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首页> 外文期刊>Journal of cell biology >Microclusters of inhibitory killer immunoglobulin–like receptor signaling at natural killer cell immunological synapses
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Microclusters of inhibitory killer immunoglobulin–like receptor signaling at natural killer cell immunological synapses

机译：天然杀伤细胞免疫突触中抑制性杀伤免疫球蛋白样受体信号转导的微簇

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摘要

We report the supramolecular organization of killer Ig–like receptor (KIR) phosphorylation using a technique applicable to imaging phosphorylation of any green fluorescent protein–tagged receptor at an intercellular contact or immune synapse. Specifically, we use fluorescence lifetime imaging (FLIM) to report F?rster resonance energy transfer (FRET) between GFP-tagged KIR2DL1 and a Cy3-tagged generic anti-phosphotyrosine monoclonal antibody. Visualization of KIR phosphorylation in natural killer (NK) cells contacting target cells expressing cognate major histocompatibility complex class I proteins revealed that inhibitory signaling is spatially restricted to the immune synapse. This explains how NK cells respond appropriately when simultaneously surveying susceptible and resistant target cells. More surprising, phosphorylated KIR was confined to microclusters within the aggregate of KIR, contrary to an expected homogeneous distribution of KIR signaling across the immune synapse. Also, yellow fluorescent protein–tagged Lck, a kinase important for KIR phosphorylation, accumulated in a multifocal distribution at inhibitory synapses. Spatial confinement of receptor phosphorylation within the immune synapse may be critical to how activating and inhibitory signals are integrated in NK cells.
机译：我们报告了一种杀伤性Ig样受体（KIR）磷酸化的超分子组织，该技术适用于细胞间接触或免疫突触中任何绿色荧光蛋白标记的受体的磷酸化成像。具体来说，我们使用荧光寿命成像（FLIM）报告GFP标签的KIR2DL1和Cy3标签的通用抗磷酸酪氨酸单克隆抗体之间的酯共振能转移（FRET）。在与表达同源主要组织相容性复合物I类蛋白的靶细胞接触的自然杀伤（NK）细胞中，KIR磷酸化的可视化揭示了抑制性信号在空间上局限于免疫突触。这解释了NK细胞在同时调查易感和抗性靶细胞时如何适当反应。更令人惊讶的是，磷酸化的KIR被限制在KIR聚集体内的微簇中，这与预期的KIR信号在整个免疫突触中的均匀分布相反。同样，带有黄色荧光蛋白标签的Lck（一种对KIR磷酸化很重要的激酶）在抑制性突触中以多焦点分布的形式积累。免疫突触中受体磷酸化的空间限制对于激活和抑制信号如何整合到NK细胞中可能至关重要。

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