint injury and in osteoarthritis

The release of crosslinked peptides from type II collagen into human synovial fluid is increased soon after joint injury and in osteoarthritis

NFATc1 and NFATc2 repress spontaneous osteoarthritis

Absence of the P2X7 Receptor Alters Leukocyte Function and Attenuates an Inflammatory Response

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