通过流式细胞术对人自然杀伤细胞进行功能分析。
Functional analysis of human NK cells by flow cytometry.
作者信息
Bryceson Yenan T, Fauriat Cyril, Nunes João M, Wood Stephanie M, Björkström Niklas K, Long Eric O, Ljunggren Hans-Gustaf
机构信息
Center for Infectious Medicine, Department of Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, Stockholm, Sweden.
出版信息
Methods Mol Biol. 2010;612:335-52. doi: 10.1007/978-1-60761-362-6_23.
Abstract
Natural killer (NK) cells are a subset of lymphocytes that contribute to innate immunity through cytokine secretion and target cell lysis. NK cell function is regulated by a multiplicity of activating and inhibitory receptors. The advance in instrumentation for multi-color flow cytometry and the generation of specific mAbs for different epitopes related to phenotypic and functional parameters have facilitated our understanding of NK cell responses. Here, we provide protocols for flow cytometric evaluation of degranulation and cytokine production by human NK cells from peripheral blood at the single-cell level. In addition to offering insight into the regulation of human NK cell responses, these techniques are applicable to the assessment of various clinical conditions, including the diagnosis of immunodeficiency syndromes.
摘要
自然杀伤（NK）细胞是淋巴细胞的一个亚群，通过细胞因子分泌和靶细胞裂解作用参与固有免疫。NK细胞功能受多种激活和抑制性受体调节。多色流式细胞术检测仪器的进步以及针对与表型和功能参数相关的不同表位产生的特异性单克隆抗体，有助于我们理解NK细胞反应。在此，我们提供了在单细胞水平上对外周血人NK细胞脱颗粒和细胞因子产生进行流式细胞术评估的方案。除了有助于深入了解人NK细胞反应的调节机制外，这些技术还可用于评估各种临床状况，包括免疫缺陷综合征的诊断。
相似文献
1
Functional analysis of human NK cells by flow cytometry.通过流式细胞术对人自然杀伤细胞进行功能分析。
Methods Mol Biol. 2010;612:335-52. doi: 10.1007/978-1-60761-362-6_23.
2
Flow Cytometry Analysis of NK Cell Phenotype and Function in Aging.衰老过程中自然杀伤细胞表型和功能的流式细胞术分析
Methods Mol Biol. 2015;1343:9-18. doi: 10.1007/978-1-4939-2963-4_2.
3
Analysis of the KIR repertoire in human NK cells by flow cytometry.通过流式细胞术分析人类自然杀伤细胞中的杀伤细胞免疫球蛋白样受体库。
Methods Mol Biol. 2010;612:353-64. doi: 10.1007/978-1-60761-362-6_24.
4
Optimization of Human NK Cell Manufacturing: Fully Automated Separation, Improved Ex Vivo Expansion Using IL-21 with Autologous Feeder Cells, and Generation of Anti-CD123-CAR-Expressing Effector Cells.人自然杀伤细胞制备的优化：全自动分离，使用 IL-21 和自体饲养细胞进行体外扩增的改进，以及生成表达抗 CD123-CAR 的效应细胞。
Hum Gene Ther. 2017 Oct;28(10):897-913. doi: 10.1089/hum.2017.157. Epub 2017 Aug 15.
5
A rapid method for assessment of natural killer cell function after multiple receptor crosslinking.一种快速评估多次受体交联后自然杀伤细胞功能的方法。
J Immunol Methods. 2011 Mar 7;366(1-2):52-9. doi: 10.1016/j.jim.2011.01.007. Epub 2011 Jan 28.
6
A cytofluorimetric assay to evaluate intracellular cytokine production by NK cells.一种用于评估 NK 细胞细胞内细胞因子产生的细胞荧光分析方法。
Methods Enzymol. 2020;631:343-355. doi: 10.1016/bs.mie.2019.05.049. Epub 2019 Jun 17.
7
Protocol for the clonal analysis of NK cell effector functions by multi-parameter flow cytometry.通过多参数流式细胞术进行自然杀伤细胞效应功能克隆分析的方案。
Methods Mol Biol. 2012;903:381-92. doi: 10.1007/978-1-61779-937-2_26.
8
Natural Killer Cells in Graves' Disease: Increased Frequency but Impaired Degranulation Ability Compared to Healthy Controls.格雷夫斯病中的自然杀伤细胞：与健康对照相比，频率增加但脱颗粒能力受损。
Int J Mol Sci. 2025 Jan 24;26(3):977. doi: 10.3390/ijms26030977.
9
Flow cytometry in detection of abnormalities of natural killer cell.流式细胞术检测自然杀伤细胞异常。
Adv Exp Med Biol. 2013;756:303-11. doi: 10.1007/978-94-007-4549-0_37.
10
Measurement of NK activity in whole blood by the CD69 up-regulation after co-incubation with K562, comparison with NK cytotoxicity assays and CD107a degranulation assay.通过与 K562 共孵育后 CD69 的上调来测量全血中的 NK 活性，与 NK 细胞毒性测定和 CD107a 脱颗粒测定进行比较。
J Immunol Methods. 2011 Sep 30;372(1-2):187-95. doi: 10.1016/j.jim.2011.07.016. Epub 2011 Aug 2.
引用本文的文献
1
Complete CD16A Deficiency and Defective NK Cell Function in a Man Living with HIV.一名HIV感染者的完全CD16A缺陷与自然杀伤细胞功能缺陷
J Clin Immunol. 2025 May 24;45(1):98. doi: 10.1007/s10875-025-01886-y.
2
ADCC: the rock band led by therapeutic antibodies, tumor and immune cells.抗体依赖的细胞介导的细胞毒性作用（ADCC）：由治疗性抗体、肿瘤和免疫细胞引领的摇滚乐队。
需注意，你提供的英文原文表述在医学语境下不太常规且易造成误解，正常的医学概念“ADCC”完整表述为“Antibody-dependent cell-mediated cytotoxicity” ，直译为“抗体依赖的细胞介导的细胞毒性作用” 。 以上译文是按照你要求对这段奇特表述进行的翻译 。
Front Immunol. 2025 Apr 16;16:1548292. doi: 10.3389/fimmu.2025.1548292. eCollection 2025.
3
Innate immune cells in acute and chronic kidney disease.急性和慢性肾病中的固有免疫细胞。
Nat Rev Nephrol. 2025 Apr 22. doi: 10.1038/s41581-025-00958-x.
4
Cytolytic γδ T-cells and IFNγ-producing CD4-lymphocytes characterise the early response to MTBVAC tuberculosis vaccine.细胞溶解性γδ T细胞和产生IFNγ的CD4淋巴细胞是对MTBVAC结核病疫苗早期反应的特征。
NPJ Vaccines. 2025 Mar 28;10(1):58. doi: 10.1038/s41541-025-01110-3.
5
Leucine zipper-based SAIM imaging identifies therapeutic agents to disrupt the cancer cell glycocalyx for enhanced immunotherapy.基于亮氨酸拉链的SAIM成像可识别破坏癌细胞糖萼以增强免疫治疗的治疗剂。
bioRxiv. 2024 Dec 6:2024.12.05.627089. doi: 10.1101/2024.12.05.627089.
6
Antibodies as key mediators of protection against .抗体作为预防 …… 的关键介质。
Front Immunol. 2024 Sep 2;15:1430955. doi: 10.3389/fimmu.2024.1430955. eCollection 2024.
7
Laboratory Features and Pathology of Cytokine Storm Syndromes.细胞因子风暴综合征的实验室特征和病理学。
Adv Exp Med Biol. 2024;1448:43-58. doi: 10.1007/978-3-031-59815-9_5.
8
BCG priming followed by a novel interleukin combination activates Natural Killer cells to selectively proliferate and become anti-tumour long-lived effectors.BCG 预刺激联合新型白细胞介素组合激活自然杀伤细胞，选择性增殖并成为抗肿瘤的长效效应细胞。
Sci Rep. 2024 Jun 7;14(1):13133. doi: 10.1038/s41598-024-62968-2.
9
Shedding Light on Intracellular Proteins using Flow Cytometry.流式细胞术解析细胞内蛋白。
Cell Biochem Biophys. 2024 Sep;82(3):1693-1707. doi: 10.1007/s12013-024-01338-1. Epub 2024 Jun 3.
10
Association of NK cell subsets and cytotoxicity with FCGR3A gene polymorphism in functional NK cell deficiency.NK 细胞亚群与细胞毒性与功能 NK 细胞缺陷中 FCGR3A 基因多态性的关联。
Rev Assoc Med Bras (1992). 2024 Feb 26;70(2):e20230872. doi: 10.1590/1806-9282.20230872. eCollection 2024.
本文引用的文献
1
Noncytotoxic functions of NK cells: direct pathogen restriction and assistance to adaptive immunity.自然杀伤细胞的非细胞毒性功能：直接限制病原体并辅助适应性免疫。
J Immunol. 2008 Jun 15;180(12):7785-91. doi: 10.4049/jimmunol.180.12.7785.
2
Line of attack: NK cell specificity and integration of signals.攻击路线：自然杀伤细胞的特异性与信号整合
Curr Opin Immunol. 2008 Jun;20(3):344-52. doi: 10.1016/j.coi.2008.03.005. Epub 2008 Apr 23.
3
Functions of natural killer cells.自然杀伤细胞的功能。
Nat Immunol. 2008 May;9(5):503-10. doi: 10.1038/ni1582.
4
Clinical analysis of human natural killer cells.人类自然杀伤细胞的临床分析
Methods Mol Biol. 2008;415:291-300. doi: 10.1007/978-1-59745-570-1_17.
5
Application of nine-color flow cytometry for detailed studies of the phenotypic complexity and functional heterogeneity of human lymphocyte subsets.九色流式细胞术在详细研究人淋巴细胞亚群的表型复杂性和功能异质性中的应用。
J Immunol Methods. 2008 Jan 31;330(1-2):64-74. doi: 10.1016/j.jim.2007.10.020. Epub 2007 Dec 4.
6
Spectrum, and clinical and functional implications of UNC13D mutations in familial haemophagocytic lymphohistiocytosis.家族性噬血细胞性淋巴组织细胞增生症中UNC13D突变的谱系、临床及功能意义
J Med Genet. 2008 Mar;45(3):134-41. doi: 10.1136/jmg.2007.054288. Epub 2007 Nov 9.
7
Defective cytotoxic lymphocyte degranulation in syntaxin-11 deficient familial hemophagocytic lymphohistiocytosis 4 (FHL4) patients.syntaxin-11缺陷型家族性噬血细胞性淋巴组织细胞增生症4型（FHL4）患者中细胞毒性淋巴细胞脱颗粒缺陷
Blood. 2007 Sep 15;110(6):1906-15. doi: 10.1182/blood-2007-02-074468. Epub 2007 May 24.
8
Prospects for the use of NK cells in immunotherapy of human cancer.自然杀伤细胞在人类癌症免疫治疗中的应用前景。
Nat Rev Immunol. 2007 May;7(5):329-39. doi: 10.1038/nri2073.
9
Intracellular cytokine optimization and standard operating procedure.细胞内细胞因子优化及标准操作程序。
Nat Protoc. 2006;1(3):1507-16. doi: 10.1038/nprot.2006.268.
10
Activation, coactivation, and costimulation of resting human natural killer cells.静息人自然杀伤细胞的激活、共激活和共刺激
Immunol Rev. 2006 Dec;214:73-91. doi: 10.1111/j.1600-065X.2006.00457.x.
Zotero 插件