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研究小鼠脾脏常规树突状细胞体外T细胞启动的实验方案。

Protocol to study ex vivo T cell priming by conventional dendritic cells from the mouse spleen.

作者信息

Ashayeripanah Mitra, Villadangos Jose A

机构信息

Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, VIC 3000, Australia.

Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, VIC 3000, Australia; Department of Biochemistry and Pharmacology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Melbourne, VIC 3010, Australia.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103382. doi: 10.1016/j.xpro.2024.103382. Epub 2024 Dec 11.

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..

摘要

传统树突状细胞(cDC)是能够激活初始T细胞的专职抗原呈递细胞。在此,我们展示了一种通过小鼠脾脏cDC进行体外T细胞激活的方案。我们描述了向小鼠注射荧光标记抗原、纯化携带抗原的cDC以及在体外激活抗原特异性T细胞的步骤。该方案适用于研究各种小鼠模型中cDC的T细胞激活功能,并有助于考量微环境对cDC摄取和处理抗原能力的影响。有关此方案的使用和执行的完整详细信息,请参考阿沙耶里帕纳等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0d1f/11697547/25d68e6e1930/fx1.jpg

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