A novel in vitro model of the small intestinal epithelium in co-culture with 'gut-like' dendritic cells.

Johnston, Luke J ORCID: 0000-0001-9656-8896, Barningham, Liam, Campbell, Eric L, Cerovic, Vuk, Duckworth, Carrie A ORCID: 0000-0001-7971-3561, Luu, Lisa ORCID: 0000-0002-7748-5961, Wastling, Jonathan, Derricott, Hayley and Coombes, Janine L (2023) A novel in vitro model of the small intestinal epithelium in co-culture with 'gut-like' dendritic cells. Discovery immunology, 2 (1). kyad018-.

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Official URL: http://dx.doi.org/10.1093/discim/kyad018

Abstract

Cross-talk between dendritic cells (DCs) and the intestinal epithelium is important in the decision to mount a protective immune response to a pathogen or to regulate potentially damaging responses to food antigens and the microbiota. Failures in this decision-making process contribute to the development of intestinal inflammation, making the molecular signals that pass between DCs and intestinal epithelial cells potential therapeutic targets. Until now, in vitro models with sufficient complexity to understand these interactions have been lacking. Here, we outline the development of a co-culture model of in vitro differentiated 'gut-like' DCs with small intestinal organoids (enteroids). Sequential exposure of murine bone marrow progenitors to Flt3L, granulocyte macrophage colony-stimulating factor (GM-CSF) and all-trans-retinoic acid (RA) resulted in the generation of a distinct population of conventional DCs expressing CD11b+SIRPα+CD103+/- (cDC2) exhibiting retinaldehyde dehydrogenase (RALDH) activity. These 'gut-like' DCs extended transepithelial dendrites across the intact epithelium of enteroids. 'Gut-like' DC in co-culture with enteroids can be utilized to define how epithelial cells and cDCs communicate in the intestine under a variety of different physiological conditions, including exposure to different nutrients, natural products, components of the microbiota, or pathogens. Surprisingly, we found that co-culture with enteroids resulted in a loss of RALDH activity in 'gut-like' DCs. Continued provision of GM-CSF and RA during co-culture was required to oppose putative negative signals from the enteroid epithelium. Our data contribute to a growing understanding of how intestinal cDCs assess environmental conditions to ensure appropriate activation of the immune response.

Item Type:	Article
Uncontrolled Keywords:	dendritic cell, enteroid, epithelial cell, flow cytometry, mucosal immunology
Divisions:	Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Infection, Veterinary and Ecological Sciences
Depositing User:	Symplectic Admin
Date Deposited:	25 Oct 2023 15:29
Last Modified:	13 Apr 2024 05:32
DOI:	10.1093/discim/kyad018
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3176443