Cheadle Eleanor J, Sheard Vicky, Hombach Andreas A, Chmielewski Markus, Riet Tobias, Berrevoets Cor, Schooten Erik, Lamers Cor, Abken Hinrich, Debets Reno, Gilham David E
Cell Therapy Group, Department of Medical Oncology, School of Cancer and Enabling Sciences, Manchester Academic Health Science Center, The University of Manchester, Manchester, UK.
Methods Mol Biol. 2012;907:645-66. doi: 10.1007/978-1-61779-974-7_36.
The Chimeric Antigen Receptor (CAR) consists of an antibody-derived targeting domain fused with T-cell signaling domains that, when expressed by a T-cell, endows the T-cell with antigen specificity determined by the targeting domain of the CAR. CARs can potentially redirect the effector functions of a T-cell towards any protein and nonprotein target expressed on the cell surface as long as an antibody or similar targeting domain is available. This strategy thereby avoids the requirement of antigen processing and presentation by the target cell and is applicable to nonclassical T-cell targets like carbohydrates. This circumvention of HLA-restriction means that the CAR T-cell approach can be used as a generic tool broadening the potential of applicability of adoptive T-cell therapy. Proof-of-principle studies focusing upon the investigation of the potency of CAR T-cells have primarily focused upon the genetic modification of human and mouse T-cells for therapy. This chapter focuses upon methods to modify T-cells from both species to generate CAR T-cells for functional testing.
嵌合抗原受体（CAR）由与T细胞信号结构域融合的抗体衍生靶向结构域组成，当由T细胞表达时，该结构域赋予T细胞由CAR靶向结构域决定的抗原特异性。只要有抗体或类似的靶向结构域，CAR就有可能将T细胞的效应功能重定向到细胞表面表达的任何蛋白质和非蛋白质靶标上。因此，该策略避免了靶细胞对抗原的加工和呈递的要求，并且适用于像碳水化合物这样的非经典T细胞靶标。这种对HLA限制的规避意味着CAR T细胞方法可以用作拓宽过继性T细胞疗法应用潜力的通用工具。专注于研究CAR T细胞效力的原理验证研究主要集中在对人和小鼠T细胞进行基因改造以用于治疗。本章重点介绍从这两个物种中修饰T细胞以产生用于功能测试的CAR T细胞的方法。
