Measurement of antibodies to collagen II by inhibition of collagen fibril formation in vitro

• Published in: Journal of immunological methods 2004-02, Vol.285 (1), p.55-61
• Main Authors: Gray, Robyn E, Seng, Natalie, Mackay, Ian R, Rowley, Merrill J
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - analysis; Arthritis, Experimental - immunology; Arthritis, Rheumatoid - immunology; Biological and medical sciences; Biopolymers - chemistry; Biopolymers - immunology; Cattle; Collagen II; Collagen Type II - chemistry; Collagen Type II - immunology; Collagen Type II - ultrastructure; Fibrillogenesis; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Humans; Hydrogen-Ion Concentration; Immunoassay - methods; Macromolecular Substances; Mice; Microscopy, Electron; Molecular immunology; Monoclonal antibodies; Self-assembly; Spectrophotometry - methods; Techniques
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/j.jim.2003.11.010

Antibodies to type II collagen (collagen II) are pathogenic in experimental collagen-induced arthritis (CIA) and possibly also in rheumatoid arthritis (RA). Hitherto, results of assays for anti-collagen II have proven to be inconsistent. We tested whether mouse monoclonal antibodies (mAbs) to collagen II inhibit the natural self-assembly of soluble triple-stranded collagen II monomers to form insoluble polymeric fibrils. A spectrophotometric assay of self-assembly was based on change in absorbance at 313 nm, observed over 0–60 min after neutralisation and warming of a solution of monomeric collagen II. Two mAbs to collagen II (CII-CI and M2.139) strongly inhibited self-assembly of collagen II but not collagen I, whereas another antibody, CII-F4, and an irrelevant control mAb did not. Notably, CII-CI and M2.139, but not CII-F4, induce arthritis on passive transfer to naı̈ve mice. The arthritogenic effects of mAbs CII-CI and M2.139 in vivo, and inhibition of collagen II self-assembly in vitro, may be attributable to interference with critical epitopes at sites essential for the stabilisation of the mature polymeric collagen II fibril, and, hence, the integrity of the entire cartilage matrix. This assay for inhibition of self-assembly of collagen II could be developed for routine measurement of anti-collagen II in body fluids as a marker of early RA, and perhaps also to distinguish populations of antibodies to collagen II that either have or lack the capacity to perpetuate arthritis.