Targeting intracellular LMP2 with costimulatory signal–armed antibody-like TCR T cells
Jiali Cheng, Xuelian Hu, Zhenyu Dai, Yuhao Zeng, Jin Jin, Wei Mu, Qiaoe Wei, Xiangyin Jia, Jianwei Liu, Meng Xie, Qian Luo, Guang Hu, Gaoxiang Wang, Xiaojian Zhu, Jianfeng Zhou, Min Xiao, Jue Wang, Taochao Tan, Liang Huang
Jiali Cheng, Xuelian Hu, Zhenyu Dai, Yuhao Zeng, Jin Jin, Wei Mu, Qiaoe Wei, Xiangyin Jia, Jianwei Liu, Meng Xie, Qian Luo, Guang Hu, Gaoxiang Wang, Xiaojian Zhu, Jianfeng Zhou, Min Xiao, Jue Wang, Taochao Tan, Liang Huang
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Research Article Infectious disease Therapeutics

Targeting intracellular LMP2 with costimulatory signal–armed antibody-like TCR T cells

Abstract

Expanding the repertoire of CAR therapies to include intracellular antigens holds promise for treating a broad spectrum of malignancies. TCR-like T cells, capable of recognizing intracellular antigen–derived peptides in complex with HLA molecules (pHLA), represent a promising strategy in the field of engineered cellular therapy. This study introduced antibody-like TCR (abTCR) T cells that specifically targeted HLA-A*02:01–restricted LMP2426 peptides, a typical Epstein-Barr virus (EBV) latency II protein, for the treatment of EBV-associated lymphoproliferative diseases (EBV-LPDs). Compared with classic CAR T cells targeting the same epitope, abTCR T cells demonstrated superior efficiency, including increased CD107A expression, enhanced cytotoxicity, and elevated IFN-γ secretion, even when engaging with target cells that naturally present antigens. Moreover, a costimulatory signal–armed abTCR (Co-abTCR), which integrated a costimulatory structure with the abTCR, further enhanced the proliferation and in vivo tumoricidal efficacy of transfected T cells. Collectively, our study developed a potentially novel TCR-like T cell therapy that targets HLA-A*02/LMP2426 for the treatment of EBV-LPDs, providing a potential therapeutic solution for targeting of intracellular antigens in cancer immunotherapy.

Authors

Jiali Cheng, Xuelian Hu, Zhenyu Dai, Yuhao Zeng, Jin Jin, Wei Mu, Qiaoe Wei, Xiangyin Jia, Jianwei Liu, Meng Xie, Qian Luo, Guang Hu, Gaoxiang Wang, Xiaojian Zhu, Jianfeng Zhou, Min Xiao, Jue Wang, Taochao Tan, Liang Huang

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Figure 1

Screening and verification of A*02/LMP2-specific abTCR T cells.

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Screening and verification of A*02/LMP2-specific abTCR T cells. (A) Sche...
(A) Schematic of the screening and selection process. (B) Schematic diagram of abTCR and its functional format abTCR/CD3 complex. VH/VL, the variable domain of the heavy chain/light chain obtained from candidate phage clones; IgG CH and IgG CL, the constant domain 1 of the heavy chain and light chain of IgG; γ/δ, the transmembrane and intracellular domains of the γ/δ chain of TCR. Engineered abTCR consists of the 2 chains, VH-IgG CH-γ and VL-IgG CL-δ, which assemble and form complex with endogenous CD3 on T cell membrane to exert functions. (C) Cytolysis of 61abTCR T cells (red line) and mock T cells (gray line) against target-positive tumor cells, including Jeko1-LMP2 (LMP2426 overexpressed by PresentER minigene system), YT-A*02:01 (HLA-A*02:01 exogenously expressed), JVM2, and SNK6, as well as target-negative tumor cells, including Jeko1, YT, Raji, and THP1, at the specified E/T ratio for approximately 24 hours. Results from 3 or 4 replicates, with mean ± SD shown. Student’s t test was used for the statistical analysis. (D) CD107A expression on CD8+abTCR+ cells after overnight coincubation with cell lines of different tissue origins (summarized in Supplemental Table 1). The HLA-A*02 phenotype and LMP2 expression was indicated at the bottom of the figure. +, positive; ++, overexpressed; –, negative. The mean ± SD from 2–4 different donors are shown. Student’s t test was used for the statistical analysis. (E) CD107A expression on CD8+abTCR+ cells (red column) and CD8+CD3+ mock T cells (green column) after overnight incubation with healthy donor PBMC, including A*02+ and A*02–, and EBV-infected (copy number of EBV per 2 × 105 cells greater than 1 × 105) donor-derived PBMC, including A*02+ and A*02–. Jeko1-LMP2 or T2, pulsed with 0.5 μg/mL LMP2426, and buffer were set as positive and negative controls. Jeko1-LMP2–pulsed cells were used as the positive control, and buffer-treated cells served as the negative control. Sample size: 3–4. Each sample was tested once. Paired t test was used. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.