{"title":"Ascaris Mouse Model Protocols: Advancing Research on Larval Ascariasis Biology","authors":"Camila de Almeida Lopes, Jianbin Wang, Benjamin Liffner, Sabrina Absalon, Pedro H. Gazzinelli-Guimaraes","doi":"10.1002/cpz1.1074","DOIUrl":null,"url":null,"abstract":"<p>Ascariasis, caused by both <i>Ascaris lumbricoides</i> and <i>Ascaris suum</i>, is the most prevalent parasitic disease worldwide, affecting both human and porcine populations. However, due to the difficulties of assessing the early events of infection in humans, most studies of human ascariasis have been restricted to the chronic intestinal phase. Therefore, the <i>Ascaris</i> mouse model has become a fundamental tool for investigating the immunobiology and pathogenesis of the early infection stage referred to as larval ascariasis because of the model's practicality and ability to replicate the natural processes involved. The <i>Ascaris</i> mouse model has been widely used to explore factors such as infection resistance/susceptibility, liver inflammation, lung immune-mediated pathology, and co-infections and, notably, as a pivotal element in preclinical vaccine trials. Exploring the immunobiology of larval ascariasis may offer new insights into disease development and provide a substantial understanding of key components that trigger a protective immune response. This article focuses on creating a comprehensive guide for conducting <i>Ascaris</i> experimental infections in the laboratory as a foundation for future research efforts. © 2024 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Acquisition and embryonation of <i>Ascaris suum</i> eggs from adult females</p><p><b>Alternate Protocol</b>: Cleaning and purification of <i>Ascaris suum</i> from female <i>A. suum</i> uteri</p><p><b>Basic Protocol 2</b>: Preparation of <i>Ascaris suum</i> eggs and murine infection</p><p><b>Basic Protocol 3</b>: Measurement of larval burden and <i>Ascaris</i>-larva-induced pathogenesis</p><p><b>Basic Protocol 4</b>: <i>In vitro</i> hatching and purification of <i>Ascaris</i> L3 larvae</p><p><b>Support Protocol</b>: Preparation of crude antigen from <i>Ascaris</i> infectious stages</p><p><b>Basic Protocol 5</b>: Ultrastructure-expansion microscopy (U-ExM) of <i>Ascaris suum</i> larval stages</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"4 6","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2024-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpz1.1074","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
Ascariasis, caused by both Ascaris lumbricoides and Ascaris suum, is the most prevalent parasitic disease worldwide, affecting both human and porcine populations. However, due to the difficulties of assessing the early events of infection in humans, most studies of human ascariasis have been restricted to the chronic intestinal phase. Therefore, the Ascaris mouse model has become a fundamental tool for investigating the immunobiology and pathogenesis of the early infection stage referred to as larval ascariasis because of the model's practicality and ability to replicate the natural processes involved. The Ascaris mouse model has been widely used to explore factors such as infection resistance/susceptibility, liver inflammation, lung immune-mediated pathology, and co-infections and, notably, as a pivotal element in preclinical vaccine trials. Exploring the immunobiology of larval ascariasis may offer new insights into disease development and provide a substantial understanding of key components that trigger a protective immune response. This article focuses on creating a comprehensive guide for conducting Ascaris experimental infections in the laboratory as a foundation for future research efforts. © 2024 Wiley Periodicals LLC.
Basic Protocol 1: Acquisition and embryonation of Ascaris suum eggs from adult females
Alternate Protocol: Cleaning and purification of Ascaris suum from female A. suum uteri
Basic Protocol 2: Preparation of Ascaris suum eggs and murine infection
Basic Protocol 3: Measurement of larval burden and Ascaris-larva-induced pathogenesis
Basic Protocol 4: In vitro hatching and purification of Ascaris L3 larvae
Support Protocol: Preparation of crude antigen from Ascaris infectious stages
Basic Protocol 5: Ultrastructure-expansion microscopy (U-ExM) of Ascaris suum larval stages
蛔虫小鼠模型规程:推进幼虫蛔虫病生物学研究。
由蛔虫和猪蛔虫引起的蛔虫病是全球最流行的寄生虫病,同时影响人类和猪群。然而,由于难以评估人类感染的早期症状,大多数有关人类蛔虫病的研究仅限于慢性肠道阶段。因此,蛔虫小鼠模型因其实用性和复制自然过程的能力,已成为研究蛔虫幼虫感染早期免疫生物学和发病机制的基本工具。蛔虫小鼠模型已被广泛用于研究抗感染/易感性、肝脏炎症、肺部免疫介导的病理和合并感染等因素,尤其是作为临床前疫苗试验的关键因素。对蛔虫病幼虫免疫生物学的探索可能会为疾病的发展提供新的见解,并使人们对引发保护性免疫反应的关键成分有一个实质性的了解。本文重点介绍了在实验室进行蛔虫实验感染的综合指南,为今后的研究工作奠定了基础。© 2024 Wiley Periodicals LLC.基本方案 1:从成年雌性蛔虫卵中获取蛔虫卵并进行胚胎化 替代方案:基本方案 2:制备蛔虫卵和小鼠感染 基本方案 3:测量幼虫负担和蛔虫幼虫诱导的致病机制 基本方案 4:体外孵化和纯化蛔虫 L3 幼虫 支持方案:基本方案 5:蛔虫幼虫阶段的超微结构-扩展显微镜(U-ExM)。
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