Extracellular CIRP Induces Macrophage Extracellular Trap Formation Via Gasdermin D Activation.
Extracellular CIRP Induces Macrophage Extracellular Trap Formation Via Gasdermin D Activation.
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DOI：
10.3389/fimmu.2021.780210
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发表时间：
2021
影响因子：
7.3
通讯作者：
Aziz M
中科院分区：
文献类型：
--
作者：
Lee Y;Reilly B;Tan C;Wang P;Aziz M
细胞外冷诱导RNA结合蛋白(ECIRP)是一种促进炎症和组织损伤的损伤相关分子模式。在细菌或病毒感染期间，巨噬细胞释放带有核蛋白和细胞质蛋白的DNA，称为巨噬细胞胞外陷阱(METS)。Gasdermin D(GSDMD)是一种成孔蛋白，参与了中性粒细胞胞外陷阱的形成。我们推测eCIRP通过激活GSDMD诱导MET的形成。用佛波酯(PMA)诱导人单核细胞系THP-1细胞分化，并用重组小鼠(RM)CIRP处理。用三种方法检测MET的形成：延时荧光显微镜(视频成像)、比色法和ELISA法。Western blotting检测裂解形式的GSDMD和caspase-1。SYTOX橙染色法显示，rmCIRP处理THP-1细胞后，MET的形成呈时间和剂量依赖性增加。胞外DNA、瓜氨酸组蛋白H3和髓过氧化物酶进一步证实了rmCIRP刺激形成的Met。与PBS处理的THP-1细胞相比，rmCIRP处理的THP-1细胞caspase-1和GSDMD的切割形式显著增加。分别用caspase-1、GSDMD抑制剂z-VAD-fmk和双硫仑处理巨噬细胞，可显著减少rmCIRP诱导的MET的形成。我们还利用原代细胞和小鼠腹膜巨噬细胞证实了rmCIRP诱导的MET的形成。这些数据清楚地表明，eCIRP是一种通过激活GSDMD和caspase-1而形成MET的新的诱导剂。
Extracellular cold-inducible RNA-binding protein (eCIRP) is a damage-associated molecular pattern promoting inflammation and tissue injury. During bacterial or viral infection, macrophages release DNA decorated with nuclear and cytoplasmic proteins known as macrophage extracellular traps (METs). Gasdermin D (GSDMD) is a pore-forming protein that has been involved in extracellular trap formation in neutrophils. We hypothesized that eCIRP induces MET formation by activating GSDMD. Human monocytic cell line THP-1 cells were differentiated with phorbol 12-myristate 13-acetate (PMA) and treated with recombinant murine (rm) CIRP. The MET formation was detected by three methods: time-lapse fluorescence microscopy (video imaging), colorimetry, and ELISA. Cleaved forms of GSDMD, and caspase-1 were detected by Western blotting. Treatment of THP-1 cells with rmCIRP increased MET formation as revealed by SYTOX Orange Staining assay in a time- and dose-dependent manner. METs formed by rmCIRP stimulation were further confirmed by extracellular DNA, citrullinated histone H3, and myeloperoxidase. Treatment of THP-1 cells with rmCIRP significantly increased the cleaved forms of caspase-1 and GSDMD compared to PBS-treated cells. Treatment of macrophages with caspase-1, and GSDMD inhibitors z-VAD-fmk, and disulfiram, separately, significantly decreased rmCIRP-induced MET formation. We also confirmed rmCIRP-induced MET formation using primary cells murine peritoneal macrophages. These data clearly show that eCIRP serves as a novel inducer of MET formation through the activation of GSDMD and caspase-1.
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影响因子：
64.8
作者：
Shi, Jianjin;Zhao, Yue;Shao, Feng
通讯作者：
Shao, Feng
影响因子：
3.5
作者：
Nishiyama, H;Higashitsuji, H;Fujita, J
通讯作者：
Fujita, J
影响因子：
82.9
作者：
通讯作者：
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DOI：
10.4049/jimmunol.2000091
发表时间：
2021-02-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Chen K;Murao A;Arif A;Takizawa S;Jin H;Jiang J;Aziz M;Wang P
通讯作者：
Wang P
影响因子：
3.1
作者：
Aulik, Nicole A.;Hellenbrand, Katrina M.;Czuprynski, Charles J.
通讯作者：
Czuprynski, Charles J.