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

IF 1.9 Q3 PHARMACOLOGY & PHARMACY Drug Discoveries and Therapeutics Pub Date : 2024-07-09 Epub Date: 2024-05-22 DOI:10.5582/ddt.2024.01031
Qingshan Li, Meng Feng, Hongze Zhang, Hang Zhou, Xunjia Cheng
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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.

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将莫什科夫斯基恩塔米巴虫作为展示和鉴定 Gal/GalNAc 凝集素中间亚基功能的潜在模式生物。
在人类中,组织溶解恩塔米巴虫是导致各种阿米巴病的主要病原体,而莫什科夫斯基恩塔米巴虫则介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,但在致病性上有显著差异,以往的研究和临床数据表明,莫什科夫斯基伊氏菌的致病性较低。有意义的是,E. moshkovskii 的生物学特性使其成为研究重要恩塔米巴蛋白功能的潜在模式生物和蛋白质展示平台。在这里,我们构建了一种能够过表达来源于组织溶解虫的Igl-C蛋白的阿米巴-pcDNA3.1载体,并将其成功转染到莫什科夫斯基虫体内。利用 qRT-PCR 技术在转染 Igl-C 的滋养体中发现了 Igl-C、EGFP 和 NeoR 基因的高水平表达,随后利用免疫印迹技术对其进行了确认。转染Igl-C蛋白可提高E. moshkovskii的粘附性和吞噬能力,这表明组织溶解性伊丽莎白菌Igl介导了阿米巴粘附。此外,作为蛋白质毒力的一种表现,转染后滋养体诱导宿主巨噬细胞发炎的能力也得到了增强。总之,这项利用 E. moshkovskii 特性的研究证实了其作为模式生物的潜力。莫什科夫斯基伊蚊可以取代组织溶解伊蚊成为基因编辑的目标,从而更有效地研究阿米巴致病性。
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来源期刊
Drug Discoveries and Therapeutics
Drug Discoveries and Therapeutics PHARMACOLOGY & PHARMACY-
CiteScore
3.20
自引率
3.20%
发文量
51
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