Paneth Cells Are a Constitutive Source of IL-10 in Mouse Small Intestinal Organoids

The healthy gut masks a dynamic balance between pro- and anti-inflammatory activities, largely due to microbial factors in the lumen. IL-10 is vital among the anti-inflammatory mediators, yet confirming constitutive versus stimulated secretion in any cell type is difficult due to the cellular comple...

Full description

Saved in:
Bibliographic Details
Main Authors: Huong Nguyen, Francesca Di Cara, Jun Wang, Andrew W. Stadnyk
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Organoids
Subjects:
Online Access:https://www.mdpi.com/2674-1172/4/1/4
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850090978930589696
author Huong Nguyen
Francesca Di Cara
Jun Wang
Andrew W. Stadnyk
author_facet Huong Nguyen
Francesca Di Cara
Jun Wang
Andrew W. Stadnyk
author_sort Huong Nguyen
collection DOAJ
description The healthy gut masks a dynamic balance between pro- and anti-inflammatory activities, largely due to microbial factors in the lumen. IL-10 is vital among the anti-inflammatory mediators, yet confirming constitutive versus stimulated secretion in any cell type is difficult due to the cellular complexity in the gut. Seeking to determine whether intestinal epithelial cells are programmed to constitutively make IL-10, we confirmed that IL-10 mRNA was present in enteroids from C57BL/6 mice and IL-10 protein was co-localized with a Paneth cell marker but not with markers for goblet or tuft cells. Paneth cells positive for IL-10 also possessed apical and basal IL-10RA, while cells negative for IL-10 had only basal IL-10RA, suggesting a possible autocrine role for IL-10. Indeed, Paneth cells in IL-10 gene knockout (IL-10KO) enteroids possessed lower levels of anti-microbial protein mRNAs, which could not be restored by adding IL-10. Enteroids passaged onto Transwell<sup>®</sup> filters to form monolayers were treated with IL-10 and STAT3 phosphorylation was measured. Apically applied IL-10 resulted in a stronger STAT3 signal than basally applied cytokine. Our results indicate that a subpopulation of Paneth cells constitutively secrete IL-10 apically, which binds apical IL-10RA, impacting the expression of anti-microbial proteins unique to Paneth cells.
format Article
id doaj-art-04bbddc851c64549b0d4b198f232999d
institution DOAJ
issn 2674-1172
language English
publishDate 2025-02-01
publisher MDPI AG
record_format Article
series Organoids
spelling doaj-art-04bbddc851c64549b0d4b198f232999d2025-08-20T02:42:28ZengMDPI AGOrganoids2674-11722025-02-0141410.3390/organoids4010004Paneth Cells Are a Constitutive Source of IL-10 in Mouse Small Intestinal OrganoidsHuong Nguyen0Francesca Di Cara1Jun Wang2Andrew W. Stadnyk3Department of Microbiology & Immunology, Dalhousie University, Halifax, NS B3H 4R2, CanadaDepartment of Microbiology & Immunology, Dalhousie University, Halifax, NS B3H 4R2, CanadaDepartment of Microbiology & Immunology, Dalhousie University, Halifax, NS B3H 4R2, CanadaDepartment of Microbiology & Immunology, Dalhousie University, Halifax, NS B3H 4R2, CanadaThe healthy gut masks a dynamic balance between pro- and anti-inflammatory activities, largely due to microbial factors in the lumen. IL-10 is vital among the anti-inflammatory mediators, yet confirming constitutive versus stimulated secretion in any cell type is difficult due to the cellular complexity in the gut. Seeking to determine whether intestinal epithelial cells are programmed to constitutively make IL-10, we confirmed that IL-10 mRNA was present in enteroids from C57BL/6 mice and IL-10 protein was co-localized with a Paneth cell marker but not with markers for goblet or tuft cells. Paneth cells positive for IL-10 also possessed apical and basal IL-10RA, while cells negative for IL-10 had only basal IL-10RA, suggesting a possible autocrine role for IL-10. Indeed, Paneth cells in IL-10 gene knockout (IL-10KO) enteroids possessed lower levels of anti-microbial protein mRNAs, which could not be restored by adding IL-10. Enteroids passaged onto Transwell<sup>®</sup> filters to form monolayers were treated with IL-10 and STAT3 phosphorylation was measured. Apically applied IL-10 resulted in a stronger STAT3 signal than basally applied cytokine. Our results indicate that a subpopulation of Paneth cells constitutively secrete IL-10 apically, which binds apical IL-10RA, impacting the expression of anti-microbial proteins unique to Paneth cells.https://www.mdpi.com/2674-1172/4/1/4organoidPaneth cellinterleukin-10
spellingShingle Huong Nguyen
Francesca Di Cara
Jun Wang
Andrew W. Stadnyk
Paneth Cells Are a Constitutive Source of IL-10 in Mouse Small Intestinal Organoids
Organoids
organoid
Paneth cell
interleukin-10
title Paneth Cells Are a Constitutive Source of IL-10 in Mouse Small Intestinal Organoids
title_full Paneth Cells Are a Constitutive Source of IL-10 in Mouse Small Intestinal Organoids
title_fullStr Paneth Cells Are a Constitutive Source of IL-10 in Mouse Small Intestinal Organoids
title_full_unstemmed Paneth Cells Are a Constitutive Source of IL-10 in Mouse Small Intestinal Organoids
title_short Paneth Cells Are a Constitutive Source of IL-10 in Mouse Small Intestinal Organoids
title_sort paneth cells are a constitutive source of il 10 in mouse small intestinal organoids
topic organoid
Paneth cell
interleukin-10
url https://www.mdpi.com/2674-1172/4/1/4
work_keys_str_mv AT huongnguyen panethcellsareaconstitutivesourceofil10inmousesmallintestinalorganoids
AT francescadicara panethcellsareaconstitutivesourceofil10inmousesmallintestinalorganoids
AT junwang panethcellsareaconstitutivesourceofil10inmousesmallintestinalorganoids
AT andrewwstadnyk panethcellsareaconstitutivesourceofil10inmousesmallintestinalorganoids