Inhibition of transmembrane TNF-α shedding by a specific antibody protects against septic shock

• Published in: Cell death & disease Vol. 10; no. 8; pp. 586 - 17
• Main Authors: Li, Chenxi, Gu, Haiyan, Yu, Mingxia, Yang, Peng, Zhang, Meng, Ba, Hongping, Yin, Yue, Wang, Jing, Yin, Bingjiao, Zhou, Xiaoxi, Li, Zhuoya
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 05.08.2019; Springer Nature B.V
• Subjects: 13/1; 13/109; 13/21; 13/31; 13/95; 631/154/556; 631/250/127/1220; 631/250/2504/342; 631/250/256/1980; 631/250/262/2106/2108; 64/110; 64/60; 82/29; 82/80; Animals; Antibodies; Antibodies, Monoclonal - pharmacology; Antibodies, Monoclonal - therapeutic use; Biochemistry; Biodegradation; Biomedical and Life Sciences; Cecum; Cell Biology; Cell Culture; Cell Membrane - metabolism; Disease Models, Animal; Gene Knockout Techniques; HEK293 Cells; Humans; Immunology; Inflammation; Interferon; Interferon regulatory factor; Interferon regulatory factor 3; Interleukin 6; Internalization; Life Sciences; Lipopolysaccharides; Lipopolysaccharides - pharmacology; Macrophages; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Monoclonal antibodies; Monocytes; NIH 3T3 Cells; Nitric oxide; Protective Agents - pharmacology; Protective Agents - therapeutic use; RAW 264.7 Cells; Receptors, Tumor Necrosis Factor, Type I - genetics; Receptors, Tumor Necrosis Factor, Type II - genetics; Sepsis; Septic shock; Shock, Septic - chemically induced; Shock, Septic - drug therapy; THP-1 Cells; TLR4 protein; Toll-Like Receptor 4 - metabolism; Toll-like receptors; Transfection; Tumor Necrosis Factor-alpha - antagonists & inhibitors; Tumor Necrosis Factor-alpha - genetics; Tumor Necrosis Factor-alpha - metabolism; Tumor necrosis factor-α; β-Interferon
• ISSN: 2041-4889; 2041-4889
• DOI: 10.1038/s41419-019-1808-6

Transmembrane TNF-α (tmTNF-α) and secretory TNF-α (sTNF-α) display opposite effects in septic shock. Reducing tmTNF-α shedding can offset the detrimental effects of sTNF-α and increase the beneficial effect of tmTNF-α. We previously developed a monoclonal antibody that is specific for tmTNF-α and does not cross-react with sTNF-α. In this study, we show that this antibody can specifically suppress tmTNF-α shedding by competing with a TNF-α converting enzyme that cleaves the tmTNF-α ectodomain to release sTNF-α. This tmTNF-α antibody significantly inhibited LPS-induced secretion of interleukin (IL)-1β, IL-6, interferon-β, and nitric oxide by monocytes/macrophages, and protected mice from septic shock induced by lipopolysaccharide (LPS) or cecal ligation and puncture, while reducing the bacterial load. The mechanism associated with the protective effect of this tmTNF-α antibody involved promotion of LPS-induced toll-like receptor 4 (TLR4) internalization and degradation by recruiting Triad3A to TLR4. Moreover, the tmTNF-α antibody inhibited LPS-induced activation of nuclear factor-κB and interferon regulatory factor 3 pathways by upregulating expression of A20 and monocyte chemotactic protein-induced protein 1. Similarly, treatment of macrophages with exogenous tmTNF-α suppressed LPS/TLR4 signaling and release of proinflammatory cytokines, indicating that increased levels of tmTNF-α promoted by the antibody contributed to its inhibitory effect. Thus, use of this tmTNF-α antibody for specific suppression of tmTNF-α shedding may be a promising strategy to treat septic shock.