Calderon Hugo, Mamonkin Maksim, Guedan Sonia
Department of Hematology and Oncology, Hospital Clinic, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain.
Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, TX, USA.
Methods Mol Biol. 2020;2086:223-236. doi: 10.1007/978-1-0716-0146-4_17.
CARs are synthetic receptors designed to drive antigen-specific activation upon binding of the scFv to its cognate antigen. However, CARs can also elicit different levels of ligand-independent constitutive signaling, also known as tonic signaling. Chronic T cell activation is observed in certain combinations of scFv, hinge, and costimulatory domains and may be increased due to high levels of CAR expression. Tonic signaling can be identified during primary T cell expansion due to differences in the phenotype and growth of CAR-T cells compared to control T cells. CARs displaying tonic signaling are associated with accelerated T cell differentiation and exhaustion and impaired antitumor effects. Selecting CARs which configuration does not induce tonic signaling is important to enhance antigen-specific T cell responses. In this chapter, we describe in detail different protocols to identify tonic signaling driven by CARs during primary T cell ex vivo expansion.
嵌合抗原受体（CAR）是一种合成受体，设计用于在单链抗体片段（scFv）与其同源抗原结合时驱动抗原特异性激活。然而，CAR也可引发不同水平的不依赖配体的组成性信号传导，也称为张力信号传导。在scFv、铰链区和共刺激结构域的某些组合中观察到慢性T细胞激活，并且由于CAR的高表达水平，这种激活可能会增加。由于与对照T细胞相比，CAR-T细胞在表型和生长方面存在差异，因此在原发性T细胞扩增过程中可以识别张力信号传导。显示张力信号传导的CAR与T细胞分化加速、耗竭以及抗肿瘤作用受损有关。选择不诱导张力信号传导的CAR构型对于增强抗原特异性T细胞反应很重要。在本章中，我们详细描述了在原发性T细胞体外扩增过程中识别由CAR驱动的张力信号传导的不同方案。
