Delineating proinflammatory microenvironmental signals by ex vivo modeling of the immature intestinal stroma

Abstract The 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 inflammat...

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Autores principales:	Mari Ichinose, Nobumi Suzuki, Tongtong Wang, Josephine A. Wright, Tamsin R. M. Lannagan, Laura Vrbanac, Hiroki Kobayashi, Krystyna Gieniec, Jia Q. Ng, Souzaburo Ihara, Chris Mavrangelos, Yoku Hayakawa, Patrick Hughes, Daniel L. Worthley, Susan L. Woods
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/aebf352fe0324c3cb4059278bf6e460e
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spelling	oai:doaj.org-article:aebf352fe0324c3cb4059278bf6e460e2021-12-02T13:27:08ZDelineating proinflammatory microenvironmental signals by ex vivo modeling of the immature intestinal stroma10.1038/s41598-021-86675-42045-2322https://doaj.org/article/aebf352fe0324c3cb4059278bf6e460e2021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-86675-4https://doaj.org/toc/2045-2322Abstract The 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.Mari IchinoseNobumi SuzukiTongtong WangJosephine A. WrightTamsin R. M. LannaganLaura VrbanacHiroki KobayashiKrystyna GieniecJia Q. NgSouzaburo IharaChris MavrangelosYoku HayakawaPatrick HughesDaniel L. WorthleySusan L. WoodsNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Mari Ichinose Nobumi Suzuki Tongtong Wang Josephine A. Wright Tamsin R. M. Lannagan Laura Vrbanac Hiroki Kobayashi Krystyna Gieniec Jia Q. Ng Souzaburo Ihara Chris Mavrangelos Yoku Hayakawa Patrick Hughes Daniel L. Worthley Susan L. Woods Delineating proinflammatory microenvironmental signals by ex vivo modeling of the immature intestinal stroma
description	Abstract The 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.
format	article
author	Mari Ichinose Nobumi Suzuki Tongtong Wang Josephine A. Wright Tamsin R. M. Lannagan Laura Vrbanac Hiroki Kobayashi Krystyna Gieniec Jia Q. Ng Souzaburo Ihara Chris Mavrangelos Yoku Hayakawa Patrick Hughes Daniel L. Worthley Susan L. Woods
author_facet	Mari Ichinose Nobumi Suzuki Tongtong Wang Josephine A. Wright Tamsin R. M. Lannagan Laura Vrbanac Hiroki Kobayashi Krystyna Gieniec Jia Q. Ng Souzaburo Ihara Chris Mavrangelos Yoku Hayakawa Patrick Hughes Daniel L. Worthley Susan L. Woods
author_sort	Mari Ichinose
title	Delineating proinflammatory microenvironmental signals by ex vivo modeling of the immature intestinal stroma
title_short	Delineating proinflammatory microenvironmental signals by ex vivo modeling of the immature intestinal stroma
title_full	Delineating proinflammatory microenvironmental signals by ex vivo modeling of the immature intestinal stroma
title_fullStr	Delineating proinflammatory microenvironmental signals by ex vivo modeling of the immature intestinal stroma
title_full_unstemmed	Delineating proinflammatory microenvironmental signals by ex vivo modeling of the immature intestinal stroma
title_sort	delineating proinflammatory microenvironmental signals by ex vivo modeling of the immature intestinal stroma
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/aebf352fe0324c3cb4059278bf6e460e
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Delineating proinflammatory microenvironmental signals by ex vivo modeling of the immature intestinal stroma

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