Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1061640 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR27446246_1.fastq.gz | Fastqsanger.gz | |||
| SRR27446246_2.fastq.gz | Fastqsanger.gz | |||
| SRR27446247_1.fastq.gz | Fastqsanger.gz | |||
| SRR27446247_2.fastq.gz | Fastqsanger.gz | |||
| SRR27442451_1.fastq.gz | Fastqsanger.gz |
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Project description:The relative contribution of polarized macrophages to the maintenance of tolerance is unknown. We examined their roles by in vivo adoptive transfer immunotherapy of M0, M1 and M2a macrophages as pre-treatment of colitis. In other experiments, M2a macrophages were used as pre-treatment or treatment of established colitis followed by immunotherapy with nTreg cells. Survival, weight gain, tissue infiltration, iTreg and Th17 cell development, T cell activation, and the frequency of proinflammatory cytokines were used as outcome measurements. Pre-treatment with M2a but not M1 macrophages increased the development of iTreg and Th17 cells. M2a macrophages used as pre-treatment or in treatment of established colitis allowed for successful therapy with nTreg cells. M1 and M2a macrophages have distinct gene expression profiles. M0, M1 and M2a macrophages were cell sorted and were used to generate total RNA for each array set, which was labeled and hybridized to Affymetrix Mouse Genome 430 2.0 GeneChips in accordance to the manufacturerâ??s protocol. Three sets of arrays were performed, and the results were averaged. The subset of probe sets whose expression increased or decreased by two-fold or more relative to M0 cells as a common standard was identified and used for further analysis.
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Project description:Macrophage activation is the main immunological process occurring during the development of several diseases, and the heterogeneity of macrophage activation or differentiation has been suggested to be involved in disease progression. In the present study, we attempted to identify molecules specifically expressed on human classically activated macrophages (M1) to investigate the significance of the M1-like phenotype in human diseases. Human monocyte-derived macrophages were differentiated into M1, M2a, M2b, and M2c phenotypes, and gene expression profiles were analyzed by cDNA microarray analysis and were used for bioinformatics examination. The gene expression profiles of murine macrophages were additionally evaluated. We identified guanylate-binding protein 5 (GBP5), which is associated with leucine-rich repeat protein 3-mediated inflammasome assembly in the M1 macrophages of both humans and mice. Notably, GBP5 protein expression was detected in cultured M1 macrophages by Western blot analysis. GBP5 is a useful candidate marker of the M1 phenotype. CD14+ monocytes from human PBMC were cultured with GM-CSF(10 ng mLâ1) or M-CSF (50 ng mLâ1) for seven days to differentiate into macrophages.To induce macrophage subtypes [M1, M1(-), M2a, M2b, and M2c], the macrophages were further stimulated for 24 h with LPS (10 ng mLâ1) + IFN-γ (50 ng mLâ1), IFN-γ (50 ng mLâ1), IL-4 (10 ng mLâ1), IL-1β (10 ng mLâ1), and IL-10 (10 ng mLâ1). Control macrophages (M0) were prepared by incubating for 24 h without additional factors.Two independent experiments were performed using different donors.
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