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Testing the Efficiency and Kinetics of Negative Selection Using Thymic Slices

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Book cover T-Cell Receptor Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2111))

Abstract

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.

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Acknowledgments

We would like to thank Hsing-Kai Feng and Yun-Tzu Chen for help with taking the photos and carefully reviewing the manuscript. This work was supported by grants from MOST, 106-2320-B-010-026-MY3 (ILD), 107-2320-B-010-016-MY3 (ILD), 104-2628-B-010-002-MY4 (CLH), and 107-2320-B-010-020 (CLH), and a grant from the Yen Tjing Ling Medical Foundation, CI-108-5 (ILD).

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Authors and Affiliations

  1. Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan

    Tyng-An Zhou, Chia-Lin Hsu & Ivan Lilyanov Dzhagalov

Authors
  1. Tyng-An Zhou
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  2. Chia-Lin Hsu
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  3. Ivan Lilyanov Dzhagalov
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Corresponding author

Correspondence to Ivan Lilyanov Dzhagalov .

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Editors and Affiliations

  1. Department of Pathogen Biology, Huazhong University of Science and Technology, Wuhan, China

    Chaohong Liu

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Zhou, TA., Hsu, CL., Dzhagalov, I.L. (2020). Testing the Efficiency and Kinetics of Negative Selection Using Thymic Slices. In: Liu, C. (eds) T-Cell Receptor Signaling. Methods in Molecular Biology, vol 2111. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0266-9_17

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  • DOI: https://doi.org/10.1007/978-1-0716-0266-9_17

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0265-2

  • Online ISBN: 978-1-0716-0266-9

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