Author:
Ichinose Mari,Suzuki Nobumi,Wang Tongtong,Wright Josephine A.,Lannagan Tamsin R. M.,Vrbanac Laura,Kobayashi Hiroki,Gieniec Krystyna,Ng Jia Q.,Ihara Souzaburo,Mavrangelos Chris,Hayakawa Yoku,Hughes Patrick,Worthley Daniel L.,Woods Susan L.
Abstract
AbstractThe intestinal stroma provides an important microenvironment for immune cell activation. The perturbation of this tightly regulated process can lead to excessive inflammation. We know that upregulated Toll-like receptor 4 (TLR4) in the intestinal epithelium plays a key role in the inflammatory condition of preterm infants, such as necrotizing enterocolitis (NEC). However, the surrounding stromal contribution to excessive inflammation in the pre-term setting awaits careful dissection. Ex vivo co-culture of embryonic day 14.5 (E14.5) or adult murine intestinal stromal cells with exogenous monocytes was undertaken. We also performed mRNAseq analysis of embryonic and adult stromal cells treated with vehicle control or lipopolysaccharide (LPS), followed by pathway and network analyses of differentially regulated transcripts. Cell characteristics were compared using flow cytometry and pHrodo red phagocytic stain, candidate gene analysis was performed via siRNA knockdown and gene expression measured by qPCR and ELISA. Embryonic stromal cells promote the differentiation of co-cultured monocytes to CD11bhighCD11chigh mononuclear phagocytes, that in turn express decreased levels of CD103. Global mRNAseq analysis of stromal cells following LPS stimulation identified TLR signaling components as the most differentially expressed transcripts in the immature compared to adult setting. We show that CD14 expressed by CD11b+CD45+ embryonic stromal cells is a key inducer of TLR mediated inflammatory cytokine production and phagocytic activity of monocyte derived cells. We utilise transcriptomic analyses and functional ex vivo modelling to improve our understanding of unique molecular cues provided by the immature intestinal stroma.
Funder
Japan Society for the Promotion of Science
Kanzawa Medical Research Foundation
Astellas Foundation for Research on Metabolic Disorders
Uehara Memorial Foundation
Takeda Science Foundation
Greaton International
National Health and Medical Research Council
Cancer Council SA Beat Cancer Project
Publisher
Springer Science and Business Media LLC
Cited by
1 articles.
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