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Engineering TCR-preserved allogeneic CAR-T cells: leveraging cancer immune evasion principles to establish T-cell tolerance

Cellular & Molecular Immunology volume 22, pages 1285–1286 (2025)Cite this article

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Allogeneic “off-the-shelf” chimeric antigen receptor (CAR) T-cell therapy offers numerous advantages over autologous approaches. However, two critical challenges are encountered: graft-versus-host disease (GVHD) and poor persistence due to both extrinsic (host immune rejection) and intrinsic (TCR deletion-mediated) mechanisms. To prevent GVHD and prevent host-mediated graft rejection, allogeneic CAR-T-cell strategies typically employ selective editing of surface receptors or ligands, including (1) reducing the risk of GVHD by gene editing with TCR deletion [1, 2]; (2) diminishing allogeneic immunogenicity by knocking out HLA and β2 m, which can alleviate T-cell-mediated immune rejection but inadvertently activates NK cells [3]; and (3) preventing NK-cell-dependent lysis by overexpressing NK-inhibitory ligands, such as the nonclassical HLA molecules HLA-E and HLA-G [4]. Recently, Wu et al. introduced a novel strategy that targets signal peptide peptidase-like 3 (SPPL3) in allogeneic CAR-T cells. This approach confers dual resistance to both allogeneic rejection (mediated by T/NK cells) and Fas-dependent activation-induced cell death (AICD). Importantly, endogenous TCR expression was preserved, thereby enhancing CAR-T-cell persistence without increasing the risk of GVHD. In a phase 1 clinical trial, SPPL3-null CAR-T cells exhibited both safety and promising efficacy in patients with B-cell hematologic malignancies [5].

TCR deletion highlights a fundamental paradox in allogeneic CAR-T cells. While TCR ablation effectively prevents GVHD, it simultaneously impairs the survival of T cells, potentially through the loss of tonic TCR signaling [10]. Wang et al. revealed that in patients receiving TCRko CAR-T cells containing <1% residual TCR+ cells, >90% of surviving CAR-T cells reverted to TCR+ after two months [11]. In another study by Qasim et al., the initially infused cells comprised <0.7% of the TCR+ cells. However, almost all expanded CAR-T cells had transformed into TCR+ cells by day 56 [12]. In this study, CD3−CAR+ cells failed to persist beyond four weeks, yet residual CD3+CAR+ T cells in more than half of the patients underwent significant expansion [5]. These findings strongly suggest that TCR deletion diminishes allogeneic T-cell persistence. This hypothesis was further supported by murine competitive transfer experiments demonstrating the survival disadvantage of TCRKO T cells compared with TCR-preserved T cells. Notably, although these expanding cells exceeded the theoretical GVHD risk thresholds, no classical GVHD manifestations were observed. A key mechanism involves SPPL3 deletion-mediated alteration of TCR glycan modifications, which reduces graft-versus-host activity. This compelling outcome motivated a subsequent clinical trial evaluating TCR-preserved SPPL3KO/anti-CD19 CAR-T cells. Remarkably, despite HLA mismatch and no GVHD prophylaxis, even at cell doses sufficient to trigger fatal GVHD, none of the three patients developed acute or chronic GVHD. A self-limiting grade 1–3 rash observed in one case lacked histopathological hallmarks of GVHD (e.g., keratinocyte apoptosis or lichenoid inflammation) and resolved spontaneously without immunosuppressive therapy [5]. The underlying mechanism involves glycan-mediated attenuation of TCR signaling intensity, which may intrinsically limit GVHD development.

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Authors and Affiliations

  1. State Key Laboratory of Immune Response and Immunotherapy, Institute of Immunology, Biomedical Sciences and Health Laboratory of Anhui Province, School of Basic Medical Sciences, University of Science and Technology of China, Hefei, China

    Xiaodong Zheng & Zhigang Tian

  2. Center for Advanced Interdisciplinary Science and Biomedicine of IHM, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China

    Zhigang Tian

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Zheng, X., Tian, Z. Engineering TCR-preserved allogeneic CAR-T cells: leveraging cancer immune evasion principles to establish T-cell tolerance. Cell Mol Immunol 22, 1285–1286 (2025). https://doi.org/10.1038/s41423-025-01341-3

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