Cartilage Antibody Cocktail (Cab3)

Immunohistochemical staining to investigate antibody binding to cartilage in vivo

Cab3 and Cab4 cocktails are pools of three and four arthritogenic antibodies, respectively, with antibodies known to bind cartilage and initiate arthritis in several mouse strains, including the commonly used C57BL/6 strain. Biotinylated versions of the antibodies may be administered in vivo, and avidin-conjugated reagents can then be used to visualize and quantify the in vivo -bound antibodies in the tissue processed ex vivo for confocal microscopy.   

The methodology to detect the cartilage-bound antibodies may be adapted to investigate the timing and fate of the arthritogenic antibodies and may be supplemented with immunohistochemical method to detect complement deposited on the cartilage. This information may be utilized to investigate the mechanisms of arthritis development in preclinical models.

References
1. Kelkka T, Hultqvist M, Nandakumar KS, Holmdahl R. Enhancement of antibody-induced arthritis via Toll-like receptor 2 stimulation is regulated by granulocyte reactive oxygen species. Am J Pathol. 2012;181(1):141–50.
2. Hagert C, Sareila O, Kelkka T, Nandakumar KS, Collin M, Xu B, Guérard S, Bäcklund J, Jalkanen S, Holmdahl R. Chronic Active Arthritis Driven by Macrophages Without Involvement of T Cells: A Novel Experimental Model of Rheumatoid Arthritis. Arthritis Rheumatol. 2018 Aug;70(8):1343-1353.
3. Li Y, Tong D, Liang P, Lönnblom E, Viljanen J, Xu B, Nandakumar KS and Holmdahl R. Cartilage-binding antibodies initiate joint inflammation and promote chronic erosive arthritis. Arthritis Research & Therapy (2020) 22:120. (Link to the Creative Commons Licence: http://creativecommons.org/licenses/by/4.0/)
4. Kagari T, Tanaka D, Doi H, Shimozato T. Essential role of Fc gamma receptors in anti-type II collagen antibody-induced arthritis. J Immunol. 2003;170(8):4318–24.
5. Holers VM, Banda NK. Complement in the Initiation and Evolution of Rheumatoid Arthritis. Front Immunol. 2018; 9: 1057.
6. Ge C, Tong D, Liang B, Lönnblom E, Schneider N, Hagert C, Viljanen J, Ayoglu B, Stawikowska R, Nilsson P, Fields GB, Skogh T, Kastbom A, Kihlberg J, Burkhardt H, Dobritzsch D, Holmdahl R. Anti-citrullinated protein antibodies cause arthritis by cross-reactivity to joint cartilage. JCI Insight. 2017 Jul 6; 2(13): e93688.