Testing the Efficiency and Kinetics of Negative Selection Using Thymic Slices

Tyng An Zhou, Chia Lin Hsu, Ivan Lilyanov Dzhagalov*

*此作品的通信作者

研究成果: Chapter同行評審

3 引文 斯高帕斯(Scopus)

摘要

Central tolerance is an efficient barrier to autoimmunity and negative selection of self-reactive thymocytes is one of its major manifestations. Because of its importance, negative selection has been studied extensively through numerous in vitro and in vivo approaches that have tremendously increased our understanding of the process. Recently, in situ experimental systems using thymus slices have been developed that combine some of the advantages of in vitro assays such as ease of manipulation and high throughput with the existence of three dimensional mature thymus microenvironment. These approaches offer unprecedented opportunity to study negative selection. Here, we describe how thymic slices can be used to measure the kinetics and magnitude of negative selection. Taking the OT-1/Ova system as an example, we provide detailed guidance on cutting thymic slices, labeling and overlaying thymocytes on them and reading out the extent of negative selection by flow cytometry. The system can easily be adapted to evaluate the effects of various mutations or treatments on negative selection or to study the behavior of different cells in the thymus through time-lapse imaging.

原文English
主出版物標題Methods in Molecular Biology
發行者Humana Press Inc.
頁面205-219
頁數15
DOIs
出版狀態Published - 2020

出版系列

名字Methods in Molecular Biology
2111
ISSN(列印)1064-3745
ISSN(電子)1940-6029

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