{"title":"Enrichment of Neutrophils and Monocytes From the Liver Following Either Oral or Intravenous Listeria monocytogenes Infection","authors":"Michelle G. Pitts, Sarah E. F. D'Orazio","doi":"10.1002/cpim.102","DOIUrl":null,"url":null,"abstract":"<p><i>Listeria monocytogenes</i> is a foodborne pathogen that causes serious, often deadly, systemic disease in susceptible individuals such as neonates and the elderly. These facultative intracellular bacteria have been an invaluable tool in immunology research for more than three decades. Intravenous (i.v.) injection is the most commonly used transmission route in mice, but oral models of infection have also been developed in recent years, and these may be more appropriate for many studies. This article includes detailed instructions for use of either foodborne or i.v. inoculation of mice and discusses the rationale for choosing either model. Additionally, a protocol is provided for enrichment of neutrophils and monocytes from the infected liver in a manner that allows for determination of bacterial burden while still providing sufficient cells for use in flow cytometric analysis or in vitro assays. © 2020 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Foodborne <i>L. monocytogenes</i> infection</p><p><b>Support Protocol 1</b>: Preparing <i>L. monocytogenes</i> for foodborne infection</p><p><b>Basic Protocol 2</b>: Intravenous <i>L. monocytogenes</i> infection</p><p><b>Support Protocol 2</b>: Preparing <i>L. monocytogenes</i> for intravenous infection</p><p><b>Basic Protocol 3</b>: Enrichment of non-parenchymal cells from the infected liver</p>","PeriodicalId":10733,"journal":{"name":"Current Protocols in Immunology","volume":"130 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2020-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpim.102","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Immunology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpim.102","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Immunology and Microbiology","Score":null,"Total":0}
引用次数: 1
Abstract
Listeria monocytogenes is a foodborne pathogen that causes serious, often deadly, systemic disease in susceptible individuals such as neonates and the elderly. These facultative intracellular bacteria have been an invaluable tool in immunology research for more than three decades. Intravenous (i.v.) injection is the most commonly used transmission route in mice, but oral models of infection have also been developed in recent years, and these may be more appropriate for many studies. This article includes detailed instructions for use of either foodborne or i.v. inoculation of mice and discusses the rationale for choosing either model. Additionally, a protocol is provided for enrichment of neutrophils and monocytes from the infected liver in a manner that allows for determination of bacterial burden while still providing sufficient cells for use in flow cytometric analysis or in vitro assays. © 2020 Wiley Periodicals LLC.
Basic Protocol 1: Foodborne L. monocytogenes infection
Support Protocol 1: Preparing L. monocytogenes for foodborne infection
Basic Protocol 2: Intravenous L. monocytogenes infection
Support Protocol 2: Preparing L. monocytogenes for intravenous infection
Basic Protocol 3: Enrichment of non-parenchymal cells from the infected liver
口服或静脉注射单核增生李斯特菌感染后肝脏中性粒细胞和单核细胞的富集
单核细胞增生李斯特菌是一种食源性病原体,可在易感个体(如新生儿和老年人)中引起严重的、通常是致命的全身性疾病。三十多年来,这些兼性细胞内细菌一直是免疫学研究的宝贵工具。静脉注射是小鼠中最常用的传播途径,但近年来也开发了口服感染模型,这些模型可能更适合于许多研究。本文包括食源性或静脉接种小鼠的详细说明,并讨论了选择任何一种模型的基本原理。此外,提供了从感染肝脏富集中性粒细胞和单核细胞的方案,该方案允许确定细菌负荷,同时仍提供足够的细胞用于流式细胞术分析或体外测定。©2020 Wiley期刊有限责任公司基本方案1:食源性单核增生乳杆菌感染支持方案1:制备单核增生乳杆菌食源性感染支持方案2:制备单核增生乳杆菌静脉感染支持方案2:制备单核增生乳杆菌静脉感染基本方案3:从感染肝脏富集非实质细胞
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