Abstract
Intestinal intraepithelial lymphocytes (IEL) comprise distinct groups of innate-like and memory T cells that collectively form one of the largest T cell compartments in the body. IEL are located within the intestinal epithelium and are the first immune cells in the gut to interact with the food, microbiota, and pathogens that the gut is continually exposed to. IEL can respond rapidly to external insults to protect the small intestinal epithelium but are also considered regulatory cells that are important to maintain the homeostasis of the gut. However, the mechanisms of IEL activation and their interactions within the epithelium remain largely elusive. Indeed, IEL are not commonly evaluated even in studies of gut immunology, potentially because they are perceived as being difficult to isolate and study. In this protocol, we present a simplified method to isolate IEL from the murine small intestine and provide representative data for flow cytometric analyses of the different IEL subsets. We also outline two procedures for culturing IEL, which can permit functional studies and coculture with epithelial cells. These strategies should make studies of this large but enigmatic T cell compartment more accessible and open up understanding of homeostatic mechanisms in the intestine, and tissue-associated immunity.
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Acknowledgments
MV and MS are supported by the Wellcome Trust and Royal Society (Sir Henry Dale Fellowship to MS, 206246/Z/17/Z), and OJJ is supported by a Wellcome Trust PhD studentship (215309/Z/19/Z). We are grateful to the members of Prof. Adrian Hayday’s lab, King’s College London, from whom these protocols were modified, particularly Dr. Sarah Clarke who developed the protocol for long-term culture of IEL.
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James, O.J., Vandereyken, M., Swamy, M. (2020). Isolation, Characterization, and Culture of Intestinal Intraepithelial Lymphocytes. In: Amarnath, S. (eds) Innate Lymphoid Cells . Methods in Molecular Biology, vol 2121. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0338-3_13
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DOI: https://doi.org/10.1007/978-1-0716-0338-3_13
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0337-6
Online ISBN: 978-1-0716-0338-3
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