A novel <i>in vitro</i> 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 <i>in vitro</i> model of the small intestinal epithelium in co-culture with 'gut-like' dendritic cells. Discovery immunology, 2 (1). kyad018-.

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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, <i>in vitro</i> models with sufficient complexity to understand these interactions have been lacking. Here, we outline the development of a co-culture model of <i>in vitro</i> 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-<i>trans</i>-retinoic acid (RA) resulted in the generation of a distinct population of conventional DCs expressing CD11b<sup>+</sup>SIRPα<sup>+</sup>CD103<sup>+/-</sup> (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:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3176443