T cell–derived tumor necrosis factor induces cytotoxicity by activating RIPK1-dependent target cell death
Nicholas Chun, Rosalind L. Ang, Mark Chan, Robert L. Fairchild, William M. Baldwin III, Julian K. Horwitz, Jesse D. Gelles, Jerry Edward Chipuk, Michelle A. Kelliher, Vasile I. Pavlov, Yansui Li, Dirk Homann, Peter S. Heeger, Adrian T. Ting
Nicholas Chun, Rosalind L. Ang, Mark Chan, Robert L. Fairchild, William M. Baldwin III, Julian K. Horwitz, Jesse D. Gelles, Jerry Edward Chipuk, Michelle A. Kelliher, Vasile I. Pavlov, Yansui Li, Dirk Homann, Peter S. Heeger, Adrian T. Ting
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Research Article Cell biology Immunology

T cell–derived tumor necrosis factor induces cytotoxicity by activating RIPK1-dependent target cell death

Abstract

TNF ligation of TNF receptor 1 (TNFR1) promotes either inflammation and cell survival by (a) inhibiting RIPK1’s death-signaling function and activating NF-κB or (b) causing RIPK1 to associate with the death-inducing signaling complex to initiate apoptosis or necroptosis. The cellular source of TNF that results in RIPK1-dependent cell death remains unclear. To address this, we employed in vitro systems and murine models of T cell–dependent transplant or tumor rejection in which target cell susceptibility to RIPK1-dependent cell death could be genetically altered. We show that TNF released by T cells is necessary and sufficient to activate RIPK1-dependent cell death in target cells and thereby mediate target cell cytolysis independently of T cell frequency. Activation of the RIPK1-dependent cell death program in target cells by T cell–derived TNF accelerates murine cardiac allograft rejection and synergizes with anti-PD1 administration to destroy checkpoint blockade–resistant murine melanoma. Together, the findings uncover a distinct immunological role for TNF released by cytotoxic effector T cells following cognate interactions with their antigenic targets. Manipulating T cell TNF and/or target cell susceptibility to RIPK1-dependent cell death can be exploited to either mitigate or augment T cell–dependent destruction of allografts and malignancies to improve outcomes.

Authors

Nicholas Chun, Rosalind L. Ang, Mark Chan, Robert L. Fairchild, William M. Baldwin III, Julian K. Horwitz, Jesse D. Gelles, Jerry Edward Chipuk, Michelle A. Kelliher, Vasile I. Pavlov, Yansui Li, Dirk Homann, Peter S. Heeger, Adrian T. Ting

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Figure 6

T cell–derived TNF drives RIPK1-dependent tumor killing and synergizes with checkpoint inhibition in response to murine melanoma.

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T cell–derived TNF drives RIPK1-dependent tumor killing and synergizes w...
(A) Quantification of annexin+ WT (blue) or SHARPIN-KO (red) B16F1s cultured with TNF. *P < 0.05 (n = 3/group; representative of 2 independent experiments). Immunoblot of WT and SHARPIN-KO B16F1 phenotype (inset, representative of 2 independent experiments). (B) Volumes of WT (closed circles) or SHARPIN-KO (open circles) B16F1 tumors coinjected into WT (red) or Rag–/– (black) hosts (n=7/group). **P < 0.05 at day 17 between noted conditions; *P < 0.05 versus SHARPIN-KO→WT host; *P <0.05 versus WT→WT host). (C) Tumor volume kinetics of WT (closed circles) or SHARPIN-KO (open circles) B16F1 cells coinjected into Tnf–/– hosts (n = 7/group). (D) YOYO3+ counts of WT (blue) or SHARPIN-KO (red) B16F1-tOVA–expressing target cells cocultured with effector OT-I T cells in the presence of control IgG (square, solid line) or anti-TNF (circle, dashed line) mAb at indicated E:T ratios (n = 3/group). *P < 0.05 versus WT + IgG; +P < 0.05 versus Shp + αTNF; #P < 0.05 versus WT + αTNF; **P < 0.05 WT + IgG versus WT + αTNF). (E) Tumor volume kinetics of WT (black lines) or SHARPIN-KO (red lines) B16F1 coinjected into WT hosts and treated with anti-PD1 (open circles) or IgG control (closed circles). Arrows indicate days of injection. (n = 7 for αPD1 therapy and n = 3–5 for IgG control). *P < 0.05 versus WT + αPD1; +P < 0.05 versus SHARPIN + αPD1. (F) Kinetic of difference in tumor size between WT and SHARPIN-KO B16F1 tumors in anti-PD1–treated hosts (n = 7). *P < 0.05 versus day 20. (G) Paired comparison of tumor size at day 20 after inoculation of IgG-treated (upper panel) or anti-PD1–treated (lower panel) hosts (n = 7 for αPD1 therapy and n = 3 for IgG control). *P < 0.05 versus WT condition. For all panels, statistics calculated between noted conditions by t test.