Cytokine Secretion and Pyroptosis of Thyroid Follicular Cells Mediated by Enhanced NLRP3, NLRP1, NLRC4, and AIM2 Inflammasomes Are Associated With Autoimmune Thyroiditis.
Cytokine Secretion and Pyroptosis of Thyroid Follicular Cells Mediated by Enhanced NLRP3, NLRP1, NLRC4, and AIM2 Inflammasomes Are Associated With Autoimmune Thyroiditis.
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NLRP3、NLRP1、NLRC4 和 AIM2 炎症小体增强介导的甲状腺滤泡细胞的细胞因子分泌和焦亡与自身免疫性甲状腺炎相关
DOI：
10.3389/fimmu.2018.01197
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发表时间：
2018
影响因子：
7.3
通讯作者：
Teng W
中科院分区：
文献类型：
--
作者：
Guo Q;Wu Y;Hou Y;Liu Y;Liu T;Zhang H;Fan C;Guan H;Li Y;Shan Z;Teng W
炎性小体介导白细胞介素-1 β（IL-β）和白细胞介素-18（IL-18）的成熟并导致细胞凋亡，其与各种自身免疫性疾病有关。本研究旨在探讨它们是否参与自身免疫性甲状腺炎（AIT）的发病机制。我们收集了50名AIT患者和50名性别和年龄匹配的对照组的甲状腺组织。血清游离T3、游离T4、促甲状腺激素、甲状腺过氧化物酶抗体（TPOAb）和甲状腺球蛋白抗体（TgAb）水平采用电化学发光免疫分析法测定。通过实时PCR和蛋白质印迹测定几种炎性体组分的表达，包括NOD样受体（NLR）家族pyrin结构域1（NLRP 1）、NLRP 3、CARD结构域4（NLRC 4），黑素瘤2（AIM 2）中缺失，包含半胱天冬酶募集结构域（ASC）的骨化相关斑点样蛋白，半胱天冬酶-1，IL-1β和IL-18。免疫组织化学法用于定位NLRP 1、NLRP 3、NLRC 4和AIM 2的表达。用肿瘤坏死因子-α（TNF-α）、干扰素-γ（IFN-γ）、白细胞介素-17 A、白细胞介素-6和聚（dA：dT）刺激Nthy-ori 3-1甲状腺细胞系。酶联免疫吸附法测定细胞培养上清中IL-18和IL-1β的含量，吸光光度法测定乳酸脱氢酶的含量。通过共聚焦免疫荧光显微镜分析来检查ASC斑点。流式细胞术检测细胞死亡情况，Western blot检测gasdermin D的N端结构域。AIT患者甲状腺组织中NLRP 1、NLRP 3、NLRC 4、AIM 2、ASC、caspase-1、pro IL-1β、pro IL-18、mRNA和蛋白的表达显著增加，并且还观察到caspase-1、IL-18和IL-1β的翻译后成熟增强。NLRP 1、NLRP 3、NLRC 4和AIM 2的表达主要位于邻近淋巴浸润区域的甲状腺滤泡细胞中。AIT组甲状腺NLRP 1和ASC mRNA水平与血清TPOAb和TgAb水平相关。TNF-α和IFN-γ对甲状腺细胞多种炎性体成分的表达具有启动作用。发现IFN-γ增强poly（dA：dT）诱导的细胞焦亡和生物活性IL-18的释放。我们的工作首次证明了多种炎性小体与AIT发病机制相关。NLRP 3、NLRP 1、NLRC 4、AIM 2炎性小体及其下游细胞因子可能是AIT潜在的治疗靶点和生物标志物。
Inflammasomes, which mediate maturation of interleukin-1β (IL-β) and interleukin-18 (IL-18) and lead to pyroptosis, have been linked to various autoimmune disorders. This study investigated whether they are involved in the pathogenesis of autoimmune thyroiditis (AIT). We collected thyroid tissues from 50 patients with AIT and 50 sex- and age-matched controls. Serum levels of free T3, free T4, thyrotropin, thyroid peroxidase antibody (TPOAb), and thyroglobulin antibody (TgAb) were measured by electrochemiluminescent immunoassays. Expression of several inflammasome components, the NOD-like receptor (NLR) family pyrin domain containing 1 (NLRP1), NLRP3, CARD-domain containing 4 (NLRC4), absent in melanoma 2 (AIM2), the apoptosis-associated speck-like protein that contains a caspase recruitment domain (ASC), caspase-1, IL-1β, and IL-18 was determined by real-time PCR and western blot. Immunohistochemistry was used to localize the expression of NLRP1, NLRP3, NLRC4, and AIM2. The Nthy-ori 3-1 thyroid cell line was stimulated with tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-17A, interleukin-6, and poly(dA:dT). The levels of IL-18 and IL-1β in the cell supernatant were measured by enzyme-linked immunosorbent assay, and lactate dehydrogenase was quantified by absorptiometry. ASC specks were examined by confocal immunofluorescence microscopic analysis. Cell death was examined by flow cytometry, and the N-terminal domain of gasdermin D was detected by western blot analysis. Expression of NLRP1, NLRP3, NLRC4, AIM2, ASC, caspase-1, pro IL-1β, pro IL-18, mRNA, and protein was significantly increased in thyroid tissues from patients with AIT, and enhanced posttranslational maturation of caspase-1, IL-18 and IL-1β was also observed. Expression of NLRP1, NLRP3, NLRC4, and AIM2 was localized mainly in thyroid follicular cells adjacent to areas of lymphatic infiltration. The thyroid mRNA level of NLRP1 and ASC was correlated to the serum TPOAb and TgAb levels in the AIT group. TNF-α and IFN-γ had a priming effect on the expression of multiple inflammasome components in thyroid cells. IFN-γ was found to strengthen poly(dA:dT)-induced cell pyroptosis and bioactive IL-18 release. Our work has demonstrated for the first time that multiple inflammasomes are associated with AIT pathogenesis. The identified NLRP3, NLRP1, NLRC4, AIM2 inflammasomes and their downstream cytokines may represent potential therapeutic targets and biomarkers of AIT.
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影响因子：
8.7
作者：
Man SM;Karki R;Kanneganti TD
通讯作者：
Kanneganti TD
影响因子：
4.9
作者：
Choulaki C;Papadaki G;Repa A;Kampouraki E;Kambas K;Ritis K;Bertsias G;Boumpas DT;Sidiropoulos P
通讯作者：
Sidiropoulos P
影响因子：
10.3
作者：
Marie, J.;Kovacs, D.;Cario-Andre, M.
通讯作者：
Cario-Andre, M.
影响因子：
17.1
作者：
Dombrowski Y;Peric M;Koglin S;Kammerbauer C;Göss C;Anz D;Simanski M;Gläser R;Harder J;Hornung V;Gallo RL;Ruzicka T;Besch R;Schauber J
通讯作者：
Schauber J
影响因子：
2.1
作者：
Mikos, Hanna;Mikos, Marcin;Niedziela, Marek
通讯作者：
Niedziela, Marek