title: Paneth cell extrusion and release of antimicrobial products is directly controlled by immune cell-derived IFN-γ.
abstract: Paneth cells (PCs) are terminally differentiated, highly specialized secretory cells located at the base of the crypts of Lieberkühn in the small intestine. Besides their antimicrobial function, PCs serve as a component of the intestinal stem cell niche. By secreting granules containing bactericidal proteins like defensins/cryptdins and lysozyme, PCs regulate the microbiome of the gut. Here we study the control of PC degranulation in primary epithelial organoids in culture. We show that PC degranulation does not directly occur upon stimulation with microbial antigens or bacteria. In contrast, the pro-inflammatory cytokine Interferon gamma (IFN-γ) induces rapid and complete loss of granules. Using live cell imaging, we show that degranulation is coupled to luminal extrusion and death of PCs. Transfer of supernatants from in vitro stimulated iNKT cells recapitulates degranulation in an IFN-γ-dependent manner. Furthermore, endogenous IFN-γ secretion induced by anti-CD3 antibody injection causes Paneth loss and release of goblet cell mucus. The identification of IFN-γ as a trigger for degranulation and extrusion of PCs establishes a novel effector mechanism by which immune responses may regulate epithelial status and the gut microbiome.
authors: Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical Centre Utrecht, 3584 CT Utrecht, Netherlands.
authors: Henner F
authors: HF
authors: Farin
authors: Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical Centre Utrecht, 3584 CT Utrecht, Netherlands.
authors: Wouter R
authors: WR
authors: Karthaus
authors: Antoni van Leeuwenhoek Hospital/Netherlands Cancer Institute, 1066 CX Amsterdam, Netherlands.
authors: Pekka
authors: P
authors: Kujala
authors: Section of Metabolic Diseases, Molecular Cancer Research, University Medical Center Utrecht, 3584 CG Utrecht, Netherlands.
authors: Maryam
authors: M
authors: Rakhshandehroo
authors: Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical Centre Utrecht, 3584 CT Utrecht, Netherlands.
authors: Gerald
authors: G
authors: Schwank
authors: Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical Centre Utrecht, 3584 CT Utrecht, Netherlands.
authors: Robert G J
authors: RG
authors: Vries
authors: Section of Metabolic Diseases, Molecular Cancer Research, University Medical Center Utrecht, 3584 CG Utrecht, Netherlands Netherlands Metabolomics Center, 2333 CC Leiden, Netherlands.
authors: Eric
authors: E
authors: Kalkhoven
authors: Department of Pediatric Gastroenterology, Wilhelmina Children's Hospital, University Medical Center Utrecht, 3584 EA Utrecht, Netherlands.
authors: Edward E S
authors: EE
authors: Nieuwenhuis
authors: Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical Centre Utrecht, 3584 CT Utrecht, Netherlands h.clevers@hubrecht.eu.
authors: Hans
authors: H
authors: Clevers
mesh_terms: D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D017252:CD3 Complex / Q000276:immunology; D015550:Cell Degranulation / Q000187:drug effects / Q000276:immunology*; D023082:Defensins / Q000276:immunology*; D020397:Goblet Cells / Q000166:cytology / Q000276:immunology; D007371:Interferon-gamma / Q000276:immunology*; D007421:Intestine, Small / Q000166:cytology / Q000276:immunology*; D051379:Mice; D018345:Mice, Knockout; D064307:Microbiota / Q000276:immunology; D009093:Mucus / Q000276:immunology; D009113:Muramidase / Q000276:immunology*; D055611:Natural Killer T-Cells / Q000276:immunology*; D009924:Organ Culture Techniques; D019879:Paneth Cells / Q000166:cytology / Q000276:immunology*
chemical_list: D000906:Antibodies; D017252:CD3 Complex; D023082:Defensins; D007371:Interferon-gamma; D009113:Muramidase