Abstract
Tumor infiltrating T cells (TIL) are key players in the anti-tumor immune response. However, chronic exposure to tumor-derived antigens drives the differentiation into ‘exhausted’ TILs. Whether intratumoral dendritic cells (DC) can mitigate TILs exhaustion and maintain function is unclear. Here, we develop a bispecific DC-T cell engager (BiDT), consisting of an anti-TIM3-IFN fusion protein, and demonstrate that, in preclinical mouse tumor models, this engager simultaneously targets TIM3 on exhausted TILs and activates DCs via the IFNAR receptor. Mechanistically, BiDT reactivates exhausted TIM3+TILs by preventing apoptosis through increased Bcl-2 expression and enhances DC function to reactivate T cells via IL-2 signalling and co-stimulatory CD80/86-CD28 interactions within the tumor microenvironment. Finally, to mitigate IFNα-induced toxicity, we engineer a Pro-BiDT engager featuring a pro-IFNα and report potent antitumor activity with reduced systemic toxicity. Thus, by bridging DC-T cells together, BiDT treatment enhances the critical communication pathways and cellular circuits necessary for effective anti-tumor immunity.
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All data are included in the Supplementary Information or available from the authors, as are unique reagents used in this Article. The raw numbers for charts and graphs are available in the Source Data file whenever possible. The single-cell RNA-seq data have been deposited in Gene Expression Omnibus under the accession number GSE320312. Correspondence and requests for materials should be addressed to Yang-Xin Fu (yangxinfu@tsinghua.edu.cn), Wenyan Wang (wywang2022@tsinghua.edu.cn) or Xuhao Zhang (xuhaozhang@cqmu.edu.cn). Source data are provided with this paper.
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Acknowledgements
We are grateful to Hua Peng (Institute of Biophysics, Chinese Academy of Sciences, Beijing) for Ifnar1−/− and Ifnar1fl/fl mice. Li Wu (Tsinghua University, Beijing) for Zbtb46-Cre mice; Gencheng Han (Beijing Institute of Basic Medical Sciences, Beijing) for Lyz2-Cre and Havcr2 fl/fl mice; Xiaohuan Guo (Tsinghua University, Beijing) for Cd4-Cre mice. We thank Mingwan Tang, Yutong Fu, Weian Cao, and Li Yu for providing experiment materials and helpful discussions. We thank the faculty in the animal facility of Tsinghua University. This work was supported by grants from the National Natural Science Foundation of China (82250710684, to Y-X.F., 32370967 to W.W., 32300773 to X.S.) and International Postdoctoral Exchange Fellowship Program (to X. Zhang).
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Conceptualization: X.Z., Y.G., W.H. and Y-X. F Methodology: X.Z., Y.G., W.H., Y.L., X.Y., X.S., H.L., H.L., X.Y., Y.G. Investigation: X.Z., Y.G., W.H., W.W., M.Z., H.P. Funding acquisition: X.Z., W.W., and Y-X.F. Supervision: W.W., Y-X. F. Writing – original draft: X.Z., Y.G., W.H. Writing – review & editing: X.Z., Y.G., W.H. W.W. and Y-X.F.
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Zhang, X., Gao, Y., Hu, W. et al. Reactivating exhausted tumor-infiltrating T cells by a bispecific DC-T cell engager in mice. Nat Commun (2026). https://doi.org/10.1038/s41467-026-70876-4
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DOI: https://doi.org/10.1038/s41467-026-70876-4
