A CD4+ T lymphocyte–specific TCR/GSDMD/IL-2 axis facilitates antitumor immunity
Yihan Yao, … , Wanqiang Sheng, Xiaojian Wang
Yihan Yao, … , Wanqiang Sheng, Xiaojian Wang
Published August 1, 2025
Citation Information: J Clin Invest. 2025;135(15):e191119. https://doi.org/10.1172/JCI191119.
View: Text | PDF
Research Article Immunology Oncology

A CD4+ T lymphocyte–specific TCR/GSDMD/IL-2 axis facilitates antitumor immunity

Abstract

Gasdermin (GSDM) family proteins mediate tumor pyroptosis and impact cancer progression, but other than that, their involvement in the tumor immune microenvironment remains largely unknown. Here, we show that activation of GSDMD in human tumor specimens mainly occurs in tumor-infiltrating leukocytes. Significantly, GSDMD deficiency or its inactivation in CD4+ T cells disabled CD8+ T cell–mediated antitumor immunity and caused tumor outgrowth in mice. Further study uncovered that, via inducing IL-2 production, GSDMD was required for CD4+ T cells to provide help to CD8+ T cell function. Mechanistically, GSDMD was cleaved by TCR stimulation–activated caspase-8 to form GSDMD-N pores, which enhanced Ca2+ influx for IL-2 induction. Moreover, GSDMD activation and function were conserved in human CD4+ T cells and associated with favorable prognosis and improved response to anti–PD-1 immunotherapy in colonic and pancreatic cancer. We believe this study identifies a new nonpyroptotic role of GSDMD in tumor immunity, proposing GSDMD as a potential target for cancer immunotherapy.

Authors

Yihan Yao, Lingling Wang, Weiqin Jiang, Ning Wang, Mengjie Li, Wenlong Lin, Ting Zhang, Wanqiang Sheng, Xiaojian Wang

×

Figure 7

GSDMD activation in CD4+ T cells correlates with survival benefit and immunotherapy efficacy in colorectal cancer and pancreatic adenocarcinoma.

Options: View larger image (or click on image) Download as PowerPoint
GSDMD activation in CD4+ T cells correlates with survival benefit and im...
(A) Immunoblot analysis of GSDMD protein activation probed with a mixture of full-length and N-fragment–specific GSDMD antibodies in human CD4+ T cells isolated from PBMCs and activated by anti-hCD3/hCD28 for the indicated times. Red asterisks indicate cleaved GSDMD fragments. (B and C) Flow cytometry analysis of percentages of IL-2–expressing human CD4+ T cells (B) and IL-2 MFI in human CD4+ T cells (C) activated in vitro for 24 hours in the presence or absence of DSF (n = 3 per group). (D and E) Percentages of IL-2–expressing cells within shGSDMD-GFP–transduced (GFP+) or nontransduced (GFP–) human CD4+ T cells (D) and IL-2 MFI of GFP– and GFP+ human CD4+ T cells (E). (F–H) Immunofluorescence staining for GSDMD-N (red) and CD4 (green) in tumor tissues from patients with colorectal cancer (F) or pancreatic cancer (G). Percentages of GSDMD-N+ cells among CD4+ cells were quantified from 5 independent fields of view within colorectal cancer (CRC) and pancreatic adenocarcinoma (PAAD) (H). Scale bars: 10 μm. White arrowheads indicate GSDMD-N– and CD4-coexpressing cells. (I and J) Patients with colorectal cancer (I) or pancreatic cancer (J) were stratified into high-activation and low-activation groups based on the median ratio of GSDMD-active CD4+ TILs to total CD4+ TILs, and Kaplan-Meier curves of overall survival were calculated. (K) Tumor growth curves (left) and tumor weights (right) of MC38 tumors implanted in Gsdmdfl/fl and Gsdmdfl/fl CD4cre mice treated with anti–PD-1. (L) Left: Percentages of GSDMD-N+ cells in tumor-infiltrating CD4+ T cells in immunotherapy-responsive and –nonresponsive colorectal cancer patients. Right: Representative images of GSDMD activation in tumor-infiltrating CD4+ T cells. Data are presented as mean ± SEM (B–E and H–L) and are representative of at least 2 independent experiments (B–H and K). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, as determined by unpaired 2-tailed Student’s t tests (B, C, and L), paired 2-tailed Student’s t test (D and E), 1-way ANOVA for tumor weight (K), or log-rank test (I and J).