RNF43 p.G659fs 通过 PI3K/AKT/mTOR 信号通路和 HLA-E 上调导致 MSI-high 结直肠癌中 NK 细胞功能障碍
RNF43 p.G659fs leads to natural killer cell dysfunction in MSI-high colorectal cancer through PI3K/AKT/mTOR signaling and HLA-E up-regulation.
CELL INTELLIGENCE · 肿瘤细胞治疗研究
肿瘤细胞治疗研究
一种共培养模型系统，用于利用工程化 NK 细胞系定量人乳腺癌细胞中的抗体依赖性细胞介导的细胞毒性
英文原题：A co-culture model system to quantify antibody-dependent cellular cytotoxicity in human breast cancer cells using an engineered natural killer cell line.
查看英文原题
A co-culture model system to quantify antibody-dependent cellular cytotoxicity in human breast cancer cells using an engineered natural killer cell line.
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中文摘要
目前的实验方案使用外周血单核细胞（PBMCs）在体外测量抗体依赖性细胞介导的细胞毒性（ADCC），但供者之间的分离和变异性限制了该方法的可行性和可重复性。在此，我们提出了一种标准化的共培养模型系统，用于量化人乳腺癌细胞上的ADCC。我们描述了构建稳定表达FCγRIIIa（CD16）的NK 细胞系的步骤，该受体是介导ADCC所必需的。随后，我们详细介绍了癌症-免疫共培养设置的步骤，以及随后的细胞毒性测量和分析。
展开英文摘要原文
Current protocols measure antibody-dependent cellular cytotoxicity (ADCC) in vitro using peripheral blood mononuclear cells (PBMCs), but isolation and variability among donors limit the viability and reproducibility of this approach.
Here, we present a standardized co-culture model system to quantify ADCC on human breast cancer cells.
We describe steps to engineer a natural killer cell line that stably expresses FCγRIIIa (CD16), required to mediate ADCC.
We then detail the steps for the cancer-immune co-culture setup, followed by cytotoxicity measurement and analysis.
论文信息
- 作者
- Vincken R、Ruiz-Saenz A
- 第一作者单位
- Department of Cell Biology, Erasmus University Medical Center Rotterdam, 3015 CN Rotterdam, the Netherlands. Electronic address: r.vincken@erasmusmc.nl.Netherlands
- 通讯作者单位
- Department of Cell Biology, Erasmus University Medical Center Rotterdam, 3015 CN Rotterdam, the Netherlands; Department of Medical Oncology, Erasmus University Medical Center Rotterdam, 3015 GD Rotterdam, the Netherlands. Electronic address: a.ruizsaenz@erasmusmc.nl.Netherlands
- 期刊
- STAR protocols2023 Jun 16
- 原文标识
- PubMed 37071532 · DOI 10.1016/j.xpro.2023.102224
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