A Simplified ELISpot Assay Protocol Used for Detection of Human Interleukin-4, Interleukin-13 and Interferon-γ Production in Response to the Contact Allergen Nickel

J. Minang, N. Ahlborg, M. Troye-Blomberg
{"title":"A Simplified ELISpot Assay Protocol Used for Detection of Human Interleukin-4, Interleukin-13 and Interferon-γ Production in Response to the Contact Allergen Nickel","authors":"J. Minang, N. Ahlborg, M. Troye-Blomberg","doi":"10.1159/000081568","DOIUrl":null,"url":null,"abstract":"Background: The ELISpot assay is a potential tool for in vitro diagnosis of contact allergy to nickel (Ni2+). A reduction of the assay time and work should further facilitate the development of ELISpot-based clinical and diagnostic applications. Objective: It was the aim of this study to evaluate a simplified ELISpot protocol utilizing plates precoated with capture monoclonal antibody (mAb) and one-step detection by an enzyme-labelled mAb. Methods: The frequency of Ni2+-induced IFN-γ-, IL-4- or IL-13-producing cells in human peripheral blood mononuclear cells from Ni2+-reactive and non-reactive subjects was determined. The simplified ELISpot was performed in parallel with a regular ELISpot assay utilizing overnight adsorption of capture mAb and detection in 2 steps. Results: Ni2+-induced IL-4 and IL-13 production was significantly greater in Ni2+-reactive subjects compared to the controls. The number of antigen-specific, cytokine-producing cells determined by the 2 different ELISpot protocols correlated well. Conclusion: The simplified ELISpot protocol provides a more rapid and easy alternative to the regular ELISpot, with a similar detection sensitivity for antigen-specific T-cell responses.","PeriodicalId":12086,"journal":{"name":"Exogenous Dermatology","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2004-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"10","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Exogenous Dermatology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1159/000081568","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 10

Abstract

Background: The ELISpot assay is a potential tool for in vitro diagnosis of contact allergy to nickel (Ni2+). A reduction of the assay time and work should further facilitate the development of ELISpot-based clinical and diagnostic applications. Objective: It was the aim of this study to evaluate a simplified ELISpot protocol utilizing plates precoated with capture monoclonal antibody (mAb) and one-step detection by an enzyme-labelled mAb. Methods: The frequency of Ni2+-induced IFN-γ-, IL-4- or IL-13-producing cells in human peripheral blood mononuclear cells from Ni2+-reactive and non-reactive subjects was determined. The simplified ELISpot was performed in parallel with a regular ELISpot assay utilizing overnight adsorption of capture mAb and detection in 2 steps. Results: Ni2+-induced IL-4 and IL-13 production was significantly greater in Ni2+-reactive subjects compared to the controls. The number of antigen-specific, cytokine-producing cells determined by the 2 different ELISpot protocols correlated well. Conclusion: The simplified ELISpot protocol provides a more rapid and easy alternative to the regular ELISpot, with a similar detection sensitivity for antigen-specific T-cell responses.
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
一种用于检测人对接触性变应原镍产生的白细胞介素-4、白细胞介素-13和干扰素-γ的简化ELISpot检测方案
背景:ELISpot法是体外诊断镍(Ni2+)接触性过敏的潜在工具。减少分析时间和工作将进一步促进基于elispot的临床和诊断应用的发展。目的:本研究的目的是评估一种简化的ELISpot方案,该方案利用预先包被捕获的单克隆抗体(mAb)的板和酶标记的mAb一步检测。方法:测定Ni2+反应性和非反应性人外周血单核细胞中产生IFN-γ-、IL-4-和il -13细胞的频率。简化ELISpot与常规ELISpot平行进行,利用隔夜吸附捕获mAb并分2步检测。结果:Ni2+诱导的IL-4和IL-13的产生在Ni2+反应的受试者中显著高于对照组。通过两种不同的ELISpot方案测定的抗原特异性细胞因子产生细胞的数量相关性良好。结论:与常规ELISpot相比,简化的ELISpot方案提供了一种更快速、更简便的替代方案,对抗原特异性t细胞反应的检测灵敏度相似。
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 去求助
来源期刊
自引率
0.00%
发文量
0
期刊最新文献
Acknowledgement to Referees for Exogenous Dermatology 2004 Contents Vol. 3, 2004 Author Index Vol. 3, 2004 Is There Evidence that Geraniol Causes Allergic Contact Dermatitis? Subject Index Vol. 3, 2004
×
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
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 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 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1