In Vitro Evaluation of Toxicant Influences on the Immune System

Jane Kasten-Jolly, David A. Lawrence
{"title":"In Vitro Evaluation of Toxicant Influences on the Immune System","authors":"Jane Kasten-Jolly,&nbsp;David A. Lawrence","doi":"10.1002/cptx.95","DOIUrl":null,"url":null,"abstract":"<p>Culture of human peripheral blood mononuclear cells (PBMCs) still remains a convenient and sensitive method for measurement of a person's immune system health. Basic elements of the process, namely PBMC purification and culture medium formulation, were first reported in the late 1960s, and the utility of the method for clinical application was reported in the 1970s. Clinically, the approach can provide information about the ability of an individual's immune system to fight off attacks by various pathogens. Over the years, the method has undergone many improvements, which have been aided by advancements made in flow cytometry technology and the development of fluorescent reagents. The protocols presented here describe flow cytometry–based techniques for PBMC culture that can be employed to determine the impact of various environmental toxicants on the immune system. A major advantage of these procedures is that they will provide information about a toxicant or drug through <i>in vitro</i> methods. As the relationship between exposures to certain toxicants and an individual's response to vaccinations has been of concern, one of the protocols described shows how to test an environmental toxicant for potential modification of the immune response to vaccine antigen(s). © 2020 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Measurement of cell proliferation</p><p><b>Support Protocol 1</b>: Blood cell counting</p><p><b>Support Protocol 2</b>: Measurement of cell viability after culturing</p><p><b>Basic Protocol 2</b>: Identification of affected naïve/memory cell subsets</p>","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"84 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2020-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cptx.95","citationCount":"2","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols in toxicology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cptx.95","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 2

Abstract

Culture of human peripheral blood mononuclear cells (PBMCs) still remains a convenient and sensitive method for measurement of a person's immune system health. Basic elements of the process, namely PBMC purification and culture medium formulation, were first reported in the late 1960s, and the utility of the method for clinical application was reported in the 1970s. Clinically, the approach can provide information about the ability of an individual's immune system to fight off attacks by various pathogens. Over the years, the method has undergone many improvements, which have been aided by advancements made in flow cytometry technology and the development of fluorescent reagents. The protocols presented here describe flow cytometry–based techniques for PBMC culture that can be employed to determine the impact of various environmental toxicants on the immune system. A major advantage of these procedures is that they will provide information about a toxicant or drug through in vitro methods. As the relationship between exposures to certain toxicants and an individual's response to vaccinations has been of concern, one of the protocols described shows how to test an environmental toxicant for potential modification of the immune response to vaccine antigen(s). © 2020 Wiley Periodicals LLC.

Basic Protocol 1: Measurement of cell proliferation

Support Protocol 1: Blood cell counting

Support Protocol 2: Measurement of cell viability after culturing

Basic Protocol 2: Identification of affected naïve/memory cell subsets

毒物对免疫系统影响的体外评价
培养人外周血单个核细胞(PBMCs)仍然是一个方便和敏感的方法来测量一个人的免疫系统健康。该过程的基本要素,即PBMC纯化和培养基配方,于20世纪60年代末首次报道,并于20世纪70年代报道了该方法在临床应用中的实用性。在临床上,这种方法可以提供有关个体免疫系统抵御各种病原体攻击的能力的信息。多年来,该方法经历了许多改进,这得益于流式细胞术技术的进步和荧光试剂的发展。本文提出的方案描述了基于流式细胞术的PBMC培养技术,可用于确定各种环境毒物对免疫系统的影响。这些程序的一个主要优点是,它们将通过体外方法提供有关毒物或药物的信息。由于暴露于某些毒物与个人对接种疫苗的反应之间的关系一直受到关注,因此所描述的一项方案显示了如何测试环境毒物对疫苗抗原的免疫反应的潜在改变。©2020 Wiley期刊有限责任公司基本协议1:细胞增殖的测量支持协议1:血细胞计数支持协议2:培养后细胞活力的测量基本协议2:鉴定受影响naïve/记忆细胞亚群
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信