An actin cytoskeletal barrier inhibits lytic granule release from natural killer cells in patients with Chediak-Higashi syndrome.
An actin cytoskeletal barrier inhibits lytic granule release from natural killer cells in patients with Chediak-Higashi syndrome.
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肌动蛋白细胞骨架屏障抑制了Chediak-Higashi综合征患者的天然杀伤细胞中裂解颗粒释放。
DOI：
10.1016/j.jaci.2017.10.040
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发表时间：
2018-09
期刊：
影响因子：
--
通讯作者：
Krzewski K
中科院分区：
文献类型：
--
作者：
Gil-Krzewska A;Saeed MB;Oszmiana A;Fischer ER;Lagrue K;Gahl WA;Introne WJ;Coligan JE;Davis DM;Krzewski K
Chediak-Higashi综合征(CHS)是一种罕见的疾病，由溶酶体运输调节基因(Lyst)的双等位基因突变引起，导致几种细胞类型中形成巨大的溶酶体或与溶酶体相关的细胞器。该病的特征是免疫缺陷和致命的噬血细胞淋巴组织细胞增多症，原因是细胞毒性淋巴细胞(包括自然杀伤细胞)功能受损。我们试图确定CHS患者NK细胞毒性受损的潜在生化原因。我们使用集群规则间隔短回文重复(CRISPR)技术生成了CHS的人类细胞模型。我们使用经典技术的组合来评估模型系统中的溶酶体功能和细胞活性，并使用超分辨率显微镜观察正常和Lyst缺陷的NK细胞中的F-肌动蛋白和裂解颗粒。人NK细胞系NK92mI中Lyst功能的丧失导致NK细胞杀伤作用的抑制，并复制了CHS细胞表型的其他方面，包括显著增大的裂解颗粒和有缺陷的胞吐功能，以及内溶酶体隔间的完整性受损。大颗粒的pH值为酸性，溶酶体酶活性正常，裂解颗粒胞吐所必需的蛋白质呈阳性。免疫突触肌动蛋白网孔的可视化显示，皮质肌动蛋白在细胞-细胞接触部位起到了分泌大颗粒的屏障作用。减少免疫突触处的皮质肌动蛋白密度或减小溶解颗粒的大小可以恢复Lyst缺陷的NK细胞脱颗粒和杀伤靶细胞的能力。皮质肌动蛋白和颗粒大小在NK细胞的细胞毒作用中起重要作用。我们提出的证据表明，突触下肌动蛋白的周期性是限制CHS患者NK细胞释放大溶解颗粒的重要因素，并可能成为药物干预的新靶点。
Chediak-Higashi syndrome (CHS) is a rare disorder caused by biallelic mutations in the lysosomal trafficking regulator gene (LYST), resulting in formation of giant lysosomes or lysosome-related organelles in several cell types. The disease is characterized by immunodeficiency and a fatal hemophagocytic lymphohistiocytosis caused by impaired function of cytotoxic lymphocytes, including natural killer (NK) cells. We sought to determine the underlying biochemical cause of the impaired cytotoxicity of NK cells in patients with CHS. We generated a human cell model of CHS using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technology. We used a combination of classical techniques to evaluate lysosomal function and cell activity in the model system and super-resolution microscopy to visualize F-actin and lytic granules in normal and LYST-deficient NK cells. Loss of LYST function in a human NK cell line, NK92mi, resulted in inhibition of NK cell cytotoxicity and reproduced other aspects of the CHS cellular phenotype, including the presence of significantly enlarged lytic granules with defective exocytosis and impaired integrity of endolysosomal compartments. The large granules had an acidic pH and normal activity of lysosomal enzymes and were positive for the proteins essential for lytic granule exocytosis. Visualization of the actin meshwork openings at the immunologic synapse revealed that the cortical actin acts as a barrier for secretion of such large granules at the cell-cell contact site. Decreasing the cortical actin density at the immunologic synapse or decreasing the lytic granule size restored the ability of LYST-deficient NK cells to degranulate and kill target cells. The cortical actin and granule size play significant roles in NK cell cytotoxic function. We present evidence that the periodicity of subsynaptic actin is an important factor limiting the release of large lytic granules from NK cells from patients with CHS and could be a novel target for pharmaceutical intervention.
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DOI：
10.1084/jem.20010938
发表时间：
2002-02-04
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Gerosa F;Baldani-Guerra B;Nisii C;Marchesini V;Carra G;Trinchieri G
通讯作者：
Trinchieri G
Rab27a及其效应子myrip将分泌颗粒与F-肌动蛋白联系起来，并控制其向释放位点的运动。
影响因子：
7.8
作者：
Desnos, Claire;Schonn, Jean-Sebastien;Huet, Sebastien;Tran, Viet Samuel;El-Amraoui, Aziz;Raposo, Graca;Fanget, Isabelle;Chapuis, Catherine;Menasche, Gael;de Saint Basile, Genevieve;Petit, Christine;Cribier, Sophie;Henry, Jean-Pierre;Darchen, Francois
通讯作者：
Darchen, Francois
影响因子：
20.3
作者：
Bryceson, Yenan T.;Pende, Daniela;Ehl, Stephan
通讯作者：
Ehl, Stephan
影响因子：
20.3
作者：
Elstak, Edo D.;Neeft, Maaike;van der Sluijs, Peter
通讯作者：
van der Sluijs, Peter
影响因子：
20.3
作者：
Fauriat, Cyril;Long, Eric O.;Bryceson, Yenan T.
通讯作者：
Bryceson, Yenan T.