Bivalent structure of a TNFR2-selective and agonistic TNF-α mutein Fc-fusion protein enhances the expansion activity of regulatory T cells

• Published in: Scientific reports 2023-08, Vol.13 (1), p.13762-13762, Article 13762
• Main Authors: Inoue, Masaki, Tsuji, Yuta, Ueno, Reira, Miyamoto, Daisuke, Tanaka, Keisuke, Moriyasu, Yuka, Shibata, Saya, Okuda, Mei, Ando, Daisuke, Abe, Yasuhiro, Kamada, Haruhiko, Tsunoda, Shin-ichi
• Format: Article
• Language: English
• Subjects: 631/250/251; 631/250/256; 631/61/51; Agonistic behavior; Allergies; Animal models; Autoimmune diseases; CD25 antigen; CD4 antigen; Cell activation; Cell proliferation; Contact dermatitis; Fc receptors; Foxp3 protein; Fusion protein; Humanities and Social Sciences; Hypersensitivity; Immunoglobulin G; Immunological tolerance; Immunoregulation; Immunosuppressive agents; Leukocytes (mononuclear); Lymphocytes; Lymphocytes T; multidisciplinary; Peripheral blood mononuclear cells; Science; Science (multidisciplinary); Tumor necrosis factor receptors; Tumor necrosis factor-TNF; Tumor necrosis factor-α
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-023-40925-9

Recently, TNF receptor type 2 (TNFR2) signaling was found to be involved in the proliferation and activation of regulatory T cells (Tregs), a subpopulation of lymphocytes that suppress immune responses. Tregs mediate peripheral immune tolerance, and the disruption of their functions causes autoimmune diseases or allergy. Therefore, cell expanders or regulators of Tregs that control immunosuppressive activity can be used to treat these diseases. We focused on TNFR2, which is preferentially expressed on Tregs, and created tumor necrosis factor-α (TNF-α) muteins that selectively activate TNFR2 signaling in mice and humans, termed R2agoTNF and R2-7, respectively. In this study, we attempted to optimize the structure of muteins to enhance their TNFR2 agonistic activity and stability in vivo by IgG-Fc fusion following single-chain homo-trimerization. The fusion protein, scR2agoTNF-Fc, enhanced the expansion of CD4 + CD25 + Tregs and CD4 + Foxp3 + Tregs and contributed to their immunosuppressive activity ex vivo and in vivo in mice. The prophylactic administration of scR2agoTNF-Fc suppressed inflammation in contact hypersensitivity and arthritis mouse models. Furthermore, scR2-7-Fc preferentially expanded Tregs in human peripheral blood mononuclear cells via TNFR2. These TNFR2 agonist-Fc fusion proteins, which have bivalent structures, are novel Treg expanders.