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使用质谱流式细胞术分析 NK 细胞与癌细胞的相互作用

英文原题:Profiling of Natural Killer Interactions With Cancer Cells Using Mass Cytometry.

查看英文原题

Profiling of Natural Killer Interactions With Cancer Cells Using Mass Cytometry.

PubMed 2023/05/09(内容时间) Lab Invest Q1 · IF 4.1(JCR 2025)

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中文摘要

我们开发了一种综合方法,利用质谱流式技术对与癌细胞共培养后外周血单个核细胞中免疫群体变化及杀伤活性进行功能评估。在本研究中,将包含43个标记的质谱流式 panel 应用于健康供者外周血单个核细胞来源的免疫细胞与多种癌细胞系的共培养体系。DNA含量结合经典CD45表面染色被用作共培养中免疫细胞(CD45高/DNA低)与血液系统癌细胞系(CD45低/DNA高)及实体癌细胞系(CD45阴性/DNA高)的设门参数。该策略无需特异性癌细胞标志物即可普遍区分癌细胞与免疫群体,并可同时评估两个群体的表型变化。质谱流式的使用可同时检测NK 细胞、自然杀伤T细胞和T细胞表型的变化,以及免疫群体在识别靶标后的脱颗粒、靶细胞中细胞毒性蛋白颗粒酶B含量的分析以及癌细胞死亡。这些发现不仅在NK-癌细胞相互作用的研究中,也在评估这些相互作用对其他免疫群体影响的研究和临床环境中,具有广泛的适用性,可用于表型分析和功能变化评估。

展开英文摘要原文

We developed a comprehensive method for functional assessment of the changes in immune populations and killing activity of peripheral blood mononuclear cells after cocultures with cancer cells using mass cytometry. In this study, a 43-marker mass cytometry panel was applied to a coculture of immune cells from healthy donors' peripheral blood mononuclear cells with diverse cancer cell lines. DNA content combined with classical CD45 surface staining was used as gating parameters for cocultures of immune cells (CD45 high /DNA low ) with hematological (CD45 low /DNA high ) and solid cancer cell lines (CD45 neg /DNA high ).

This strategy allows for universal discrimination of cancer cells from immune populations without the need for a specific cancer cell marker and simultaneous assessment of phenotypical changes in both populations. The use of mass cytometry allows for simultaneous detection of changes in natural killer, natural killer T cell, and T cell phenotypes and degranulation of immune populations upon target recognition, analysis of target cells for cytotoxic protein granzyme B content, and cancer cell death.

These findings have broad applicability in research and clinical settings with the aim to phenotype and assess functional changes following not only NK-cancer cell interactions but also the effect of those interactions on other immune populations.

论文信息

作者
Hallisey M、Dennis J、Gabriel EP、Masciarelli A、Chen J、Abrecht C、Brainard M、Marcotte WM
第一作者单位
Department of Medical Oncology, Center for Immuno-Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts.United States
通讯作者单位
Department of Medical Oncology, Center for Immuno-Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts. Electronic address: joanna_baginska@dfci.harvard.edu.United States
文献类型
非美国政府资助研究
期刊
Laboratory investigation; a journal of technical methods and pathology2023 Aug
原文标识
PubMed 37169083 · DOI 10.1016/j.labinv.2023.100174