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莫什科夫斯基内阿米巴作为展示和功能鉴定 Gal/GalNAc 凝集素中间亚基的潜在模式生物。

Entamoeba moshkovskii as a potential model organism for Gal/GalNAc lectin intermediate subunit exhibition and functional identification.

机构信息

Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Fudan University, Shanghai, China.

出版信息

Drug Discov Ther. 2024 Jul 9;18(3):178-187. doi: 10.5582/ddt.2024.01031. Epub 2024 May 22.

DOI:10.5582/ddt.2024.01031
PMID:38777764
Abstract

In humans, Entamoeba histolytica is the main pathogen causing various amoebiases, while E. moshkovskii falls between being a pathogen and non-pathogen. The two species have similar behavior patterns but differ significantly in pathogenicity, with previous studies and clinical data indicating that E. moshkovskii has a low level of pathogenicity. Meaningfully, the biological characteristics of E. moshkovskii make it a potential model organism and a protein display platform for studying the functions of important Entamoeba proteins. Here, an Amoeba-pcDNA3.1 vector capable of overexpressing E. histolytica-sourced Igl-C protein was constructed and successfully transfected into E. moshkovskii. High levels of expression of the Igl-C, EGFP, and NeoR genes were identified in Igl-C-transfected trophozoites using qRT-PCR, and they were subsequently confirmed using immunoblotting. Transfection of Igl-C protein improved the adherence and phagocytosis of E. moshkovskii, demonstrating that E. histolytica Igl mediated amoebic adhesion. Moreover, as a manifestation of protein virulence, the ability of post-transfected trophozoites to induce inflammation in host macrophages was also enhanced. In conclusion, this study utilizing the characteristics of E. moshkovskii confirmed its potential to serve as a model organism. E. moshkovskii could replace E. histolytica as the target of gene editing, allowing more efficient study of amoebic pathogenicity.

摘要

在人类中,溶组织内阿米巴是引起各种阿米巴病的主要病原体,而莫氏内阿米巴则处于病原体和非病原体之间。这两个物种具有相似的行为模式,但在致病性方面有很大的不同,以前的研究和临床数据表明,莫氏内阿米巴的致病性较低。有趣的是,莫氏内阿米巴的生物学特性使其成为研究重要内阿米巴蛋白功能的潜在模式生物和蛋白展示平台。在这里,构建了能够过表达溶组织内阿米巴源 Igl-C 蛋白的 Amoeba-pcDNA3.1 载体,并成功转染到莫氏内阿米巴中。使用 qRT-PCR 鉴定了 Igl-C 转染的滋养体中 Igl-C、EGFP 和 NeoR 基因的高表达水平,并通过免疫印迹进一步证实。Igl-C 蛋白的转染提高了莫氏内阿米巴的黏附和吞噬能力,表明溶组织内阿米巴 Igl 介导了阿米巴的黏附。此外,作为蛋白毒力的一种表现形式,转染后滋养体诱导宿主巨噬细胞炎症的能力也增强了。总之,本研究利用莫氏内阿米巴的特性证实了它作为模式生物的潜力。莫氏内阿米巴可以替代溶组织内阿米巴作为基因编辑的靶标,从而更有效地研究阿米巴的致病性。

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