Homo sapiens
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA121199 | ENA |
REPOSITORIES: ENA
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Project description:Chaperonin 60 (Cpn60) is a prototypic molecular chaperone essential for cellular function due to its protein folding actions. However, over the past decade it has been established that Cpn60 can be released by human cells and by certain bacteria to act as an extracellular signalling protein. Mycobacterium tuberculosis produces two Cpn60 proteins: Cpn60.1 and Cpn60.2. We recently generated a M. tuberculosis mutant with an inactivated cpn60.1 gene and demonstrated that granuloma formation was impaired after murine/guinea pig infection. This finding suggested that Cpn60.1 may interact with the cellular organisation of the host response to M. tuberculosis bacilli. In this study, we report that recombinant M. tuberculosis Cpn60.1 has both pro- and anti-inflammatory effects on human circulating monocytes. At high concentrations, recombinant Cpn60.1 induces the synthesis of TNF-α, IL6, and IL8, and promotes the phosphorylation of NF-κBp65, p44/42MAPK and p38 MAPK. At lower concentrations M. tuberculosis Cpn60.1 inhibits lipopolysaccharide-induced release of TNF-α, and monocyte transcriptional activation program. Both effects are abrogated by proteolysis of Cpn60.1 and therefore cannot be attributed to contamination with lipopolysaccharide. Competition with LPS for binding to a common receptor, the release of IL-10 or down-regulation of TLR4 on the cell surface were excluded as explanations for the inhibitory activity of Cpn60.1. We therefore conclude that M. tuberculosis Cpn60.1 is an unusual protein with the ability to induce bipolar effects on human monocytes, which may help explain the pathology of granuloma formation in tuberculosis. We used microarrays to analyse the bipolar effectsof Cpn60.1 on human monocytes.
2015-02-17 | GSE18794 | GEO
Project description:Differences in the activity of monocytes/macrophages, important target cells of Mycobacterium tuberculosis, might influence tuberculosis progression. With the purpose of identifying candidate genes for tuberculosis susceptibility we infected with M. tuberculosis monocytes from both, healthy elders (a tuberculosis susceptibility group) and elderly tuberculosis patients, and performed a microarray experiment. We detected 78 differentially expressed transcripts and confirmed these results by quantitative PCR of selected genes. We found that monocytes from tuberculosis patients showed similar expression patterns of these genes regardless of whether they were obtained from younger or elder patients. Only one of the detected genes corresponded to a cytokine: IL-26, a member of the IL-10 cytokine family that we found downregulated in infected monocytes from tuberculosis patients. We have analyzed total RNA from Mycobacterium tuberculosis infected monocytes. We have isolated CD14+ cells (monocytes) from peripheral blood mononuclear cells by magnetic separation, and infected them for 4 days with 1 bacterium per monocyte. Blood donors were 7 elderly patients with pulmonary tuberculosis (average age: 83 years; sex: 3 men and 4 women) and 8 non-tuberculous volunteers (81 years, 6 men and 2 women).
2014-08-29 | E-GEOD-57736 | biostudies-arrayexpress
Project description:We propose a custom pipeline for combining data from experiments on multispecies studies tested on data from human and bovine peripheral blood mononuclear cellssamples, stimulated with Mycobacterium tuberculosis.
2023-09-15 | GSE218065 | GEO
Project description:Tuberculosis remains a major cause of death from an infectious disease worldwide, yet only 10% of people infected with Mycobacterium tuberculosis develop disease. Defining both necessary and sufficient immunologic determinants of protection remains a great scientific challenge. Analysis of peripheral blood gene expression profiles of active tuberculosis patients has identified correlates of risk for disease or pathogenesis. We sought to identify human potential candidate markers of host defense by studying gene expression profiles of macrophages, cells which, upon infection by M. tuberculosis, can mount an antimicrobial response. Weighted gene co-expression network analysis revealed an association between the cytokine, IL-32, and the vitamin D antimicrobial pathway in a network of IFN-γ and IL-15 induced ‘defense response’ genes. IL-32 was sufficient for induction of the vitamin D-dependent antimicrobial peptides, cathelicidin and DEFB4, and generation of antimicrobial activity in vitro, dependent on the presence of adequate 25-hydroxyvitamin D. The IL-15 induced ‘defense response’ macrophage gene network was integrated with ranked pairwise comparisons of gene expression from five different clinical data sets of latent vs. active tuberculosis or healthy controls, and a co-expression network derived from gene expression in patients with tuberculosis undergoing chemotherapy. Together, these analyses identified eight common genes, including IL-32, as molecular markers of latent tuberculosis and the IL-15 induced gene network. Inferring that maintaining M. tuberculosis in a latent state and preventing transition to active disease represents host resistance, we believe these results identify IL-32 as one functional marker and potential correlate of protection against active tuberculosis. Adherent peripheral blood monuclear cells were derived by Ficoll-Hypaque from the whole blood of four healthy donors. Cells adhered to tissue culture-treated plates for 2 h in 1% Fetal Bovine Serum (FBS) in RPMI. Cells were washed then stimulated with IL-10 (10ng/ml), IL-15 (10ng/ml) (R&D Systems), or IL-4 (1U/ml) in RPMI 1640 supplemented with 10% FBS at 37°C, 5% CO2. Cells were harvested at 6 h and 24 h after stimulation and monocytes purified by CD14 microbeads (Miltenyi Biotec) for a confirmed monocyte purity of at least 90%. RNA from purified monocytes extracted by Trizol and purified by Qiagen RNeasy Kit. RNA probe and microarray performed by UCLA Clinical Microarray Core using Ambion labeling kit and Affymetrix Human U133 Plus 2.0 array.
2014-08-20 | E-GEOD-59184 | biostudies-arrayexpress
Project description:This experiment explored the transcriptional response of human peripheral blood mononuclear cells (PBMC) isolated from BCG-vaccinated individuals following 6 days of in vitro stimulation with 2x10^5 cfu of different Bacillus Calmette-Guérin (BCG) strains or 100 ng/ml Mycobacterium tuberculosis-derived purified protein derivative (PPD). The BCG strains used were BCG Russia (Russian BCG-I sub-strain), BCG Japan (Tokyo 172 sub-strain), BCG Denmark (Danish 1331 sub-strain) & BCG Pasteur.
2022-01-26 | E-MTAB-11345 | biostudies-arrayexpress
Project description:human peripheral blood derived monocytes, LPS stimulation time-series
| PRJNA96153 | ENA
Project description:Mycobacterium tuberculosis (Mtb) causes a wide spectrum of human disease ranging from asymptomatic latent infection to severe host pathology, but it remains unclear how Mtb influence on leukocyte gene expression differs between disease states. We assessed mRNA expression in blood monocytes and granulocytes by next-generation sequencing in healthy controls and patients with active tuberculosis infection. We found genome-wide gene expression changes in granulocyte and monocyte which correlated with disease severity.
2021-11-05 | GSE126614 | GEO
Project description:Peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats by Ficoll-Paque centrifugation. Blood monocytes were then purified from PBMCs by positive selection with magnetic CD14 MicroBeads (Miltenyi Biotech). Pure monocytes were cultured for 5 days in RPMI 1640 (Invitrogen) supplemented with 10% heat-inactivated FCS (Dutscher), L-glutamine (Invitrogen), GM-CSF (20 ng/mL; Immunotools), and IL-4 (20 ng/mL; Immunotools). Cell cultures were fed every 2 days with complete medium supplemented with the cytokines previously mentioned. Before infection, we systematically checked the differentiation/activation status of the monocyte-derived DCs by flow cytometry, using antibodies against CD1a, CD14, CD83, and HLA-DR. Only samples presenting the expected phenotype for non-activated DCs – CD1a+, CD14-, CD83-, and HLA-DRlow – were used in downstream experiments. Dendritic cells (DCs) were infected with a Mycobacterium tuberculosis (MTB) strain expressing green-fluorescent protein (H37Rv) for 18 h at a multiplicity of infection of 1-to-1. Full details can be found in Barreiro et al. (2012).
2015-06-01 | GSE64177 | GEO
Project description:Current study will help us to elucidate the differential gene expression profiles of two different subsets of monocytes stimulated with gamma-M. tuberculosis CD14hiCD16- and CD14loCD16+ monocytes were isolated from human peripheral blood using magnetic beads for CD14 and CD16. 1.5 million cells were cultured in 1 ml of complete RPMI medium and stimulated with 10 microgram/ml of gamma-M. tuberculosis for 48 hours. After 48 hours cells were harvested and RNA was isolated from control and stimulated samples from CD14hiCD16- and CD14loCD16+ monocytes. So total we had 4 samples: 1. Control CD14hiCD16- 2. Stimulated CD14hiCD16- 3. Control CD14loCD16+ monocytes and 4. CD14loCD16+ monocytes. RNA from all these above 4 samples were sent to Phalanx Biotech Group for whole genome microarray.
2010-11-23 | E-GEOD-25505 | biostudies-arrayexpress
Project description:It is becoming increasingly apparent that Staphylococcus aureus are able to survive engulfment by macrophages, and that the intracellular environment of these cells, which is essential to innate host defenses against invading microorganisms, may in fact provide a refuge for staphylococcal survival and dissemination. Based on this, we postulated that S. aureus might induce cytoprotective mechanisms by changing gene expression profiles inside macrophages similar to obligate intracellular pathogens, such as Mycobacterium tuberculosis. In order to examine the effect of S. aureus on the macrophage transcriptome, we performed microarray expression analysis on human monocyte-derived macrophages treated with S. aureus. Experiment Overall Design: Human monocyte-derived macrophages (hMDMs) were separated from fractions of peripheral blood mononuclear cells (PBMCs) obtained from the blood of healthy donors. Control and S. aureus-exposed macrophages were incubated at 37C for 8, 24, or 48 hours
2009-05-02 | E-GEOD-13670 | biostudies-arrayexpress