Imaging burst kinetics and spatial coordination during serial killing by single natural killer cells
Imaging burst kinetics and spatial coordination during serial killing by single natural killer cells
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DOI：
10.1073/pnas.1221312110
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发表时间：
2013-04-16
影响因子：
11.1
通讯作者：
Mitchison, Timothy J.
中科院分区：
文献类型：
--
作者：
Choi, Paul J.;Mitchison, Timothy J.
细胞毒性淋巴细胞通过穿孔素-颗粒酶途径免疫识别和杀伤病毒感染细胞和癌细胞。传统的杀伤测定法测量固定时间的平均靶细胞溶解和高效应物：靶比率。这样的分析模糊了可能揭示新生理学的动力学细节。我们设计靶细胞以报告颗粒酶活性，使用非常低的效应子：靶比来观察潜在的连环杀伤，并进行低放大倍数延时成像以揭示单细胞水平上自然杀伤（NK）杀伤的时间依赖性统计数据。大多数杀伤发生在连续杀伤期间，单个NK细胞在6小时测定中杀死多达10个靶标。第一次杀伤比随后的杀伤慢，特别是在不良靶点上，或者当NK信号通路被部分抑制时。空间分析表明，连续杀人通常是相邻的。我们建议NK细胞整合来自先前和当前目标的信号，可能是通过同时接触。由此产生的爆发动力学和空间协调可以控制组织中NK细胞的活性。
Cytotoxic lymphocytes eliminate virus-infected and cancerous cells by immune recognition and killing through the perforin-granzyme pathway. Traditional killing assays measure average target cell lysis at fixed times and high effector: target ratios. Such assays obscure kinetic details that might reveal novel physiology. We engineered target cells to report on granzyme activity, used very low effector: target ratios to observe potential serial killing, and performed low magnification time-lapse imaging to reveal time-dependent statistics of natural killer (NK) killing at the single-cell level. Most kills occurred during serial killing, and a single NK cell killed up to 10 targets over a 6-h assay. The first kill was slower than subsequent kills, especially on poor targets, or when NK signaling pathways were partially inhibited. Spatial analysis showed that sequential kills were usually adjacent. We propose that NK cells integrate signals from the previous and current target, possibly by simultaneous contact. The resulting burst kinetics and spatial coordination may control the activity of NK cells in tissues.