A Novel Gastric Spheroid Co-culture Model Reveals Chemokine-Dependent Recruitment of Human Dendritic Cells to the Gastric Epithelium.
Cells, Cultured
Chemokines
/ genetics
Coculture Techniques
/ methods
Dendritic Cells
/ cytology
Epithelial Cells
/ cytology
Gastric Mucosa
/ cytology
Gene Expression Profiling
Gene Expression Regulation
Helicobacter Infections
/ genetics
Helicobacter pylori
/ immunology
Humans
Monocytes
/ cytology
Spheroids, Cellular
/ cytology
In Vitro Model
Mononuclear Phagocyte
Organoid
Stomach
Journal
Cellular and molecular gastroenterology and hepatology
ISSN: 2352-345X
Titre abrégé: Cell Mol Gastroenterol Hepatol
Pays: United States
ID NLM: 101648302
Informations de publication
Date de publication:
2019
2019
Historique:
received:
15
08
2017
revised:
13
02
2019
accepted:
20
02
2019
pubmed:
18
3
2019
medline:
9
4
2020
entrez:
18
3
2019
Statut:
ppublish
Résumé
Gastric dendritic cells (DCs) control the adaptive response to infection with Helicobacter pylori, a major risk factor for peptic ulcer disease and gastric cancer. We hypothesize that DC interactions with the gastric epithelium position gastric DCs for uptake of luminal H pylori and promote DC responses to epithelial-derived mediators. The aim of this study was to determine whether the gastric epithelium actively recruits DCs using a novel co-culture model of human gastric epithelial spheroids and monocyte-derived DCs. Spheroid cultures of primary gastric epithelial cells were infected with H pylori by microinjection. Co-cultures were established by adding human monocyte-derived DCs to the spheroid cultures and were analyzed for DC recruitment and antigen uptake by confocal microscopy. Protein array, gene expression polymerase chain reaction array, and chemotaxis assays were used to identify epithelial-derived chemotactic factors that attract DCs. Data from the co-culture model were confirmed using human gastric tissue samples. Human monocyte-derived DCs co-cultured with gastric spheroids spontaneously migrated to the gastric epithelium, established tight interactions with the epithelial cells, and phagocytosed luminally applied H pylori. DC recruitment was increased upon H pylori infection of the spheroids and involved the activity of multiple chemokines including CXCL1, CXCL16, CXCL17, and CCL20. Enhanced chemokine expression and DC recruitment to the gastric epithelium also was observed in H pylori-infected human gastric tissue samples. Our results indicate that the gastric epithelium actively recruits DCs for immunosurveillance and pathogen sampling through chemokine-dependent mechanisms, with increased recruitment upon active H pylori infection.
Sections du résumé
BACKGROUND & AIMS
Gastric dendritic cells (DCs) control the adaptive response to infection with Helicobacter pylori, a major risk factor for peptic ulcer disease and gastric cancer. We hypothesize that DC interactions with the gastric epithelium position gastric DCs for uptake of luminal H pylori and promote DC responses to epithelial-derived mediators. The aim of this study was to determine whether the gastric epithelium actively recruits DCs using a novel co-culture model of human gastric epithelial spheroids and monocyte-derived DCs.
METHODS
Spheroid cultures of primary gastric epithelial cells were infected with H pylori by microinjection. Co-cultures were established by adding human monocyte-derived DCs to the spheroid cultures and were analyzed for DC recruitment and antigen uptake by confocal microscopy. Protein array, gene expression polymerase chain reaction array, and chemotaxis assays were used to identify epithelial-derived chemotactic factors that attract DCs. Data from the co-culture model were confirmed using human gastric tissue samples.
RESULTS
Human monocyte-derived DCs co-cultured with gastric spheroids spontaneously migrated to the gastric epithelium, established tight interactions with the epithelial cells, and phagocytosed luminally applied H pylori. DC recruitment was increased upon H pylori infection of the spheroids and involved the activity of multiple chemokines including CXCL1, CXCL16, CXCL17, and CCL20. Enhanced chemokine expression and DC recruitment to the gastric epithelium also was observed in H pylori-infected human gastric tissue samples.
CONCLUSIONS
Our results indicate that the gastric epithelium actively recruits DCs for immunosurveillance and pathogen sampling through chemokine-dependent mechanisms, with increased recruitment upon active H pylori infection.
Identifiants
pubmed: 30878664
pii: S2352-345X(19)30026-8
doi: 10.1016/j.jcmgh.2019.02.010
pmc: PMC6599165
pii:
doi:
Substances chimiques
Chemokines
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
157-171.e3Subventions
Organisme : NIDDK NIH HHS
ID : K01 DK097144
Pays : United States
Organisme : NIGMS NIH HHS
ID : P30 GM110732
Pays : United States
Organisme : NIDDK NIH HHS
ID : R03 DK107960
Pays : United States
Organisme : NIH HHS
ID : R44 OD012083
Pays : United States
Commentaires et corrections
Type : CommentIn
Informations de copyright
Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.
Références
Mucosal Immunol. 2015 May;8(3):533-44
pubmed: 25249167
EMBO Mol Med. 2013 May;5(5):776-94
pubmed: 23606583
Blood. 2012 Nov 1;120(18):3741-9
pubmed: 22972984
Elife. 2017 Nov 07;6:
pubmed: 29110754
Science. 2005 Jan 14;307(5707):254-8
pubmed: 15653504
Mucosal Immunol. 2015 Jul;8(4):799-814
pubmed: 25492476
Nat Immunol. 2001 Apr;2(4):361-7
pubmed: 11276208
J Gastroenterol. 2016 Mar;51(3):206-13
pubmed: 26800996
Mol Pharm. 2010 Dec 6;7(6):2103-19
pubmed: 20809575
Helicobacter. 2012 Sep;17 Suppl 1:9-15
pubmed: 22958149
Blood. 2009 Apr 9;113(15):3418-27
pubmed: 19176316
Pathog Dis. 2013 Feb;67(1):46-53
pubmed: 23620119
Cell Rep. 2017 Dec 26;21(13):3860-3872
pubmed: 29281833
Innate Immun. 2017 Feb;23(2):165-174
pubmed: 27913793
Immunology. 2017 Dec;152(4):613-627
pubmed: 28746740
Gastroenterology. 2009 May;136(6):1863-73
pubmed: 19457415
Helicobacter. 2009 Jun;14(3):208-22
pubmed: 19702851
J Leukoc Biol. 2017 May;101(5):1169-1180
pubmed: 28087652
J Immunol. 2013 Jun 15;190(12):6626-34
pubmed: 23686492
Eur J Immunol. 2002 Jan;32(1):231-42
pubmed: 11782014
Sci Rep. 2017 Mar 27;7:45270
pubmed: 28345602
Cell Tissue Res. 2018 Feb;371(2):293-307
pubmed: 29178040
J Leukoc Biol. 2010 Oct;88(4):747-56
pubmed: 20651299
Gastroenterology. 2010 Mar;138(3):1046-54
pubmed: 19931266
J Exp Med. 2006 Dec 25;203(13):2841-52
pubmed: 17145958
Gastroenterology. 2011 Sep;141(3):929-38
pubmed: 21699795
Gut Microbes. 2010 Mar;1(2):109-113
pubmed: 21326919
Mucosal Immunol. 2010 May;3(3):260-9
pubmed: 20237463
EMBO Rep. 2009 Nov;10(11):1242-9
pubmed: 19820695
Immunology. 1991 Feb;72(2):239-45
pubmed: 2016121
Infect Immun. 2014 Jul;82(7):2881-9
pubmed: 24778119
Infect Immun. 2006 Aug;74(8):4624-33
pubmed: 16861650
Eur J Immunol. 2006 Apr;36(4):864-74
pubmed: 16544275
Immunology. 2008 Feb;123(2):197-208
pubmed: 17655740
Mucosal Immunol. 2009 Jul;2(4):340-50
pubmed: 19387433
Infect Immun. 2001 Jan;69(1):81-8
pubmed: 11119492
Proc Natl Acad Sci U S A. 2017 Jan 31;114(5):E751-E760
pubmed: 28096401
Gut. 2016 Feb;65(2):202-13
pubmed: 25539675
J Immunol. 2011 Jun 15;186(12):7067-79
pubmed: 21551359
BMC Med Genomics. 2013 Oct 11;6:41
pubmed: 24119614
Infect Immun. 1994 Jun;62(6):2609-13
pubmed: 8188385
Oncoimmunology. 2016 Apr 22;5(6):e1160979
pubmed: 27471636
J Exp Med. 1998 Jul 20;188(2):373-86
pubmed: 9670049
Gut. 2009 Nov;58(11):1481-9
pubmed: 19570762
Gastroenterology. 2015 Jan;148(1):126-136.e6
pubmed: 25307862
Blood. 2012 Jul 19;120(3):572-80
pubmed: 22589473
Nat Methods. 2012 Jul;9(7):671-5
pubmed: 22930834
J Immunol. 2018 Jul 15;201(2):714-724
pubmed: 29875152
BMC Microbiol. 2015 Apr 1;15(1):79
pubmed: 25887178