用于共培养小鼠阴道上皮类器官和T细胞以诱导驻留记忆性CD8 T细胞分化的方案。

Protocol for the coculture of murine vaginal epithelial organoids and T cells to induce resident memory CD8 T cell differentiation.

作者信息

Ramprashad Julian C, Lin Ying, Beura Lalit K

机构信息

Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912, USA.

Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912, USA; Pathobiology Graduate Program, Brown University, Providence, RI 02912, USA.

出版信息

STAR Protoc. 2025 Aug 29;6(3):104054. doi: 10.1016/j.xpro.2025.104054.

DOI:10.1016/j.xpro.2025.104054

PMID:40884788

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12410536/

Abstract

Inducing robust resident memory T cell (TRM) establishment in mucosal tissues could enhance immunotherapy and vaccine efficacy. However, the factors influencing TRM formation are not fully understood. We present a protocol to coculture murine effector CD8 T cells with epithelial organoids, which serve as a reductionist model for investigating TRM differentiation in vitro. We describe steps for naive CD8 T cell activation, coculture of T cells with vaginal epithelial organoids (VEOs), and analysis of T cell phenotypes via flow cytometry. For complete details on the use and execution of this protocol, please refer to Ulibarri et al..

摘要

在黏膜组织中诱导强大的组织驻留记忆T细胞（TRM）形成可提高免疫治疗和疫苗的疗效。然而，影响TRM形成的因素尚未完全明确。我们提出了一种将小鼠效应CD8 T细胞与上皮类器官共培养的方案，上皮类器官可作为一种简化模型用于体外研究TRM分化。我们描述了初始CD8 T细胞激活、T细胞与阴道上皮类器官（VEO）共培养以及通过流式细胞术分析T细胞表型的步骤。有关本方案使用和实施的完整详细信息，请参考乌利巴里等人的研究。