Monitoring the Interaction Between Dendritic Cells and T Cells In Vivo with LIPSTIC.
Monitoring the Interaction Between Dendritic Cells and T Cells In Vivo with LIPSTIC.
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DOI：
10.1007/978-1-0716-2938-3_5
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Victora GD
中科院分区：
文献类型：
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作者：
Pasqual G;Chudnovskiy A;Victora GD
不同细胞类型之间的相互作用是免疫功能的关键。传统上，相互作用已在体内研究活体双光子显微镜，但参与特定相互作用的细胞的分子表征是有限的，无法检索的细胞进行下游分析。我们最近开发了一种方法来标记在体内经历特定相互作用的细胞，我们称之为LIPSTIC（通过对细胞间接触进行分类来标记免疫伙伴关系）。在这里，我们提供了如何使用基因工程LIPSTIC小鼠跟踪树突状细胞（DC）和CD 4 + T细胞之间的CD 40-CD 40 L相互作用的详细说明。该方案需要动物实验和流式细胞术的专业知识。一旦实现小鼠杂交，需要3天或更长时间才能完成，这取决于研究人员希望研究的相互作用的动力学。
Interactions between different cell types are key for immune function. Traditionally, interactions have been investigated in vivo by intravital two-photon microscopy, but the molecular characterization of the cells participating in a specific interaction is limited by the inability to retrieve the cells for downstream analysis. We recently developed an approach to label cells undergoing specific interactions in vivo, which we called LIPSTIC (Labeling Immune Partnership by Sortagging Intercellular Contacts). Here, we provide detailed instructions on how to track CD40-CD40L interactions between dendritic cells (DCs) and CD4+ T cells using genetically engineered LIPSTIC mice. This protocol requires expertise in animal experimentation and multicolor flow cytometry. Once mouse crossing has been achieved, it takes 3 days or more to complete, depending on the kinetics of the interactions that the researcher wishes to investigate.