scRNA-seq Reveals A Macrophage Subset That Provides A Splenic Replication Niche For Intracellular Salmonella

AbstractInteractions between intracellular bacteria and mononuclear phagocytes give rise to diverse cellular phenotypes that may determine the outcome of infection. Recent advances in single cell RNA-seq (scRNA-seq) have identified multiple subsets within the mononuclear population defined by unique molecular features, but the implications to their function during infection is unknown. Here, we applied high resolution kinetic analysis using microscopy, flow cytometry and scRNA-seq to survey the mononuclear niche of intracellular Salmonella Typhimurium (S.Tm) during early systemic infection in mice. We describe an eclipse like growth kinetics in the spleen, with a first phase of bacterial control mediated by tissue resident red pulp macrophages. A second phase involved bacterial growth mediated by intracellular replication within a macrophage population we termed CD9 macrophages, that originate from non-classical monocytes. Nr4a1e2−/− mice, specifically depleted of non-classical monocytes but not other mononuclear cells, are more resistant to S.Tm infection. Our study underscores a cell-type specific host-pathogen interaction that determines early infection growth dynamics and has implications to the infection outcome of the entire organism.

Download Full-text

A LINCS microenvironment perturbation resource for integrative assessment of ligand-mediated molecular and phenotypic responses

10.1101/2021.08.06.455429 ◽

2021 ◽

Author(s):

Sean M Gross ◽

Mark A Dane ◽

Rebecca L Smith ◽

Kaylyn Devlin ◽

Ian Mclean ◽

...

Keyword(s):

Mammary Epithelial Cells ◽

Molecular State ◽

Mammary Epithelial ◽

Systematic Assessment ◽

Extracellular Signals ◽

Molecular Features ◽

Integrative Assessment ◽

A Cell ◽

Cellular Phenotypes ◽

Phenotypic Responses

The phenotype of a cell and its underlying molecular state is strongly influenced by extracellular signals, including growth factors, hormones, and extracellular matrix. While these signals are normally tightly controlled, their dysregulation leads to phenotypic and molecular states associated with diverse diseases. To develop a detailed understanding of the linkage between molecular and phenotypic changes, we generated a comprehensive dataset that catalogs the transcriptional, proteomic, epigenomic and phenotypic responses of MCF10A mammary epithelial cells after exposure to the ligands EGF, HGF, OSM, IFNG, TGFB and BMP2. Systematic assessment of the molecular and cellular phenotypes induced by these ligands comprise the LINCS Microenvironment (ME) perturbation dataset, which has been curated and made publicly available for community-wide analysis and development of novel computational methods (synapse.org/LINCS_MCF10A). In illustrative analyses, we demonstrate how this dataset can be used to discover functionally related molecular features linked to specific cellular phenotypes.

Download Full-text

Deconvolution of monocyte responses in inflammatory bowel disease reveals an IL-1 cytokine network that regulates IL-23 in genetic and acquired IL-10 resistance

Gut ◽

10.1136/gutjnl-2020-321731 ◽

2020 ◽

pp. gutjnl-2020-321731

Author(s):

Dominik Aschenbrenner ◽

Maria Quaranta ◽

Soumya Banerjee ◽

Nicholas Ilott ◽

Joanneke Jansen ◽

...

Keyword(s):

Mononuclear Cells ◽

Personalised Medicine ◽

Mononuclear Phagocytes ◽

Cytokine Network ◽

Peripheral Blood Mononuclear ◽

Transcriptional Signature ◽

Inflammatory Monocytes ◽

Transcriptomic Signature ◽

Inflammatory Bowel

ObjectiveDysregulated immune responses are the cause of IBDs. Studies in mice and humans suggest a central role of interleukin (IL)-23-producing mononuclear phagocytes in disease pathogenesis. Mechanistic insights into the regulation of IL-23 are prerequisite for selective IL-23 targeting therapies as part of personalised medicine.DesignWe performed transcriptomic analysis to investigate IL-23 expression in human mononuclear phagocytes and peripheral blood mononuclear cells. We investigated the regulation of IL-23 expression and used single-cell RNA sequencing to derive a transcriptomic signature of hyperinflammatory monocytes. Using gene network correlation analysis, we deconvolved this signature into components associated with homeostasis and inflammation in patient biopsy samples.ResultsWe characterised monocyte subsets of healthy individuals and patients with IBD that express IL-23. We identified autosensing and paracrine sensing of IL-1α/IL-1β and IL-10 as key cytokines that control IL-23-producing monocytes. Whereas Mendelian genetic defects in IL-10 receptor signalling induced IL-23 secretion after lipopolysaccharide stimulation, whole bacteria exposure induced IL-23 production in controls via acquired IL-10 signalling resistance. We found a transcriptional signature of IL-23-producing inflammatory monocytes that predicted both disease and resistance to antitumour necrosis factor (TNF) therapy and differentiated that from an IL-23-associated lymphocyte differentiation signature that was present in homeostasis and in disease.ConclusionOur work identifies IL-10 and IL-1 as critical regulators of monocyte IL-23 production. We differentiate homeostatic IL-23 production from hyperinflammation-associated IL-23 production in patients with severe ulcerating active Crohn’s disease and anti-TNF treatment non-responsiveness. Altogether, we identify subgroups of patients with IBD that might benefit from IL-23p19 and/or IL-1α/IL-1β-targeting therapies upstream of IL-23.

Download Full-text

Glucocerebrosidase Activity is Reduced in Cryopreserved Parkinson’s Disease Patient Monocytes and Inversely Correlates with Motor Severity

Journal of Parkinson s Disease ◽

10.3233/jpd-202508 ◽

2021 ◽

pp. 1-9

Author(s):

Laura P. Hughes ◽

Marilia M.M. Pereira ◽

Deborah A. Hammond ◽

John B. Kwok ◽

Glenda M. Halliday ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Rating Scale ◽

Disease Patient ◽

Motor Symptom ◽

Blood Monocytes ◽

Peripheral Blood Monocytes ◽

Classical Monocytes

Background: Reduced activity of lysosomal glucocerebrosidase is found in brain tissue from Parkinson’s disease patients. Glucocerebrosidase is also highly expressed in peripheral blood monocytes where its activity is decreased in Parkinson’s disease patients, even in the absence of GBA mutation. Objective: To measure glucocerebrosidase activity in cryopreserved peripheral blood monocytes from 30 Parkinson’s disease patients and 30 matched controls and identify any clinical correlation with disease severity. Methods: Flow cytometry was used to measure lysosomal glucocerebrosidase activity in total, classical, intermediate, and non-classical monocytes. All participants underwent neurological examination and motor severity was assessed by the Movement Disorders Society Unified Parkinson’s Disease Rating Scale. Results: Glucocerebrosidase activity was significantly reduced in the total and classical monocyte populations from the Parkinson’s disease patients compared to controls. GCase activity in classical monocytes was inversely correlated to motor symptom severity. Conclusion: Significant differences in monocyte glucocerebrosidase activity can be detected in Parkinson’s disease patients using cryopreserved mononuclear cells and monocyte GCase activity correlated with motor features of disease. Being able to use cryopreserved cells will facilitate the larger multi-site trials needed to validate monocyte GCase activity as a Parkinson’s disease biomarker.

Download Full-text

Monocyte tissue factor induction by lipopolysaccharide (LPS): dependence on LPS-binding protein and CD14, and inhibition by a recombinant fragment of bactericidal/permeability-increasing protein

Blood ◽

10.1182/blood.v83.9.2516.2516 ◽

1994 ◽

Vol 83 (9) ◽

pp. 2516-2525 ◽

Cited By ~ 13

Author(s):

K Meszaros ◽

S Aberle ◽

R Dedrick ◽

R Machovich ◽

A Horwitz ◽

...

Keyword(s):

Tissue Factor ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Binding Protein ◽

Mononuclear Phagocytes ◽

Factor Vii ◽

Peripheral Blood Mononuclear ◽

Recombinant Fragment

Abstract Mononuclear phagocytes, stimulated by bacterial lipopolysaccharide (LPS), have been implicated in the activation of coagulation in sepsis and endotoxemia. In monocytes LPS induces the synthesis of tissue factor (TF) which, assembled with factor VII, initiates the blood coagulation cascades. In this study we investigated the mechanism of LPS recognition by monocytes, and the consequent expression of TF mRNA and TF activity. We also studied the inhibition of these effects of LPS by rBPI23, a 23-kD recombinant fragment of bactericidal/permeability increasing protein, which has been shown to antagonize LPS in vitro and in vivo. Human peripheral blood mononuclear cells, or monocytes isolated by adherence, were stimulated with Escherichia coli O113 LPS at physiologically relevant concentrations (> or = 10 pg/mL). The effect of LPS was dependent on the presence of the serum protein LBP (lipopolysaccharide-binding protein), as shown by the potentiating effect of human recombinant LBP or serum. Furthermore, recognition of low amounts of LPS by monocytes was also dependent on CD14 receptors, because monoclonal antibodies against CD14 greatly reduced the LPS sensitivity of monocytes in the presence of serum or rLBP. Induction of TF activity and mRNA expression by LPS were inhibited by rBPI23. The expression of tumor necrosis factor showed qualitatively similar changes. Considering the involvement of LPS-induced TF in the potentially lethal intravascular coagulation in sepsis, inhibition of TF induction by rBPI23 may be of therapeutic benefit.

Download Full-text

Loss of CXCR4 on non-classical monocytes in participants of the Women’s Interagency HIV Study (WIHS) with subclinical atherosclerosis

Cardiovascular Research ◽

10.1093/cvr/cvy292 ◽

2018 ◽

Vol 115 (6) ◽

pp. 1029-1040 ◽

Cited By ~ 5

Author(s):

Karin A L Mueller ◽

David B Hanna ◽

Erik Ehinger ◽

Xiaonan Xue ◽

Livia Baas ◽

...

Keyword(s):

Hiv Infection ◽

Mononuclear Cells ◽

Smoking Status ◽

Subclinical Atherosclerosis ◽

Cxcr4 Expression ◽

Monocyte Subset ◽

Surface Markers ◽

Peripheral Blood Mononuclear ◽

Cell Counts ◽

Classical Monocytes

AbstractAimsTo test whether human immunodeficiency virus (HIV) infection and subclinical cardiovascular disease (sCVD) are associated with expression of CXCR4 and other surface markers on classical, intermediate, and non-classical monocytes in women.Methods and resultssCVD was defined as presence of atherosclerotic lesions in the carotid artery in 92 participants of the Women’s Interagency HIV Study (WIHS). Participants were stratified into four sets (n = 23 each) by HIV and sCVD status (HIV−/sCVD−, HIV−/sCVD+, HIV+/sCVD−, and HIV+/sCVD+) matched by age, race/ethnicity, and smoking status. Three subsets of monocytes were determined from archived peripheral blood mononuclear cells. Flow cytometry was used to count and phenotype surface markers. We tested for differences by HIV and sCVD status accounting for multiple comparisons. We found no differences in monocyte subset size among the four groups. Expression of seven surface markers differed significantly across the three monocyte subsets. CXCR4 expression [median fluorescence intensity (MFI)] in non-classical monocytes was highest among HIV−/CVD− [628, interquartile range (IQR) (295–1389)], followed by HIV+/CVD− [486, IQR (248–699)], HIV−/CVD+ (398, IQR (89–901)), and lowest in HIV+/CVD+ women [226, IQR (73–519)), P = 0.006 in ANOVA. After accounting for multiple comparison (Tukey) the difference between HIV−/CVD− vs. HIV+/CVD+ remained significant with P = 0.005 (HIV−/CVD− vs. HIV+/CVD− P = 0.04, HIV−/CVD− vs. HIV−/CVD+ P = 0.06, HIV+/CVD+ vs. HIV+/CVD− P = 0.88, HIV+/CVD+ vs. HIV−/CVD+ P = 0.81, HIV+/CVD− vs. HIV−/CVD+, P = 0.99). All pairwise comparisons with HIV−/CVD− were individually significant (P = 0.050 vs. HIV−/CVD+, P = 0.028 vs. HIV+/CVD−, P = 0.009 vs. HIV+/CVD+). CXCR4 expression on non-classical monocytes was significantly higher in CVD− (501.5, IQR (249.5–887.3)) vs. CVD+ (297, IQR (81.75–626.8) individuals (P = 0.028, n = 46 per group). CXCR4 expression on non-classical monocytes significantly correlated with cardiovascular and HIV−related risk factors including systolic blood pressure, platelet and T cell counts along with duration of antiretroviral therapy (P < 0.05). In regression analyses, adjusted for education level, study site, and injection drug use, presence of HIV infection and sCVD remained significantly associated with lower CXCR4 expression on non-classical monocytes (P = 0.003), but did not differ in classical or intermediate monocytes.ConclusionCXCR4 expression in non-classical monocytes was significantly lower among women with both HIV infection and sCVD, suggesting a potential atheroprotective role of CXCR4 in non-classical monocytes.

Download Full-text

Proteomic analysis reveals presence of platelet microparticles in endothelial progenitor cell cultures

Blood ◽

10.1182/blood-2009-02-205930 ◽

2009 ◽

Vol 114 (3) ◽

pp. 723-732 ◽

Cited By ~ 199

Author(s):

Marianna Prokopi ◽

Giordano Pula ◽

Ursula Mayr ◽

Cécile Devue ◽

Joy Gallagher ◽

...

Keyword(s):

Clinical Trials ◽

Mononuclear Cells ◽

Lectin Binding ◽

Large Population ◽

Protein Composition ◽

Population Based ◽

Endothelial Progenitor ◽

Cardiovascular Research ◽

Proteomic Approach ◽

Cellular Phenotypes

Abstract The concept of endothelial progenitor cells (EPCs) has attracted considerable interest in cardiovascular research, but despite a decade of research there are still no specific markers for EPCs and results from clinical trials remain controversial. Using liquid chromatography–tandem mass spectrometry, we analyzed the protein composition of microparticles (MPs) originating from the cell surface of EPC cultures. Our data revealed that the conventional methods for isolating mononuclear cells lead to a contamination with platelet proteins. Notably, platelets readily disintegrate into platelet MPs. These platelet MPs are taken up by the mononuclear cell population, which acquires “endothelial” characteristics (CD31, von Willebrand factor [VWF], lectin-binding), and angiogenic properties. In a large population-based study (n = 526), platelets emerged as a positive predictor for the number of colony-forming units and early outgrowth EPCs. Our study provides the first evidence that the cell type consistent with current definitions of an EPC phenotype may arise from an uptake of platelet MPs by mononuclear cells resulting in a gross misinterpretation of their cellular progeny. These findings demonstrate the advantage of using an unbiased proteomic approach to assess cellular phenotypes and advise caution in attributing the benefits in clinical trials using unselected bone marrow mononuclear cells (BMCs) to stem cell-mediated repair.

Download Full-text

Genome-Wide DNA Methylation Profiles of Phlegm-Dampness Constitution

Cellular Physiology and Biochemistry ◽

10.1159/000487976 ◽

2018 ◽

Vol 45 (5) ◽

pp. 1999-2008 ◽

Cited By ~ 3

Author(s):

Haiqiang Yao ◽

Shanlan Mo ◽

Ji Wang ◽

Yingshuai Li ◽

Chong-Zhi Wang ◽

...

Keyword(s):

Dna Methylation ◽

Metabolic Disorders ◽

Mononuclear Cells ◽

Metabolic Diseases ◽

Body Type ◽

Peripheral Blood Mononuclear ◽

Molecular Features ◽

Genome Wide ◽

A Genome ◽

Differentially Methylated Genes

Background/Aims: Metabolic diseases are leading health concerns in today’s global society. In traditional Chinese medicine (TCM), one body type studied is the phlegm-dampness constitution (PC), which predisposes individuals to complex metabolic disorders. Genomic studies have revealed the potential metabolic disorders and the molecular features of PC. The role of epigenetics in the regulation of PC, however, is unknown. Methods: We analyzed a genome-wide DNA methylation in 12 volunteers using Illumina Infinium Human Methylation450 BeadChip on peripheral blood mononuclear cells (PBMCs). Eight volunteers had PC and 4 had balanced constitutions. Results: Methylation data indicated a genome-scale hyper-methylation pattern in PC. We located 288 differentially methylated probes (DMPs). A total of 256 genes were mapped, and some of these were metabolic-related. SQSTM1, DLGAP2 and DAB1 indicated diabetes mellitus; HOXC4 and SMPD3, obesity; and GRWD1 and ATP10A, insulin resistance. According to Ingenuity Pathway Analysis (IPA), differentially methylated genes were abundant in multiple metabolic pathways. Conclusion: Our results suggest the potential risk for metabolic disorders in individuals with PC. We also explain the clinical characteristics of PC with DNA methylation features.

Download Full-text

Overview: Autologous Peripheral Blood Mononuclear Cells

Journal of Stem Cell Research and Transplantation ◽

10.26420/jstemcellrestransplant.2021.1034 ◽

2021 ◽

Vol 8 (1) ◽

Author(s):

Petrungaro A ◽

◽

Scudo F ◽

Quartarone E ◽

◽

...

Keyword(s):

Tissue Regeneration ◽

Tissue Repair ◽

Mononuclear Cells ◽

Mononuclear Phagocyte ◽

Tendon Tissue ◽

Central Importance ◽

Peripheral Blood Mononuclear ◽

Avant Garde ◽

Blood Mononuclear Cells ◽

A Cell

The implant of autologous mononuclear cells is an avant-garde method based on the use of a cell population within our body to regenerate tissues that have been damaged by various pathological events. The biological assumption is the richness and complexity of biochemical and cellular phenomena inherent in both organism response to damage and tissue regeneration. The key role is played by the mononuclear phagocyte system which regulates and modulates the activity of mesenchymal stem cells capable of differentiating and providing tissue repair. This system does not only have a traditional “scavenger” function within the immune system, but it is also of central importance in the modulation and activation of the response to tissue damage, be it traumatic, surgical, or degenerative. In this review we summarize the main features of this method and the most common uses in clinical practice where the interest is growing considering both the powerful, rapid and documented regenerative response of the various “target” tissues: vascular, cartilage, bone, muscle and tendon tissue.

Download Full-text

Increased in vitro and in vivo generation of procoagulant activity (tissue factor) by mononuclear phagocytes after intralipid infusion in rabbits

Blood ◽

10.1182/blood.v65.6.1391.bloodjournal6561391 ◽

1985 ◽

Vol 65 (6) ◽

pp. 1391-1395 ◽

Cited By ~ 15

Author(s):

P Montemurro ◽

A Lattanzio ◽

G Chetta ◽

L Lupo ◽

L Caputi-Iambrenghi ◽

...

Keyword(s):

Tissue Factor ◽

Mononuclear Cells ◽

Mononuclear Phagocyte ◽

Mononuclear Phagocytes ◽

Procoagulant Activity ◽

Sterile Saline ◽

Functional Changes ◽

Before And After

Abstract Intralipid, a fat emulsion widely used in parenteral nutrition, can produce marked functional changes of the mononuclear phagocyte system. We investigated the effect of Intralipid administration on the generation of procoagulant activity by rabbit mononuclear phagocytes. Two groups of ten rabbits given either a single infusion of Intralipid 10% or a similar volume of sterile saline were studied before and after infusion. Procoagulant activity was measured on isolated blood mononuclear cells after incubation with and without endotoxin, using a one-stage clotting assay. Cells from animals infused with Intralipid produced significantly more procoagulant activity than controls (P less than .01). Results were similar when freshly collected whole blood was incubated with and without endotoxin, and procoagulant activity was measured on subsequently isolated mononuclear cells (P less than .01). In addition, when rabbits were given a single injection of endotoxin, blood and spleen mononuclear cells harvested 50 to 60 minutes after the injection from animals pretreated with Intralipid expressed five to seven times more procoagulant activity than did cells from animals pretreated with saline. In all instances, procoagulant activity was identified as tissue factor. These findings suggest that Intralipid may cause functional changes in mononuclear phagocytes, resulting in increased production of tissue factor on incubation in short-term culture in vitro and in response to endotoxin in vivo.

Download Full-text

In vivo production of interleukin-5, granulocyte-macrophage colony- stimulating factor, macrophages colony-stimulating factor, and interleukin-6 during intravenous administration of high-dose interleukin-2 in cancer patients [see comments]

Blood ◽

10.1182/blood.v78.8.1981.1981 ◽

1991 ◽

Vol 78 (8) ◽

pp. 1981-1987 ◽

Cited By ~ 29

Author(s):

MR Schaafsma ◽

JH Falkenburg ◽

JE Landegent ◽

N Duinkerken ◽

S Osanto ◽

...

Keyword(s):

Cancer Patients ◽

Mononuclear Cells ◽

Bone Marrow Cells ◽

Interleukin 2 ◽

Mononuclear Phagocytes ◽

High Dose ◽

Colony Stimulating Factor ◽

Gm Csf ◽

Stimulating Factor

Abstract Recombinant human interleukin-2 (IL-2), administered to cancer patients by continuous intravenous (IV) infusion (3 x 10(6) U/m2/d), was found to induce the in vivo production of colony-stimulating factors (CSF). Plasma obtained from patients during IL-2 treatment stimulated in vitro colony formation of normal human bone marrow cells, depleted of mononuclear phagocytes and T lymphocytes. This colony-stimulating activity (CSA) was identified as IL-5, granulocyte-macrophage CSF (GM- CSF), and macrophage CSF (M-CSF), by the ability of specific antibodies against these factors to neutralize their effects. The presence of IL-2- induced GM-CSF and M-CSF was also demonstrated by specific radioimmunoassays. During IL-2 treatment, plasma also contained detectable levels of IL-6, which was measured in a bioassay. Using a cDNA-polymerase chain reaction (PCR) with specific primer sets for the various CSF, we showed that IL-2 treatment induced the expression of mRNA for M-CSF, GM-CSF, IL-3, and IL-5, but not for granulocyte CSF (G- CSF) in peripheral blood mononuclear cells, suggesting differential expression of CSF in vivo in response to IL-2. Furthermore, no negative regulators of hematopoiesis, such as interferon gamma (IFN-gamma) or tumor necrosis factor-alpha (TNF-alpha), were found in plasma. These data illustrate that in vivo administration of high-dose IL-2 may result in a stimulatory effect on hematopoiesis. The induction of detectable levels of IL-5 and GM-CSF in the circulation may explain the eosinophilia and neutrophilia observed in these patients.

Download Full-text