Efficient induction of arthritis in mice by an arthritogenic 4-clone cocktail of anti-type II collagen monoclonal antibodies recognizing different epitopes of the antigen.

We previously reported that a combination of 4 monoclonal antibodies (mAbs) (cocktail A) to type II collagen (CII), including immunoglobulin G (IgG)2b (C2B-9 and C2B-14) and IgG2a (C2A-7 and C2A-12), induced arthritis in DBA/1J mice. In this study, we found that C2B-9 and C2A-7 as well as C2B-14 and C2A-12 recognized the same or similar epitopes on CII. Based on these data, we hypothesized that the combination of more than 3 mAbs recognizing different epitopes on CII may more efficiently induce arthritis. Therefore, in addition to C2B-9 and C2B-14, which show high binding activity to CII compared with C2A-7 and C2A-12, we developed two more mAbs including IgG2b (C2B-17) and IgG2a (C2A-16), to make a new cocktail (cocktail B) consisting of these 4 mAbs. To compare the ability of cocktail B to induce arthritis with cocktail A, DBA/1J mice were injected with these cocktails. The results showed that cocktail B was able to induce more severe arthritis than cocktail A, especially more markedly affecting rear paws. Histologically, there was more marked proliferation of synovial tissues, massive infiltration by inflammatory cells, and severe destruction of cartilage and bone in mice treated with cocktail B than with cocktail A. Collectively, the new combination of 4 mAbs recognizing different respective epitopes appears to effectively induce arthritis in mice. Thus, the results may provide insights into the selection of mAbs associated with the development of arthritis.

Arthrogenicity of type II collagen monoclonal antibodies associated with complement activation and antigen affinity.

BACKGROUND: The collagen antibody-induced arthritis (CAIA) model, which employs a cocktail of monoclonal antibodies (mAbs) to type II collagen (CII), has been widely used for studying the pathogenesis of autoimmune arthritis. In this model, not all mAbs to CII are capable of inducing arthritis because one of the initial events is the formation of collagen-antibody immune complexes on the cartilage surface or in the synovium, and subsequent activation of the complement by the complexes induces arthritis, suggesting that a combination of mAbs showing strong ability to bind mouse CII and activate the complement may effectively induce arthritis in mice. In the present study, we examined the relationship between the induction of arthritis by the combination of IgG2a (CII-6 and C2A-12), IgG2b (CII-3, C2B-14 and C2B-16) and IgM (CM-5) subclones of monoclonal antibodies (mAb) of anti-bovine or chicken CII and the ability of mAbs to activate complement and bind mouse CII. METHODS: DBA/1J mice were injected with several combinations of mAbs followed by lipopolysaccharide. Furthermore, the ability of mAbs to activate the complement and bind mouse CII was examined by ELISA. RESULTS: First, DBA/1J mice were injected with the combined 4 mAbs (CII-3, CII-6, C2B-14, and CM-5) followed by lipopolysaccharide, resulting in moderate arthritis. Excluding one of the mAbs, i.e., using only CII-3, CII-6, and C2B-14, induced greater inflammation of the joints. Next, adding C2A-12 but not C2B-16 to these 3 mAbs produced more severe arthritis. A combination of five clones, consisting of all 5 mAbs, was less effective. Histologically, mice given the newly developed 4-clone cocktail had marked proliferation of synovial tissues, massive infiltration by inflammatory cells, and severe destruction of cartilage and bone. Furthermore, 4 of the 6 clones (CII-3, CII-6, C2B-14, and C2A-12) showed not only a strong cross-reaction with mouse CII but also marked activation of the complement in vitro. CONCLUSION: The combination of 4 mAbs showing strong abilities to activate the complement and bind mouse CII effectively induced arthritis in DBA/1J mice. This in vitro system may be useful for the selection of mAbs associated with the development of arthritis.