Department of Biological and Environmental Sciences, Longwood University, Farmville, VA, USA.
Methods Mol Biol. 2021;2265:645-654. doi: 10.1007/978-1-0716-1205-7_44.
Expression of chimeric antigen receptors can redirect T cell specificity and allow for MHC-independent recognition of melanoma associated antigens. Retroviral transduction is used to express chimeric antigen receptor constructs in murine CD4+ and CD8+ T cells. Here we describe the production of retroviral supernatants and the activation, transduction, expansion, and selection of murine T cells expressing the chimeric-PD1 receptor. This protocol can be modified for any murine chimeric antigen receptor construct.
嵌合抗原受体的表达可以重新定向 T 细胞的特异性,并允许 MHC 非依赖性识别黑色素瘤相关抗原。逆转录病毒转导用于在小鼠 CD4+和 CD8+T 细胞中表达嵌合抗原受体构建体。在这里,我们描述了产生逆转录病毒上清液以及激活、转导、扩增和选择表达嵌合 PD1 受体的小鼠 T 细胞的方法。该方案可针对任何小鼠嵌合抗原受体构建体进行修改。