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使用活细胞成像系统定量 NK 细胞对 3D 肿瘤球体的延时杀伤活性

英文原题:Quantification of timelapse 3D tumor spheroid killing activity of NK cells using a live-cell imaging system.

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Quantification of timelapse 3D tumor spheroid killing activity of NK cells using a live-cell imaging system.

PubMed 2025/10/14(内容时间) PLoS One Q2 · IF 2.8(JCR 2025)

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中文摘要

自然杀伤(NK)细胞是免疫系统的关键组成部分,负责识别和清除多种异常细胞,包括被病原体感染的细胞或转化为癌细胞的细胞。其强大的细胞毒性功能,包括直接释放细胞毒性颗粒、抗体依赖性细胞毒性(ADCC)以及表达凋亡诱导配体,使NK细胞成为癌症免疫治疗中一种有前景的治疗产品。

本研究提出了一套详细方案,利用活细胞成像系统评估NK细胞对三维(3D)肿瘤球体的细胞毒性,相较于传统2D培养,提供了更具生理相关性的模型。利用该3D球体模型,我们探索了NK细胞在不同效靶比(E:T)下对两种侵袭性实体癌细胞系——胆管癌(KKU-213A)和三阴性乳腺癌(MDA-MB-231)——杀伤活性的动态变化。

我们的研究结果揭示了NK细胞活性呈剂量依赖性增加,较高的E:T比产生更显著的肿瘤细胞死亡。实时成像进一步展示了两种细胞系在形态和细胞死亡模式上的明显差异,其中MDA-MB-231细胞对NK细胞细胞毒性表现出更快的反应。这些发现强调了3D球体模型和活细胞成像在研究NK细胞功能及推进基于NK细胞的免疫疗法开发中的实用性。本文详述的标准化方案为免疫监视和治疗应用提供了有价值的见解。

展开英文摘要原文

Natural killer (NK) cells are critical components of the immune system, responsible for recognizing and eliminating a wide range of abnormal cells, including those infected by pathogens or transformed into cancerous cells. Their potent cytotoxic functions, encompassing the direct release of cytotoxic granules, antibody-dependent cell cytotoxicity (ADCC), and the expression of apoptosis-inducing ligands, make NK cells a promising therapeutic product in cancer immunotherapy.

This study presents a detailed protocol for assessing NK cell-mediated cytotoxicity against three-dimensional (3D) tumor spheroids using a live-cell imaging system, offering a more physiologically relevant model compared to traditional 2D cultures. Utilizing this 3D spheroid model, we explored the dynamics of NK cell killing activity against two aggressive solid cancer cell lines, cholangiocarcinoma (KKU-213A) and triple-negative breast cancer (MDA-MB-231), across varying effector-to-target (E:T) ratios.

Our findings reveal a dose-dependent increase in NK cell activity, with higher E:T ratios yielding more pronounced tumor cell death. Real-time imaging further demonstrated distinct differences in the morphology and cell death patterns of the two cell lines, with MDA-MB-231 cells exhibiting a faster response to NK cell cytotoxicity.

These findings underscore the utility of 3D spheroid models and live-cell imaging for studying NK cell function and advancing the development of NK cell-based immunotherapies. The standardized protocol detailed herein provides a valuable insights into immune surveillance and therapeutic applications.

论文信息

作者
Srisantitham J、Thongsin N、Suwanpitak S、Wattanapanitch M
单位
Siriraj Center for Regenerative Medicine, Research Department, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.Thailand
期刊
PloS one2025
原文标识
PubMed 41086173 · DOI 10.1371/journal.pone.0334246