{"title":"Protocol to evaluate virus-specific CD4<sup>+</sup> and CD8<sup>+</sup> T cell responses by spectral flow cytometry.","authors":"Xin-Jing Zhao, Sheng Zhang, Wang-Qian Wei, Qiang Xu, Chen-Long Lv, Guo-Lin Wang, Li-Qun Fang","doi":"10.1016/j.xpro.2024.103497","DOIUrl":null,"url":null,"abstract":"<p><p>T cell responses play an important role in viral clearance and prevention of infection. Here, we present a protocol to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific T cell responses using spectral flow cytometry. We describe steps for peripheral blood mononuclear cell (PBMC) thawing and stimulation with SARS-CoV-2 spike peptide pools. We then describe detailed procedures for cell confining, staining, and fixation. This protocol has potential application in spectral flow cytometry on other virus-specific T cell responses. For complete details on the use and execution of this protocol, please refer to Zhao et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"5 4","pages":"103497"},"PeriodicalIF":1.3000,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11699405/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"STAR Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.xpro.2024.103497","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/12/12 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
Abstract
T cell responses play an important role in viral clearance and prevention of infection. Here, we present a protocol to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific T cell responses using spectral flow cytometry. We describe steps for peripheral blood mononuclear cell (PBMC) thawing and stimulation with SARS-CoV-2 spike peptide pools. We then describe detailed procedures for cell confining, staining, and fixation. This protocol has potential application in spectral flow cytometry on other virus-specific T cell responses. For complete details on the use and execution of this protocol, please refer to Zhao et al.1.