CD14 and ALPK1 Affect Expression of Tight Junction Components and Proinflammatory Mediators upon Bacterial Stimulation in a Colonic 3D Organoid Model

CD14 and ALPK1 Affect Expression of Tight Junction Components and Proinflammatory Mediators upon Bacterial Stimulation in a Colonic 3D Organoid Model

Cd14 and Alpk1 both encode pathogen recognition receptors and are known candidate genes for affecting severity in inflammatory bowel diseases. CD14 acts as a coreceptor for bacterial lipopolysaccharide (LPS), while ALPK1 senses ADP-D-glycero-beta-D-manno-heptose, a metabolic intermediate of LPS bios...

Full description

Saved in:
Bibliographic Details
Published in:Stem cells international Vol. 2020; no. 2020; pp. 1 - 11
Main Authors: Buettner, Manuela, Liese, Andrea, Villarreal, Santiago Nahuel, Kalies, Stefan, Boyle, Erin C., zur Bruegge, Talke, Brooks, Pascal, Bleich, André
Format: Journal Article
Language:English
Published: Cairo, Egypt Hindawi Publishing Corporation 2020
Hindawi
John Wiley & Sons, Inc
Hindawi Limited
Subjects:
Adenosine diphosphate
Bacteria
Biosynthesis
CD14 antigen
Colon
Cytokines
E coli
Epithelial cells
Escherichia coli
Ethylenediaminetetraacetic acid
Experiments
Gastrointestinal diseases
Gene expression
Gene loci
Genes
Genotypes
Heptose
Immunohistochemistry
Inflammatory bowel disease
Inflammatory bowel diseases
Interleukin 1
Interleukin 10
Intestine
Kinases
Lipopolysaccharides
Mitogens
Organoids
Proteins
Receptors
Stem cells
Stimulation
Three dimensional models
Western blotting
Germany
California
United Kingdom > UK
United States > US
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Cd14 and Alpk1 both encode pathogen recognition receptors and are known candidate genes for affecting severity in inflammatory bowel diseases. CD14 acts as a coreceptor for bacterial lipopolysaccharide (LPS), while ALPK1 senses ADP-D-glycero-beta-D-manno-heptose, a metabolic intermediate of LPS biosynthesis. Intestinal barrier integrity can be influenced by CD14, whereas to date, the role of ALPK1 in maintaining barrier function remains unknown. We used colon-derived 3D organoids, first characterised for growth, proliferation, stem cell markers, and expression of tight junction (TJ) components using qPCR and immunohistochemistry. They showed characteristic crypt stem cells, apical shedding of dead cells, and TJ formation. Afterwards, organoids of different genotypes (WT, Il10-/-, Cd14-/-, and Alpk1-/-) were then stimulated with either LPS or Escherichia coli Nissle 1917 (EcN). Gene expression and protein levels of cytokines and TJ components were analysed. WT organoids increased expression of Tnfα and tight junction components. Cd14-/- organoids expressed significantly less Tnfα and Ocln after LPS stimulation than WT organoids but reacted similarly to WT organoids after EcN stimulation. In contrast, compared to WT, Alpk1-/- organoids showed decreased expression of different TJ and cytokine genes in response to EcN but not LPS. However, Western blotting revealed an effect of ALPK1 on TJ protein levels. These findings demonstrate that Cd14, but not Alpk1, alters the response to LPS stimulation in colonic epithelial cells, whereas Alpk1 is involved in the response upon bacterial challenge.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
Academic Editor: Sumanta Chatterjee
ISSN:1687-966X
1687-9678
1687-9678
DOI:10.1155/2020/4069354