Use of an antineoepitope antibody for identification of type-II collagen degradation in equine articular cartilage.
Abstract: To develop an antibody that specifically recognizes collagenase-cleaved type-II collagen in equine articular cartilage. Methods: Cartilage specimens from horses euthanatized for problems unrelated to the musculoskeletal system. Methods: A peptide was synthesized representing the carboxy- (C-) terminus (neoepitope) of the equine type-II collagen fragment created by mammalian collagenases. This peptide was used to produce a polyclonal antibody, characterized by western analysis for reactivity to native and collagenase-cleaved equine collagens. The antibody was evaluated as an antineoepitope antibody by ELISA, using peptides +/- an amino acid at the C-terminus of the immunizing peptide. Collagen cleavage was assayed from equine articular cartilage cultured with interleukin-1 (IL-1), +/- a synthetic MMP inhibitor, BAY 12-9566. Cartilage specimens from osteoarthritic and nonarthritic joints were compared for antibody staining. Results: An antibody, 234CEQ, recognized only collagenase-generated 3/4-length fragments of equine type-II collagen. This was a true antineoepitope antibody, as altering the C-terminus of the immunizing peptide significantly decreased competition for binding in an inhibition ELISA. The IL-1-induced release of type-II collagen fragments from articular cartilage was prevented with the MMP inhibitor. Cartilage from an osteoarthritic joint of a horse had increased staining with the 234CEQ antibody, compared with normal articular cartilage. Conclusions: We generated an antineoepitope antibody recognizing collagenase-cleaved type-II collagen of horses. This antibody detects increases in type-II collagen cleavage in diseased equine articular cartilage. The 234CEQ antibody has the potential to aid in the early diagnosis of arthritis and to monitor treatment responses.
Publication Date: 2001-07-17 PubMed ID: 11453476DOI: 10.2460/ajvr.2001.62.1031Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Antibodies
- Articular Cartilage
- Collagen
- Diagnosis
- Disease Diagnosis
- Disease Treatment
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Equine Diseases
- Equine Health
- Immunization
- Immunology
- In Vitro Research
- Inflammation
- Laboratory Methods
- Musculoskeletal System
- Osteoarthritis
- Peptides
- Veterinary Care
- Veterinary Medicine
- Veterinary Research
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research article is focused on developing an antibody that can identify the degradation of type-II collagen in equine articular cartilage, aiding in early detection and monitoring treatment of arthritis in horses.
Development of the Antibody
- The researchers synthesized a peptide which represents the carboxy- (C-) terminus (neoepitope) of the equine type-II collagen fragment generated by mammalian collagenases.
- This peptide was then used to produce a polycline antibody: once produced, the antibody was characterized via a western analysis to determine its reactivity to both native and collagenese-cleaved equine collagens.
Antineoepitope Antibody Evaluation
- The antibody was then evaluated as an antineoepitope antibody using ELISA, using peptides with and without an amino acid at the C-terminus of the immunizing peptide.
- The researchers also investigated whether collagen cleavage could be assayed from equine articular cartilage which had been exposed to interleukin-1 (IL-1), with and without a synthetic MMP inhibitor, BAY 12-9566.
Experimental Results
- The results of the experiment showed that the antibody, named 234CEQ, recognized only collagenase-generated 3/4-length fragments of equine type-II collagen, therefore showing characteristics of a true antineoepitope antibody.
- The effectiveness of the antineoepitope antibody was confirmed in an inhibition ELISA, where disruptions at the C-terminus of the immunizing peptide caused a significant decrease in competition for binding.
- The research also showed that the induced release of type-II collagen fragments from articular cartilage by IL-1 was prevented by the MMP inhibitor.
Application and Conclusion
- Articular cartilage from an osteoarthritic and non-arthritic joint was compared for antibody staining. Cartilage from osteoarthritic joint showed increased staining with the 234CEQ antibody, as opposed to normal articular cartilage.
- The results demonstrate the successful development of an antineoepitope antibody that recognizes collagenase-cleaved type-II collagen in horses.
- The newly developed 234CEQ antibody has the potential to help with early diagnosis of arthritis as well as monitor responses to treatment in equine subjects.
Cite This Article
APA
Billinghurst RC, Buxton EM, Edwards MG, McGraw MS, McIlwraith CW.
(2001).
Use of an antineoepitope antibody for identification of type-II collagen degradation in equine articular cartilage.
Am J Vet Res, 62(7), 1031-1039.
https://doi.org/10.2460/ajvr.2001.62.1031 Publication
Researcher Affiliations
- Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins 80523, USA.
MeSH Terms
- Animals
- Antibodies / immunology
- Antineoplastic Agents / immunology
- Biphenyl Compounds
- Blotting, Western / veterinary
- Cartilage, Articular / immunology
- Cartilage, Articular / metabolism
- Cartilage, Articular / pathology
- Collagen Type II / immunology
- Collagen Type II / metabolism
- Collagenases / metabolism
- Enzyme-Linked Immunosorbent Assay / veterinary
- Epitopes / immunology
- Female
- Horse Diseases / immunology
- Horse Diseases / metabolism
- Horses / immunology
- Horses / metabolism
- Immunohistochemistry / veterinary
- Matrix Metalloproteinase 3 / chemistry
- Organic Chemicals
- Osteoarthritis / immunology
- Osteoarthritis / metabolism
- Osteoarthritis / veterinary
- Peptide Fragments / immunology
- Peptide Fragments / metabolism
- Phenylbutyrates
- Proteoglycans / immunology
- Proteoglycans / metabolism
- Rabbits
Citations
This article has been cited 5 times.- Martinez RE, Leatherwood JL, Arnold CE, Glass KG, Walter KW, Valigura HC, Norton SA, White-Springer SH. Responses to an intra-articular lipopolysaccharide challenge following dietary supplementation of Saccharomyces cerevisiae fermentation product in young horses. J Anim Sci 2021 Oct 1;99(10).
- Xiao L, Majumdar R, Dai J, Li Y, Xie L, Shen FH, Jin L, Li X. Molecular Detection and Assessment of Intervertebral Disc Degeneration via a Collagen Hybridizing Peptide. ACS Biomater Sci Eng 2019 Apr 8;5(4):1661-1667.
- Birmingham JD, Vilim V, Kraus VB. Collagen biomarkers for arthritis applications. Biomark Insights 2007 Feb 7;1:61-76.
- Dhiman MS, Bader TJ, Ponjevic D, Salo PT, Hart DA, Swamy G, Matyas JR, Duncan NA. Collagen integrity of the annulus fibrosus in degenerative disc disease individuals quantified with collagen hybridizing peptide. JOR Spine 2024 Sep;7(3):e1359.
- Torga T, Suutre S, Kisand K, Aunapuu M, Arend A. Cartilage Collagen Neoepitope C2C Expression in the Articular Cartilage and Its Relation to Joint Tissue Damage in Patients with Knee Osteoarthritis. Biomedicines 2024 May 11;12(5).
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