Almåsbak Hilde, Sioud Mouldy, Rasmussen Anne-Marie
Cellular Medicine, Bioproduction, Thermo Fisher Scientific, Oslo, Norway.
Division of Cancer Medicine, Department of Cancer Immunology, Institute for Cancer Research, Oslo University Hospital Radiumhospitalet, Oslo, Norway.
Methods Mol Biol. 2025;2965:285-321. doi: 10.1007/978-1-0716-4742-4_14.
Adoptive T-cell therapy is a promising cancer immunotherapy, but isolating tumor-specific cytotoxic T cells is time-consuming and often unsuccessful. An alternative approach involves genetically modifying T cells to express tumor antigen-specific T-cell receptors (TCRs) or chimeric antigen receptors (CARs), enabling the redirection of large numbers of immune cells to target malignant cells. CARs combine the specificity of antibody-derived single-chain variable fragments with T-cell signaling domains (e.g., CD3ζ, CD28, 4-1BB), enabling T cells to recognize and kill antigen-positive cancer cells in an MHC-independent manner. This chapter outlines protocols for large-scale T-cell expansion and medium-scale production of messenger RNA (mRNA) CAR-T cells using electroporation as a delivery method. Two CARs are used as model systems: one targeting CD19 and another targeting an antigen expressed on hematological malignancies and solid tumors.
过继性T细胞疗法是一种很有前景的癌症免疫疗法，但分离肿瘤特异性细胞毒性T细胞既耗时又常常不成功。另一种方法是对T细胞进行基因改造，使其表达肿瘤抗原特异性T细胞受体（TCR）或嵌合抗原受体（CAR），从而使大量免疫细胞能够重新定向以靶向恶性细胞。CAR将抗体衍生的单链可变片段的特异性与T细胞信号结构域（如CD3ζ、CD28、4-1BB）结合在一起，使T细胞能够以不依赖MHC的方式识别并杀死抗原阳性癌细胞。本章概述了使用电穿孔作为递送方法进行大规模T细胞扩增和中规模信使核糖核酸（mRNA）CAR-T细胞生产的方案。使用两种CAR作为模型系统：一种靶向CD19，另一种靶向在血液系统恶性肿瘤和实体瘤上表达的一种抗原。
