Department of Immunology, Charles University, 2nd Faculty of Medicine and University Hospital Motol, Prague, Czech Republic; Sotio, Prague, Czech Republic.
Sotio, Prague, Czech Republic.
Methods Enzymol. 2020;631:277-287. doi: 10.1016/bs.mie.2019.05.034. Epub 2019 Jun 27.
Natural killer (NK) cells constitute the predominant innate lymphocyte subset that mediates the anti-viral and anti-tumor immune responses. NK cells use an array of innate receptors to sense their environment and to respond to infections, cellular stress and transformation. The resulting NK cell activation, including cytotoxicity and cytokine production, is a fundamental component of the early immune response. The most recent discoveries in NK cell biology have stimulated the translational research that has led to remarkable results for the treatment of human malignancies. Therefore, the rapid isolation of NK cells from the peripheral blood or tumor microenvironment and the subsequent assessment of cytolytic function are crucial to the study of their potency and NK cell-mediated immunosurveillance. Here, we provide protocols for NK cell isolation and the assessment of NK cell cytotoxicity using flow cytometry.
自然杀伤 (NK) 细胞构成了介导抗病毒和抗肿瘤免疫反应的主要先天淋巴细胞亚群。NK 细胞利用一系列先天受体来感知其环境,并对感染、细胞应激和转化做出反应。由此产生的 NK 细胞激活,包括细胞毒性和细胞因子产生,是早期免疫反应的一个基本组成部分。NK 细胞生物学的最新发现激发了转化研究,这些研究为治疗人类恶性肿瘤带来了显著的成果。因此,从外周血或肿瘤微环境中快速分离 NK 细胞,并随后评估其细胞毒性功能,对于研究其效力和 NK 细胞介导的免疫监视至关重要。在这里,我们提供了使用流式细胞术分离 NK 细胞和评估 NK 细胞细胞毒性的方案。