Sus scrofa
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA188184 | ENA |
REPOSITORIES: ENA
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Project description:Human and mouse monocytes can be divided into 2 different sub-populations, using CD14-CD16 and Ly6C-CX3CR1 respectively. We investigated the pig monocytes sub-populations and found that all porcine monocyte express CD16 and CD172M-NM-1 but can be divided into 2 subpopulation using CD14 and the scavenger receptor CD163. The CD14hi-CD163low population resemble to the inflammatory monocytes whereas the CD14low-CD163hi display more a resident monocyte type. Pig monocyte can be differentiated into macrophages when cultured with rhCSF-1 and show an increase in size, granularity and autofluorescence, and express the common macrophage markers CD14, CD16 and CD172M-NM-1. Gene expression in these 2 sub-populations was profiled using the newly-developed and annotated pig whole genome snowball microarray, showing a distinct pattern between inflammatory and resident monocytes but this difference would be more a maturation process instead of two separate subsets. Furthermore, the expression of certain genes such as CD36, CLEC4E or TREM-1 proved to share the same pattern as human monocytes, quite different from mouse monocytes. These results emphasize the potential role of the pigs as a model for human inflammatory disease and will improved our knowledge on the mononuclear phagocyte system development. Porcine PBMCs were isolated from the blood of three seperate pigs, FACS sorted on expression of CD14 and CD163 and RNA isolated from each sample, a total of 6 microarrays were hybridised
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Project description:Monocyte maturation program into macrophages (MΦ) is well-defined in murine gut under homeostatic or inflammatory conditions. Obviously, in vivo tracking of monocytes in inflamed tissues remains difficult in humans. Furthermore, in vitro models fall short in generating the surrogates of transient extravasated tissue inflammatory monocytes. Here, we aimed to unravel environmental cues that replicate in vitro the human monocyte “waterfall” process by first generating tissue-like inflammatory monocytes, and then shifted them towards MΦ. Purified CD14+CD16- monocytes cultured with granulocyte-macrophage colony-stimulating factor (GM-CSF), IFNg and IL23 differentiated into CD14+CD163- cells displayed a monocyte-like morphology. The in vitro generated inflammatory CD14+CD163- cells (Infl mo-like), like the CD14+CD163- mo-like cells that accumulate in inflamed colon of Crohn’s disease patients, promoted IL-1β-dependent memory Th17 and Th17/Th1 responses. Next, in vitro generated Infl mo-like cells converted to functional CD163+ MΦ following exposure to TGFβ and IL10. Gene set enrichment analysis further revealed a shared molecular signature between converted CD163+ MΦ and MΦ detected in various inflamed non-lymphoid and lymphoid diseased tissues. Our findings propose a two-step in vitro culture that recapitulates human monocyte maturation cascade in inflamed tissue. Manipulating this process might open therapeutic avenues for chronic inflammatory disorders. Classical CD14+CD16- human monocytes were sorted and cultured for 6 days with GM-CSF+IFNg+IL23, leading to the generation of CD163- d6 cells. Following the 6 days culture with GM-CSF+IFNg+IL23, TGFβ+IL10 were added for another 6 days. This gave rise to the CD163+ d12 and CD163- d12 populations. We used microarray (Clariom D, Affymetrix) to observe molecular differences in the 3 in vitro generated populations: (1) CD163- d6 (2) CD163+ d12 (3) CD163- d12.
2020-06-23 | GSE149722 | GEO
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