Chimeric antigen receptor (CAR) is a hybrid molecule consisting of an antigen-binding domain and a signal transduction domain. The artificial T cells expressing CAR (CAR-T cells) are expected to be a useful tool for treatment of various diseases, such as cancer. The addition of a co-stimulatory signal domain (CSSD) to CAR is shown to be critical for modulating CAR-T cell activities. However, the interplay among types of CSSDs, effector functions, and characteristics of CAR-T cells is largely unknown. To elucidate the interplay, we analyzed effector functions, differentiation to memory T cell subsets, exhaustion, and energy metabolism of the CAR-T cells with different CSSDs. Comparing to the CAR-T cells bearing a CD28- or 4-1BB-derived CSSD, which are currently used for CAR-T cell development, we found that the CAR-T cells with a herpes virus entry mediator (HVEM)-derived CSSD exhibited enhanced effector functions and efficient and balanced differentiation to both central and effector memory subsets, associated with an elevated energy metabolism and a reduced level of exhaustion. Thus, we developed the CAR-T cells bearing the CSSD derived from HVEM with high functional potency. The HVEM-derived CSSD may be useful for developing effective CAR-T cells.

嵌合抗原受体（CAR）是由抗原结合结构域和信号转导结构域组成的杂合分子。预期表达CAR的人工T细胞（CAR-T细胞）将成为治疗各种疾病（例如癌症）的有用工具。已显示向CAR添加共刺激信号域（CSSD）对于调节CAR-T细胞活性至关重要。然而，CSSD类型，效应子功能和CAR-T细胞特征之间的相互作用尚不清楚。为了阐明相互作用，我们分析了效应器功能，具有不同CSSD的CAR-T细胞向记忆T细胞亚群的分化，衰竭和能量代谢。与目前用于CAR-T细胞发育的带有CD28或4-1BB的CSSD的CAR-T细胞相比，我们发现带有疱疹病毒进入介体（HVEM）的CSSD的CAR-T细胞表现出增强的效应子功能以及对中央和效应子记忆子集的有效且平衡的分化，与能量代谢升高和疲惫程度降低相关。因此，我们开发了具有来自HVEM的具有高功能效力的CSSD的CAR-T细胞。HVEM衍生的CSSD可用于开发有效的CAR-T细胞。