title: Isolation and Flow Cytometry Analysis of Innate Lymphoid Cells from the Intestinal Lamina Propria.
abstract: The intestinal mucosa constitutes the biggest surface area of the body. It is constantly challenged by bacteria, commensal and pathogenic, protozoa, and food-derived irritants. In order to maintain homeostasis, a complex network of signaling circuits has evolved that includes contributions of immune cells. In recent years a subset of lymphocytes, which belong to the innate immune system, has caught particular attention. These so-called innate lymphoid cells (ILC) reside within the lamina propria of the small and large intestines and rapidly respond to environmental challenges. They provide immunity to various types of infections but may also contribute to organ homeostasis as they produce factors acting on epithelial cells thereby enhancing barrier integrity. Here, we describe how these cells can be isolated from their environment and provide an in-depth protocol how to visualize the various ILC subsets by flow cytometry.
authors.affiliation: Research Centre for Immunology and Institute of Medical Microbiology and Hygiene, University Medical Center of the Johannes Gutenberg-University Mainz, Obere Zahlbacher Strasse 67, 55131, Mainz, Germany.|Max Planck Institute for Immunobiology and Epigenetics, Stübeweg 51, 79108, Freiburg, Germany.
authors.forename: Konrad
authors.initials: K
authors.lastname: Gronke
authors.affiliation: Research Centre for Immunology and Institute of Medical Microbiology and Hygiene, University Medical Center of the Johannes Gutenberg-University Mainz, Obere Zahlbacher Strasse 67, 55131, Mainz, Germany.|Max Planck Institute for Immunobiology and Epigenetics, Stübeweg 51, 79108, Freiburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Kofoed-Nielsen
authors.affiliation: Research Centre for Immunology and Institute of Medical Microbiology and Hygiene, University Medical Center of the Johannes Gutenberg-University Mainz, Obere Zahlbacher Strasse 67, 55131, Mainz, Germany. andreas.diefenbach@charite.de.|Institute of Microbiology, Charité - University Medical Centre Berlin, Hindenburgdamm 27, 12203, Berlin, Germany. andreas.diefenbach@charite.de.
authors.forename: Andreas
authors.initials: A
authors.lastname: Diefenbach
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D015415:Biomarkers / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D017364:Collagenases / Q000737:chemistry; D003850:Deoxyribonuclease I / Q000737:chemistry; D010450:Endopeptidases / Q000737:chemistry; D005434:Flow Cytometry / Q000379:methods*; D015870:Gene Expression; D007113:Immunity, Innate; D018928:Immunity, Mucosal; D007413:Intestinal Mucosa / Q000166:cytology* / Q000276:immunology; D008214:Lymphocytes / Q000166:cytology* / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D000906:Antibodies; D015703:Antigens, CD; D015415:Biomarkers; D003850:Deoxyribonuclease I; D010450:Endopeptidases; D017364:Collagenases; C023139:dispase
keywords: ILC; Innate lymphoid cells; Lamina propria; Mucosa; Mucosal inflammation