Generation and characterization of novel co-stimulatory anti-mouse TNFR2 antibodies

• Published in: Journal of immunological methods 2021-12, Vol.499, p.113173-113173, Article 113173
• Main Authors: Segués, Aina, van Duijnhoven, Sander M.J., Parade, Marc, Driessen, Lilian, Vukovic, Nataša, Zaiss, Dietmar, Sijts, Alice J.A.M., Berraondo, Pedro, van Elsas, Andrea
• Format: Article
• Language: English
• Subjects: Antibody; Costimulation; Cysteine-rich domain; Epitope; TNFR2; Treg
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/j.jim.2021.113173

Tumor necrosis factor receptor 2 (TNFR2) has gained much research interest in recent years because of its potential pivotal role in autoimmune disease and cancer. However, its function in regulating different immune cells is not well understood. There is a need for well-characterized reagents to selectively modulate TNFR2 function, thereby enabling definition of TNFR2-dependent biology in human and mouse surrogate models. Here, we describe the generation, production, purification, and characterization of a panel of novel antibodies targeting mouse TNFR2. The antibodies display functional differences in binding affinity and potency to block TNFα. Furthermore, epitope binding showed that the anti-mTNFR2 antibodies target different domains on the TNFR2 protein, associated with varying capacity to enhance CD8+ T-cell activation and costimulation. Moreover, the anti-TNFR2 antibodies demonstrate binding to isolated splenic mouse Tregs ex vivo and activated CD8+ cells, reinforcing their potential use to establish TNFR2-dependent immune modulation in translational models of autoimmunity and cancer. •We have generated a diverse library of anti-mouse TNFR2 antibodies.•Developed anti-mouse TNFR2 antibodies show binding to regulatory T cells (Tregs) and activated CD8+ cells.•Some anti-mouse TNFR2 antibodies costimulate CD8+ cells.