Isolation, Characterization, and Culture of Intestinal Intraepithelial Lymphocytes

Olivia Jane James, Maud Vandereyken, Mahima Swamy (Lead / Corresponding author)

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

8 Citations (Scopus)
1027 Downloads (Pure)


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.

Original languageEnglish
Title of host publicationInnate Lymphoid Cells
Subtitle of host publicationMethods and Protocols
EditorsShoba Amarnath
Place of PublicationNew York
PublisherHumana Press
Number of pages12
ISBN (Electronic)9781071603383
ISBN (Print)9781071603376
Publication statusPublished - 2020

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • IEL
  • Innate-like lymphocytes
  • Intraepithelial lymphocytes
  • Small intestine
  • Unconventional T cells
  • γδ T cells

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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