Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33614, USA.
Molecular Genomics Core Facility, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33614, USA.
STAR Protoc. 2022 Aug 26;3(3):101649. doi: 10.1016/j.xpro.2022.101649. eCollection 2022 Sep 16.
Understanding the heterogenicity of tumor-infiltraing lymphocyte (TIL) populations and the immunobiology in human cancer is a key to establish efficient immunotherapies. Here, we have established a protocol for the characterization of CD8 TILs in tumors by single-cell RNA-seq paired to VDJ profiling and chromatin structure including dissociation of tumor biopsies. We have also provided guidance for subsequent fluorescence-activated cell sorting (FACS), single-cell encapsulation, bioinformatics analysis, and troubleshooting. For complete details on the use and execution of this protocol, please refer to Anadon et al. (2022).
了解肿瘤浸润淋巴细胞 (TIL) 群体的异质性和人类癌症中的免疫生物学是建立有效免疫疗法的关键。在这里，我们建立了一种通过单细胞 RNA-seq 与 VDJ 分析以及包括肿瘤活检解离在内的染色质结构配对来表征肿瘤中 CD8 TIL 的方案。我们还为后续的荧光激活细胞分选 (FACS)、单细胞包封、生物信息学分析和故障排除提供了指导。有关该方案使用和执行的完整详细信息，请参阅 Anadon 等人 (2022)。
