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Toll样受体参与福氏耐格里阿米巴的识别。

Toll-like receptors participate in Naegleria fowleri recognition.

作者信息

Martínez-Castillo Moisés, Santos-Argumedo Leopoldo, Galván-Moroyoqui José Manuel, Serrano-Luna Jesús, Shibayama Mineko

机构信息

Department of Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies of the National Polytechnic Institute, Av. IPN 2508, 07360, Mexico City, Mexico.

Department of Molecular Biomedicine, Center for Research and Advanced Studies of the National Polytechnic Institute, Av. IPN 2508, 07360, Mexico City, Mexico.

出版信息

Parasitol Res. 2018 Jan;117(1):75-87. doi: 10.1007/s00436-017-5666-9. Epub 2017 Nov 11.

Abstract

Naegleria fowleri is a protozoan that invades the central nervous system and causes primary amoebic meningoencephalitis. It has been reported that N. fowleri induces an important inflammatory response during the infection. In the present study, we evaluated the roles of Toll-like receptors in the recognition of N. fowleri trophozoites by human mucoepithelial cells, analyzing the expression and production of innate immune response mediators. After amoebic interactions with NCI-H292 cells, the expression and production levels of IL-8, TNF-α, IL-1β, and human beta defensin-2 were evaluated by RT-PCR, ELISA, immunofluorescence, and dot blot assays, respectively. To determine whether the canonical signaling pathways were engaged, we used different inhibitors, namely, IMG-2005 for MyD88 and BAY 11-7085 for the nuclear factor NFkB. Our results showed that the expression and production of the pro-inflammatory cytokines and beta defensin-2 were induced by N. fowleri mainly through the canonical TLR4 pathway in a time-dependent manner.

摘要

福氏耐格里阿米巴是一种侵入中枢神经系统并引起原发性阿米巴脑膜脑炎的原生动物。据报道,福氏耐格里阿米巴在感染过程中会引发重要的炎症反应。在本研究中,我们评估了Toll样受体在人黏液上皮细胞识别福氏耐格里阿米巴滋养体中的作用,分析了固有免疫反应介质的表达和产生情况。在阿米巴与NCI-H292细胞相互作用后,分别通过逆转录聚合酶链反应(RT-PCR)、酶联免疫吸附测定(ELISA)、免疫荧光和斑点印迹分析评估白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和人β-防御素-2的表达和产生水平。为了确定经典信号通路是否被激活,我们使用了不同的抑制剂,即针对髓样分化因子88(MyD88)的IMG-2005和针对核因子κB(NFkB)的BAY 11-7085。我们的结果表明,福氏耐格里阿米巴主要通过经典的Toll样受体4(TLR4)通路以时间依赖性方式诱导促炎细胞因子和β-防御素-2的表达和产生。

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