TNF receptor agonists induce distinct receptor clusters to mediate differential agonistic activity

• Published in: Communications biology Vol. 4; no. 1; p. 772
• Main Authors: Yu, Xiaojie, James, Sonya, Felce, James H., Kellermayer, Blanka, Johnston, David A., Chan, H. T. Claude, Penfold, Christine A., Kim, Jinny, Inzhelevskaya, Tatyana, Mockridge, C. Ian, Watanabe, Yasunori, Crispin, Max, French, Ruth R., Duriez, Patrick J., Douglas, Leon R., Glennie, Martin J., Cragg, Mark S.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 23.06.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 13/1; 13/109; 13/31; 14/1; 14/19; 14/63; 631/250/2503; 631/57/2267; 64/60; 82/1; 82/103; Agonists; Animals; Antibodies, Monoclonal - pharmacology; Antibody Affinity; Antitumor activity; Biology; Biomedical and Life Sciences; CD40 antigen; CD40 Antigens - agonists; CD40 Antigens - chemistry; Cell Line; Humans; Immunoglobulin G - pharmacology; Life Sciences; Mice; Microscopy, Confocal; Monoclonal antibodies; Receptor density; Receptors, OX40 - agonists; Receptors, Tumor Necrosis Factor - agonists; Receptors, Tumor Necrosis Factor - chemistry; Tumor necrosis factor; Tumor Necrosis Factor Receptor Superfamily, Member 9 - agonists; Tumor necrosis factor receptors; Tumor necrosis factor-TNF
• ISSN: 2399-3642; 2399-3642
• DOI: 10.1038/s42003-021-02309-5

Monoclonal antibodies (mAb) and natural ligands targeting costimulatory tumor necrosis factor receptors (TNFR) exhibit a wide range of agonistic activities and antitumor responses. The mechanisms underlying these differential agonistic activities remain poorly understood. Here, we employ a panel of experimental and clinically-relevant molecules targeting human CD40, 4-1BB and OX40 to examine this issue. Confocal and STORM microscopy reveal that strongly agonistic reagents induce clusters characterized by small area and high receptor density. Using antibody pairs differing only in isotype we show that hIgG2 confers significantly more receptor clustering than hIgG1 across all three receptors, explaining its greater agonistic activity, with receptor clustering shielding the receptor-agonist complex from further molecular access. Nevertheless, discrete receptor clustering patterns are observed with different hIgG2 mAb, with a unique rod-shaped assembly observed with the most agonistic mAb. These findings dispel the notion that larger receptor clusters elicit greater agonism, and instead point to receptor density and subsequent super-structure as key determinants. Yu et al examined a panel of both experimental and clinically-relevant TNF agonists in order to advance our understanding of the mechanisms underlying their varying activities and anti-tumor responses. They demonstrated that agonists with greater activity induced higher density receptor clustering and specific super-structures as opposed to simply larger receptor clusters.