Lorenzo-Herrero Seila, Sordo-Bahamonde Christian, González Segundo, López-Soto Alejandro
Departamento de Biología Funcional, Inmunología, Universidad of Oviedo, Oviedo, Spain.
Instituto Universitario de Oncología del Principado de Asturias (IUOPA), Instituto de Investigación Sanitaria del Principado de Asturias (IISPA), Oviedo, Spain.
Methods Mol Biol. 2019;1884:131-139. doi: 10.1007/978-1-4939-8885-3_8.
The immune system is able to detect and eliminate nascent and developing tumors. Thus, T lymphocytes of the adaptive immunity recognize cancer cells by detecting tumor-associated antigens, whereas certain innate immune cells scan for molecules that are mainly overexpressed on malignant and infected cells. The best example of the latter is natural killer (NK) cells. The activity of these immune cells is strictly governed by a balance between positive and negative signals provided by stress-regulated molecules that bind NK cell membrane receptors, such as the activatory receptor NKG2D. Given the key role of NK cells in eradicating cancer cells, the development of assays to study the cellular and molecular determinants of NK cell antitumor activity are of great relevance. In this chapter, we describe a flow cytometric in vitro assay to evaluate the cytotoxic activity of NK cells against a given target cell, including tumor cells. Moreover, this method is highly versatile, as it can be complemented with the use of antibodies blocking cell surface proteins (receptors or ligands) or a variety of chemical modulators, which allows the elucidation of molecules and signaling pathways that regulate NK cell anticancer function.
免疫系统能够检测并清除新生的和正在发展的肿瘤。因此，适应性免疫的T淋巴细胞通过检测肿瘤相关抗原来识别癌细胞，而某些固有免疫细胞则扫描主要在恶性细胞和受感染细胞上过度表达的分子。后者的最佳例子是自然杀伤（NK）细胞。这些免疫细胞的活性严格受与NK细胞膜受体结合的应激调节分子提供的正负信号之间平衡的支配，例如激活受体NKG2D。鉴于NK细胞在根除癌细胞中的关键作用，开发用于研究NK细胞抗肿瘤活性的细胞和分子决定因素的检测方法具有重要意义。在本章中，我们描述了一种流式细胞术体外检测方法，用于评估NK细胞对给定靶细胞（包括肿瘤细胞）的细胞毒性活性。此外，该方法具有高度的通用性，因为它可以与阻断细胞表面蛋白（受体或配体）的抗体或各种化学调节剂结合使用，这有助于阐明调节NK细胞抗癌功能的分子和信号通路。
