Transcriptomic profile of CD8+ circulating tumor-reactive lymphocytes (CTRL) isolated by MHC multimers. Transcriptomic profile of CD8+ circulating tumor-reactive lymphocytes (CTRL) isolated by MHC multimers

Circulating tumor-reactive lymphocytes (CTRL) present in blood circulation at a very low frequency. We efficiently isolated them based on high-performance microfluidics and MHC multimer binding assay. Using an ultra-low input bulk RNAseq workflow, we systematically characterize the transcriptomic profile of tumor-reactive lymphocytes and identified novel biomarkers for multimer-free isolation. Two animal models were studied - B16 melanoma model and CT-26 colon cancer model. Overall design: CD8+ lymphocytes in PBMC were isolated based on their binding against tumor-reactive multimers. Isolated subpopulations were sequenced. Differentially expressed genes were compared.

循环肿瘤反应性淋巴细胞（Circulating Tumor-Reactive Lymphocytes, CTRL）在血液循环中仅以极低频率存在。本研究依托高性能微流控技术与MHC多聚体结合实验，实现了此类细胞的高效分离。我们采用了超低起始量批量RNA测序（RNA Sequencing, RNA-seq）流程，系统表征了肿瘤反应性淋巴细胞的转录组谱，并鉴定出可用于无多聚体分离的新型生物标志物。本研究共使用两种动物模型：B16黑色素瘤模型与CT-26结肠癌模型。实验设计：从外周血单个核细胞（Peripheral Blood Mononuclear Cell, PBMC）中分离CD8+淋巴细胞，基于其与肿瘤反应性多聚体的结合能力进行分选，对分离得到的细胞亚群进行测序，并比较各组的差异表达基因。