减弱初始刺激助力过继T细胞克服抗原逃逸

李义然; 潘俊成; 施小山

doi:10.12211/2096-8280.2026-018

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减弱初始刺激助力过继T细胞克服抗原逃逸

研究论文 | 更新时间：2026-05-12

- 减弱初始刺激助力过继T细胞克服抗原逃逸
- Attenuated initial stimulation confers adoptive T cell potency against antigen mutational escape
- 合成生物学 2026年页码：1-14
- 作者机构：
  
  1.中国科学院深圳先进技术研究院，合成生物学研究所，广东深圳 518055
  2.中国科学院大学，北京 100049
- 作者简介：
  
  李义然（2000—），男，硕士研究生。研究方向为新型T细胞合成免疫学改造策略研发。
  施小山（1987—），男，博士，研究员。研究方向为免疫细胞信号机制研究与新型合成免疫学改造策略研发。
- 基金信息：
  
  国家重点研发计划(2023YFA0915700);深圳高层次人才团队(KQTD20240729102054070)
- DOI：10.12211/2096-8280.2026-018
  中图分类号： Q813
- 收稿：2026-03-26，
  
  修回：2026-05-10，
  
  网络首发：2026-05-12，
- 稿件说明：
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李义然，潘俊成，施小山. 减弱初始刺激助力过继T细胞克服抗原逃逸［J］. 合成生物学， 2026， 7. DOI： 10.12211/2096-8280.2026-018

LI Yiran， PAN Juncheng， SHI Xiaoshan. Attenuated initial stimulation confers adoptive T cell potency against antigen mutational escape［J］. Synthetic Biology Journal， 2026， 7. DOI： 10.12211/2096-8280.2026-018
李义然，潘俊成，施小山. 减弱初始刺激助力过继T细胞克服抗原逃逸［J］. 合成生物学， 2026， 7. DOI： 10.12211/2096-8280.2026-018 DOI：

LI Yiran， PAN Juncheng， SHI Xiaoshan. Attenuated initial stimulation confers adoptive T cell potency against antigen mutational escape［J］. Synthetic Biology Journal， 2026， 7. DOI： 10.12211/2096-8280.2026-018 DOI：

摘要

过继性T细胞疗法在肿瘤治疗中展现出巨大潜力，但仍面临持续高强度肿瘤抗原刺激引起的T细胞耗竭，及肿瘤抗原低亲和力突变导致的免疫逃逸问题。传统的过继性T细胞生产工艺长期以来追求最大化T细胞受体（TCR）刺激强度以增强T细胞活化与扩增的策略，但该策略往往加速终末T细胞分化并导致不可逆的耗竭，同时未能有效应对抗原突变逃逸的问题。本研究利用常用的小鼠OT-1 T细胞及人原代T细胞模型，包括识别实体瘤A375细胞的1G4 TCR-T细胞，系统探究了初始T细胞受体刺激强度在体外制备过程中对过继性T细胞状态及效应功能的影响。结果表明，通过降低抗原肽浓度、采用低亲和力抗原或降低CD3/CD28抗体磁珠用量以适度减弱刺激强度，可在不减弱T细胞充分活化与扩增的前提下，显著降低其PD-1、LAG3等抑制性受体的表达。在功能层面，弱刺激强度制备的T细胞不仅在应对肿瘤细胞时分泌更多的TNF-α、启动更迅速的杀伤效应，更对表达低亲和力突变抗原肿瘤展现出更好的控制能力。综上，本研究揭示了适度减弱体外制备期初始刺激强度可显著提升过继性T细胞的效能并有效应对肿瘤抗原突变逃逸，为过继性T细胞制备的工艺优化提供了一种易借鉴、低成本且潜在普适的新思路。

Abstract

Adoptive T cell (ACT) therapy has revolutionized cancer immunotherapy with its targeted and durable anti-tumor potency

yet its broad clinical success

particularly against solid tumors

is severely compromised by two critical barriers: T cell exhaustion driven by persistent high-level T cell receptor (TCR) stimul

ation

and tumor immune escape via low-affinity mutations in tumor antigens. Conventional ACT manufacturing has long been guided by the paradigm of maximizing TCR stimulation strength to boost T cell activation and expansion

but this strategy frequently accelerates terminal T cell differentiation and irreversible exhaustion

while failing to address antigen mutational escape

creating an urgent unmet need for optimized

clinically translatable manufacturing protocols. In this study

we systematically investigated how initial TCR stimulation strength during in vitro manufacturing shapes the phenotypic state and anti-tumor effector function of T cells

using the well-established murine OT-1 CD8

T cell model and clinically relevant human primary T cells including the 1G4 TCR-T system for solid tumor A375 cell targeting

and attenuated TCR stimulation via three readily translatable approaches: titrating down cognate antigen peptide concentration

using low-affinity antigen variants

and reducing the dosage of CD3/CD28 antibody-coated stimulatory beads. Our results demonstrated that moderately attenuated initial stimulation supported robust T cell activation and expansion

while significantly downregulating the expression of exhaustion-associated inhibitory receptors including PD-1

LAG3 and TIGIT. Functionally

weakly stimulated T cells exhibited faster cytotoxic kinetics against tumor cells

higher TNF-α secretion

and most notably

superior and sustained control of tumors expressing low-affinity mutant antigens both in vitro across multiple effector-to-target ratios and in an in vivo immunocompromised mouse tumor model

with these core findings fully recapitulated in human primary T cells and the 1G4 TCR-T system where attenuated stimulation enhanced antigen-specific cytotoxicity and proliferative capacity. Collectively

this study reveals that fine-tuning and moderately reducing initial TCR stimulation strength during in vitro manufacturing could serve as a facile

cost-effective and potential univers

al strategy to generate high-potency T cells for ACT

which mitigates T cell exhaustion and effectively counteracts antigen mutational escape

providing a practical and translatable rationale to optimize ACT manufacturing protocols and improve clinical outcomes for patients with advanced malignancies.

关键词

Keywords

references

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