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Transcriptomic analysis of human polarized macrophages: more than one role of alternative activation?

ABSTRACT: Background: Macrophages are a heterogeneous cell population which in response to the cytokine milieu polarize in either classically activated macrophages (M1) or alternatively activated macrophages (M2). This plasticity makes macrophages essential in regulating inflammation, immune response and tissue remodeling and a novel therapeutic target in inflammatory diseases such as atherosclerosis. The aim of the study was to describe the transcriptomic profiles of differently polarized human macrophages to generate new hypotheses on the biological function of the different macrophage subtypes. Methods and Results: M1 polarization was obtained by IFN-γ and LPS, M2a by IL-4, whereas IL-10 induced a “deactivated” state (M2c). Transcription profile of M1, M2a and M2c macrophages was performed at 6, 12 and 24h after polarization with Whole Human Genome Agilent Microarray technique. Gene Ontology (GO) classification revealed that M1 showed a significant up-regulation whereas M2a a down-regulation of GO terms involved in immunity and inflammation compared to resting macrophage (RM). Unexpectedly, canonical and non-canonical Wnt genes and gene groups, promoting inflammation and tissue remodeling, were up-regulated in M2a compared to RM. Key results were confirmed by real time-PCR. Conclusion: Results from gene expression profile confirmed the specific properties of differentially polarized macrophages. However, the enhanced expression of canonical and non-canonical Wnt pathways in M2a suggests a possible dual role for alternative activation in the modulation of low-grade inflammation.

ORGANISM(S): Homo sapiens

PROVIDER: GSE57614 | GEO | 2015/03/12

SECONDARY ACCESSION(S): PRJNA246958

REPOSITORIES: GEO

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Transcriptomic analysis of human polarized macrophages: more than one role of alternative activation?

Project description:Background: Macrophages are a heterogeneous cell population which in response to the cytokine milieu polarize in either classically activated macrophages (M1) or alternatively activated macrophages (M2). This plasticity makes macrophages essential in regulating inflammation, immune response and tissue remodeling and a novel therapeutic target in inflammatory diseases such as atherosclerosis. The aim of the study was to describe the transcriptomic profiles of differently polarized human macrophages to generate new hypotheses on the biological function of the different macrophage subtypes. Methods and Results: M1 polarization was obtained by IFN-γ and LPS, M2a by IL-4, whereas IL-10 induced a “deactivated” state (M2c). Transcription profile of M1, M2a and M2c macrophages was performed at 6, 12 and 24h after polarization with Whole Human Genome Agilent Microarray technique. Gene Ontology (GO) classification revealed that M1 showed a significant up-regulation whereas M2a a down-regulation of GO terms involved in immunity and inflammation compared to resting macrophage (RM). Unexpectedly, canonical and non-canonical Wnt genes and gene groups, promoting inflammation and tissue remodeling, were up-regulated in M2a compared to RM. Key results were confirmed by real time-PCR. Conclusion: Results from gene expression profile confirmed the specific properties of differentially polarized macrophages. However, the enhanced expression of canonical and non-canonical Wnt pathways in M2a suggests a possible dual role for alternative activation in the modulation of low-grade inflammation. Four-condition experiment, RM, M1, M2a, M2c. Three point of time course, three replicates for each condition. Dual color experiment, reference sample: human leucocytes

2015-03-12 | E-GEOD-57614 | biostudies-arrayexpress

In depth analysis reveals promotion of human macrophage polarization towards a proinflammatory M1 phenotype by TRAIL

Project description:The study aimed to investigate the impact of TRAIL on primary human M1, M2a and M2c macrophage polarization. Primary human monocyte-derived macrophages were pre-stimulated with TRAIL and then either left unpolarized (M0) or polarized into M1, M2a, or M2c phenotypes Then, RNA sequencing analysis was performed in 4 healthy donors including treated and control groups for one time point to analyze the differentially expressed M1 and M2 markers in TRAIL treated vs control groups.

2023-09-11 | GSE227737 | GEO

microRNA expression profiling of polarized macrophages

Project description:microRNAs are small non-coding RNAs that regulate gene expression at a post-translational level, and play a crucial role in the development of cells of the immune system. Macrophages are essential for generating inflammatory reactions upon tissue damage and encountering of invading pathogens, yet modulation of their immune responses is critical for maintaining tissue homeostasis. Macrophages can present different phenotypes, depending on the cytokine environment they encounter in the affected tissues. In this study, we have identified expression signatures of miRNAs that are differentially regulated during maturation of monocytes and polarization of macrophages by cytokines. We present a comprehensive characterization of miRNA expression in human monocytes and M1, M2a and M2c polarized macrophages, using next-generation Sequencing by Oligonucleotide Ligation and Detection (SOLiD). We have analyzed freshly isolated human monocytes and 5 day monocyte-derived macrophages unstimulated, or stimulated with IFNgamma+TNFalpha (M1), IL-4 (M2a) or IL-10 (M2c).

2014-01-14 | E-MTAB-1969 | biostudies-arrayexpress

RNA-sequencing analysis of db/db mice dorsal wounds treated with polarized macrophages conditioned media or control loaded bandages

Project description:We utilized bulk RNA-seq to profile the mRNA expression in day 10 wounds of db/db mice treated with control alginate bandages or bandages containing conditioned media from M0, M1, M2a and M2c polarized macrophages

2022-05-20 | GSE149419 | GEO

Genome-wide analysis of the gene expression of different subtype macrophages

Project description:Profiling the THP-1 derived macrophages, included M0, M1, M2a and M2c macrophages.

2021-07-05 | GSE116783 | GEO

Quantitative proteomic analysis of four subtypes of M2 macrophages

Project description:In this study, we used mass spectrometry and label-free quantification (LFQ) to characterize the global proteomics of polarized (M1, M2a, M2b, M2c, and M2d) and unpolarized (M0) phenotypes of macrophages from human THP-1 monocytes. The results described the biological functions of the four M2 macrophages subtypes and provided available references for identifying M2 macrophages or subtypes of M2 macrophages.

2022-08-12 | PXD022320 | Pride

CCR-5 using HIV-1 induces an anti-inflammatory profile in primary macrophages

Project description:Macrophages are important effector cells of the immune system and play an important role in mounting inflammatory responses. Macrophages can be activated by different stimuli in the tissue, either by cytokines produced by T helper cells (M1 or M2 polarization) or by the pathogens they encounter. Macrophages are also important target cells of HIV-1 and are preferentially infected by CCR5-using viruses. In this study, we investigated the ability of HIV-1 to induce changes in gene expression in unpolarized macrophages as well as in M1 or M2 polarized cells. We observed that CCR5-using HIV-1 regulates the expression of genes that are also regulated by IL-4 in macrophages. Genes regulated by HIV-1 infection and IL-4 polarization are involved in dampening pro-inflammatory responses in macrophages, which may facilitate HIV-1 to escape from detection by other immune cells. We also observed that changes in macrophage gene expression triggered by CCR5-using HIV-1 differed from those regulated by a CXCR4-using virus. This indicates that CCR5-using HIV-1 may be able to modulate macrophage gene expression to achieve successful replication. Our results provide insight in the complex interplay between HIV-1 and cells of the immune system. Polarized macrophages were obtained by stimulation of primary human monocytes with IFN-gamma (250 U/ml) in combination with TNF-alpha (12.5 ng/ml) (M1), IL-4 (50 ng/ml) (M2a), IL-10 (50 ng/ml) (M2c) for 5 days. Cells were inoculated for 24 hours with one of two HIV-1 strains (CCR5 or CXCR4 using HIV1) or their non replicating counterparts (heat inactivated virus). Macrophages that were not stimulated wiht cyokines or inoculates with HIV-1 were used as control. A total of 16 treatment conditions were tested in triplicate, for a total of 48 samples analysed.

2015-08-01 | E-GEOD-55029 | biostudies-arrayexpress

Gene expression analysis of myeloid cells cultured in the presence of fibronectin and/or anti-ILT3 16C5

Project description:RNA sequencing of monocyte-derived dendritic cells, tolerogenic dendritic cells, unpolarized macrophages, and M1, M2a, and M2c macrophages differentiated in vitro on PBS- or fibronectin-coated wells, in the presence of control IgG1 or anti-ILT3 16C5

2024-05-01 | GSE175354 | GEO

Gene Expression Analysis in Different Macrophage Phenotypes

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-Î³ (50 ng mLâ1), IFN-Î³ (50 ng mLâ1), IL-4 (10 ng mLâ1), IL-1Î² (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.

2016-08-10 | E-GEOD-85346 | biostudies-arrayexpress

Transcriptional profiling of human monocyte-derived macrophages

Project description:In this study we compared the response of human monocyte-derived macrophages differentiated with either granulocyte-macrophage colony-stimulating factor (GM-CSF) or macrophage colony-stimulating factor (M-CSF) to the most common activation stimuli: LPS plus interferon-γ to induce macrophage polarization towards the M1 type or IL-4 to induce macrophage polarization towards the M2a type. Additionally, IL-10 was used to drive M-CSF-primed macrophages into the M2c state. We used the the whole-human genome microarray to determine genes that were up- or downregulated by the activation stimuli in both macrophage lineages, with focus on genes implicated in immune response.

2018-05-22 | GSE61298 | GEO

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