Shi Guilan, Heller Richard
Department of Medical Engineering, Colleges of Medicine and Engineering, University of South Florida, Tampa, FL, USA.
Methods Mol Biol. 2021;2265:111-118. doi: 10.1007/978-1-0716-1205-7_8.
Within the adaptive and innate immune system, effector lymphocytes known as cytotoxic T cells (CTLs) or natural killer (NK) cells play an essential role in host defense against tumor cells and virally infected cells. Here we describe a flow cytometry-based method to quantify CTLs or NK cell cytotoxic activity against melanoma cells. In this assay, spleen cells, peripheral blood mononuclear cells (PBMCs), or purified NK cell preparations are co-incubated at different ratios with a target tumor cell line. The target cells are pre-labeled with a fluorescent dye to allow their discrimination from the effector cells. After the incubation period, killed target cells are identified by a nucleic acid stain, which specifically permeates dead cells. This method is amenable to both diagnostic and research applications.
在适应性和先天性免疫系统中，被称为细胞毒性T细胞（CTL）或自然杀伤（NK）细胞的效应淋巴细胞在宿主抵御肿瘤细胞和病毒感染细胞的过程中发挥着至关重要的作用。在此，我们描述一种基于流式细胞术的方法，用于量化CTL或NK细胞对黑色素瘤细胞的细胞毒性活性。在该检测中，脾细胞、外周血单核细胞（PBMC）或纯化的NK细胞制剂以不同比例与靶肿瘤细胞系共同孵育。靶细胞预先用荧光染料标记，以便将它们与效应细胞区分开来。孵育期过后，通过一种特异性穿透死细胞的核酸染色来识别被杀死的靶细胞。该方法适用于诊断和研究应用。
