Enhancing responsiveness of human jejunal enteroids to host and microbial stimuli.
cytokine
flagellin
histamine
human intestinal enteroids (HIEs)
toll-like receptors (TLR)
tumor necrosis factor (TNF)
Journal
The Journal of physiology
ISSN: 1469-7793
Titre abrégé: J Physiol
Pays: England
ID NLM: 0266262
Informations de publication
Date de publication:
08 2020
08 2020
Historique:
received:
09
12
2019
accepted:
11
05
2020
pubmed:
20
5
2020
medline:
16
2
2021
entrez:
20
5
2020
Statut:
ppublish
Résumé
Enteroids are a physiologically relevant model to examine the human intestine and its functions. Previously, the measurable cytokine response of human intestinal enteroids has been limited following exposure to host or microbial pro-inflammatory stimuli. Modifications to enteroid culture conditions facilitated robust human cytokine responses to pro-inflammatory stimuli. This new human enteroid culture methodology refines the ability to study microbiome:human intestinal epithelium interactions in the laboratory. The intestinal epithelium is the primary interface between the host, the gut microbiome and its external environment. Since the intestinal epithelium contributes to innate immunity as a first line of defence, understanding how the epithelium responds to microbial and host stimuli is an important consideration in promoting homeostasis. Human intestinal enteroids (HIEs) are primary epithelial cell cultures that can provide insights into the biology of the intestinal epithelium and innate immune responses. One potential limitation of using HIEs for innate immune studies is the relative lack of responsiveness to factors that stimulate epithelial cytokine production. We report technical refinements, including removal of extracellular antioxidants, to facilitate enhanced cytokine responses in HIEs. Using this new method, we demonstrate that HIEs have distinct cytokine profiles in response to pro-inflammatory stimuli derived from host and microbial sources. Overall, we found that host-derived cytokines tumour necrosis factor and interleukin-1α stimulated reactive oxygen species and a large repertoire of cytokines. In contrast, microbial lipopolysaccharide, lipoteichoic acid and flagellin stimulated a limited number of cytokines and histamine did not stimulate the release of any cytokines. Importantly, HIE-secreted cytokines were functionally active, as denoted by the ability of human blood-derived neutrophil to migrate towards HIE supernatant containing interleukin-8. These findings establish that the immune responsiveness of HIEs depends on medium composition and stimuli. By refining the experimental culture medium and creating an environment conducive to epithelial cytokine responses by human enteroids, HIEs can facilitate exploration of many experimental questions pertaining to the role of the intestinal epithelium in innate immunity.
Identifiants
pubmed: 32428244
doi: 10.1113/JP279423
pmc: PMC7674265
mid: NIHMS1603855
doi:
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
3085-3105Subventions
Organisme : NCI NIH HHS
ID : U01 CA170930
Pays : United States
Organisme : NIAID NIH HHS
ID : U01 AI124290
Pays : United States
Organisme : NIDDK NIH HHS
ID : P30 DK123704
Pays : United States
Organisme : NIAID NIH HHS
ID : F32 AI136404
Pays : United States
Organisme : NIH HHS
ID : T15LM007093
Pays : United States
Organisme : NIH HHS
ID : F32AI136404
Pays : United States
Organisme : NLM NIH HHS
ID : T15 LM007093
Pays : United States
Organisme : NIDDK NIH HHS
ID : R03 DK110270
Pays : United States
Organisme : NIH HHS
ID : 5T32DK007664-28
Pays : United States
Organisme : NIDDK NIH HHS
ID : T32 DK007664
Pays : United States
Organisme : NIH HHS
ID : P30-DK-56338
Pays : United States
Organisme : NIDDK NIH HHS
ID : P30 DK056338
Pays : United States
Organisme : NIH HHS
ID : F30DK112563
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK103759
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK115507
Pays : United States
Organisme : NIDDK NIH HHS
ID : F30 DK112563
Pays : United States
Organisme : NIH HHS
ID : U01CA170930
Pays : United States
Commentaires et corrections
Type : CommentIn
Informations de copyright
© 2020 The Authors. The Journal of Physiology © 2020 The Physiological Society.
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