猪肠道类器官作为哺乳动物正呼肠孤病毒3型感染体外模型的评价。

IF 1.5 3区 农林科学 Q2 VETERINARY SCIENCES
Se-A Lee, Hye Jeong Lee, Na-Yeon Gu, Yu-Ri Park, Eun-Ju Kim, Seok-Jin Kang, Bang-Hun Hyun, Dong-Kun Yang
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引用次数: 1

摘要

背景:哺乳动物正呼肠酶病毒3型(MRV3)在包括猪在内的许多哺乳动物物种中引起胃肠炎,已从患有严重腹泻的仔猪中分离出来。然而,使用猪源性细胞作为猪mrv3感染模型的研究很少。目的:本研究旨在建立猪肠道类器官(PIOs)并检测其对MRV3肠道感染的敏感性。方法:从小型猪空肠中分离并建立PIOs。建立的PIOs采用聚合酶链反应(PCR)和免疫荧光法(IFAs)进行表征,以确定小肠特异性基因和蛋白的表达,如Lgr5、LYZI、Mucin-2、ChgA和Villin。将暴露于根尖表面的单层pio和三维(3D) pio用MRV3感染2小时,用Dulbecco的磷酸盐缓冲盐水洗涤,观察。采用PCR和IFA检测证实病毒感染。我们采用定量实时逆转录- pcr来评估与肠上皮基因和抗病毒活性相关的病毒拷贝数和基因表达的变化。结果:所建立的PIOs具有肠道类器官的分子特征。感染的PIOs表现为增殖延迟和结构破坏。此外,MRV3感染改变了与肠上皮细胞相关的基因表达和抗病毒活性,这些影响在2D和3D模型中都观察到了。此外,两种模型的上清液中的病毒拷贝数都以时间依赖的方式增加。结论:我们认为PIOs可以作为一种体外模型来详细研究MRV3的感染机制,从而促进药物的开发。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Evaluation of porcine intestinal organoids as an <i>in vitro</i> model for mammalian orthoreovirus 3 infection.

Evaluation of porcine intestinal organoids as an <i>in vitro</i> model for mammalian orthoreovirus 3 infection.

Evaluation of porcine intestinal organoids as an <i>in vitro</i> model for mammalian orthoreovirus 3 infection.

Evaluation of porcine intestinal organoids as an in vitro model for mammalian orthoreovirus 3 infection.

Background: Mammalian orthoreovirus type 3 (MRV3), which is responsible for gastroenteritis in many mammalian species including pigs, has been isolated from piglets with severe diarrhea. However, the use of pig-derived cells as an infection model for swine-MRV3 has rarely been studied.

Objectives: This study aims to establish porcine intestinal organoids (PIOs) and examine their susceptibility as an in vitro model for intestinal MRV3 infection.

Methods: PIOs were isolated and established from the jejunum of a miniature pig. Established PIOs were characterized using polymerase chain reaction (PCR) and immunofluorescence assays (IFAs) to confirm the expression of small intestine-specific genes and proteins, such as Lgr5, LYZI, Mucin-2, ChgA, and Villin. The monolayered PIOs and three-dimensional (3D) PIOs, obtained through their distribution to expose the apical surface, were infected with MRV3 for 2 h, washed with Dulbecco's phosphate-buffered saline, and observed. Viral infection was confirmed using PCR and IFA. We performed quantitative real-time reverse transcription-PCR to assess changes in viral copy numbers and gene expressions linked to intestinal epithelial genes and antiviral activity.

Results: The established PIOs have molecular characteristics of intestinal organoids. Infected PIOs showed delayed proliferation with disruption of structures. In addition, infection with MRV3 altered the gene expression linked to intestinal epithelial cells and antiviral activity, and these effects were observed in both 2D and 3D models. Furthermore, viral copy numbers in the supernatant of both models increased in a time-dependent manner.

Conclusions: We suggest that PIOs can be an in vitro model to study the infection mechanism of MRV3 in detail, facilitating pharmaceutical development.

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来源期刊
Journal of Veterinary Science
Journal of Veterinary Science 农林科学-兽医学
CiteScore
3.10
自引率
5.60%
发文量
86
审稿时长
1.3 months
期刊介绍: The Journal of Veterinary Science (J Vet Sci) is devoted to the advancement and dissemination of scientific knowledge concerning veterinary sciences and related academic disciplines. It is an international journal indexed in the Thomson Scientific Web of Science, SCI-EXPANDED, Sci Search, BIOSIS Previews, Biological Abstracts, Focus on: Veterinary Science & Medicine, Zoological Record, PubMed /MEDLINE, Index Medicus, Pubmed Central, CAB Abstracts / Index Veterinarius, EBSCO, AGRIS and AGRICOLA. This journal published in English by the Korean Society of Veterinary Science (KSVS) being distributed worldwide.
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