{"title":"Protocol to study ex vivo T cell priming by conventional dendritic cells from the mouse spleen.","authors":"Mitra Ashayeripanah, Jose A Villadangos","doi":"10.1016/j.xpro.2024.103382","DOIUrl":null,"url":null,"abstract":"<p><p>Conventional dendritic cells (cDC) are professional antigen-presenting cells able to prime naive T cells. Here, we present a protocol for ex vivo T cell priming by murine splenic cDC. We describe the steps of injecting fluorescently labeled antigens to mice, purifying antigen-bearing cDC, and priming antigen-specific T cells ex vivo. This protocol is suitable for studying the T cell priming function of cDC in various murine models and helps factor in the effect of the microenvironment on cDC ability to uptake and process antigens. For complete details on the use and execution of this protocol, please refer to Ashayeripanah et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"5 4","pages":"103382"},"PeriodicalIF":1.3000,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11697547/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"STAR Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.xpro.2024.103382","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/12/11 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
Abstract
Conventional dendritic cells (cDC) are professional antigen-presenting cells able to prime naive T cells. Here, we present a protocol for ex vivo T cell priming by murine splenic cDC. We describe the steps of injecting fluorescently labeled antigens to mice, purifying antigen-bearing cDC, and priming antigen-specific T cells ex vivo. This protocol is suitable for studying the T cell priming function of cDC in various murine models and helps factor in the effect of the microenvironment on cDC ability to uptake and process antigens. For complete details on the use and execution of this protocol, please refer to Ashayeripanah et al.1.
传统的树突状细胞(cDC)是专业的抗原递呈细胞,能够为幼稚 T 细胞提供原代。在这里,我们介绍了一种由小鼠脾脏 cDC 进行体外 T 细胞诱导的方案。我们描述了向小鼠注射荧光标记抗原、纯化含抗原的 cDC 以及体内外启动抗原特异性 T 细胞的步骤。该方案适用于研究各种小鼠模型中 cDC 的 T 细胞启动功能,并有助于考虑微环境对 cDC 吸收和处理抗原能力的影响。有关使用和执行该方案的完整细节,请参阅 Ashayeripanah 等人的文章1。