scholarly journals Single-cell analysis reveals new subset markers of murine peritoneal macrophages and highlights macrophage dynamics upon Staphylococcus aureus peritonitis

2016 ◽  
Vol 22 (5) ◽  
pp. 382-392 ◽  
Author(s):  
Solène Accarias ◽  
Clémence Genthon ◽  
David Rengel ◽  
Séverine Boullier ◽  
Gilles Foucras ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Flow Cytometry ◽  
Single Cell ◽  
Single Cell Analysis ◽  
Immune Surveillance ◽  
Peritoneal Macrophages ◽  
Aureus Infection ◽  
Inflammatory Monocytes ◽  
Peritonitis Model

Resident macrophages play a central role in maintaining tissue homeostasis and immune surveillance. Here, we used single cell-based qPCR coupled with flow cytometry analysis to further define the phenotypes of large and small resident peritoneal macrophages (LPMs and SPMs, respectively) in mice. We demonstrated that the expression of Cxcl13, IfngR1, Fizz-1 and Mrc-1 clearly distinguished between LPMs and SPMs subsets. Using these markers, the dynamics of peritoneal macrophages in a Staphylococcus aureus-induced peritonitis model were analyzed. We found that S. aureus infection triggers a massive macrophage disappearance reaction in both subsets. Thereafter, inflammatory monocytes rapidly infiltrated the cavity and differentiated to replenish the SPMs. Although phenotypically indistinguishable from resident SPMs by flow cytometry, newly recruited SPMs had a different pattern of gene expression dominated by M2 markers combined with M1 associated features (inos expression). Interestingly, S. aureus elicited SPMs showed a robust expression of Cxcl13, suggesting that these cells may endorse the role of depleted LPMs and contribute to restoring peritoneal homeostasis. These data provide information on both resident and recruited macrophages dynamics upon S. aureus infection and demonstrate that single-cell phenotyping is a promising and highly valuable approach to unraveling macrophage diversity and plasticity.

scholarly journals Role of Purine Biosynthesis in Persistent Methicillin-Resistant Staphylococcus aureus Infection

2018 ◽  
Vol 218 (9) ◽  
pp. 1367-1377 ◽  
Author(s):  
Liang Li ◽  
Wessam Abdelhady ◽  
Niles P Donegan ◽  
Kati Seidl ◽  
Ambrose Cheung ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Purine Biosynthesis ◽  
Methicillin Resistant ◽  
Aureus Infection

Electroporative Flow Cytometry for Single-Cell Analysis

2010 ◽  
pp. 125-142
Author(s):  
Chang Lu ◽  
Jun Wang ◽  
Ning Bao ◽  
Hsiang-Yu Wang
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
Single Cell Analysis ◽  
Cell Analysis

Single Cell Analysis of Microalgae and Associated Bacteria Flora by Using Flow Cytometry

2021 ◽  
Vol 26 (6) ◽  
pp. 898-909
Author(s):  
Fabrizio Di Caprio ◽  
Simone Posani ◽  
Pietro Altimari ◽  
Alessandro Concas ◽  
Francesca Pagnanelli
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Associated Bacteria

Role of microRNAs in Staphylococcus aureus infection: Potential biomarkers and mechanism

IUBMB Life ◽  
2020 ◽  
Vol 72 (9) ◽  
pp. 1856-1869 ◽  
Author(s):  
Rasoul Mirzaei ◽  
Rokhsareh Mohammadzadeh ◽  
Hamed Mirzaei ◽  
Mohammad Sholeh ◽  
Sajad Karampoor ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Aureus Infection ◽  
Potential Biomarkers

Single‐Cell Analysis of Cytokine mRNA and Protein Expression by Flow Cytometry

2020 ◽  
Vol 92 (1) ◽  
Author(s):  
Rubina Pal ◽  
Jayne Schaubhut ◽  
Darcey Clark ◽  
Lynette Brown ◽  
Jennifer J. Stewart
Keyword(s):  
Flow Cytometry ◽  
Protein Expression ◽  
Single Cell ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Cytokine Mrna ◽  
Mrna And Protein Expression

scholarly journals Down-regulation of mannosyl receptor-mediated endocytosis and antigen F4/80 in bacillus calmette-guerin-activated mouse macrophages. Role of T lymphocytes and lymphokines

1982 ◽  
Vol 155 (6) ◽  
pp. 1623-1637 ◽  
Author(s):  
AB Ezekowitz ◽  
S Gordon
Keyword(s):  
Plasma Membrane ◽  
Single Cell Analysis ◽  
Target Population ◽  
Peritoneal Macrophages ◽  
Membrane Receptors ◽  
Lymphocyte Function ◽  
Bacillus Calmette Guerin ◽  
Down Regulation ◽  
Receptor Mediated Endocytosis

Bacillus Calmette-Guerin (BCG) infection alters the surface and endocytic properties of mouse peritoneal macrophages (PM) compared with thioglycollate- elicited (TPM) or resident PM (RPM). Expression of Ia antigen (Ag) is enhanced up to fourfold, but plasma membrane receptors that mediate binding and uptake of mannosyl/fucosyl-terminated glycoconjugates (MFR), Fc receptors, and the macrophage (mφ)-specific Ag F4/80 are reduced by 50-80 percent. Levels of Mac-1 remain relatively stable. These changes are accompanied by enhanced secretion of O(2)(-), after further stimulation with phorbyl myristate acetate, and of plasminogen activator. Both these products are released by TPM, but not RPM. The characteristic surface phenotype of BCG-PM can also be induced by injection of C. parvum, another mφ- activating agent, but not by thioglycollate broth, lipopolysaccharide, or proteose peptone. Purified protein derivative (PPD) and N-acetylmuramyl-L- alanyl-D-isoglutamine. 2H(2)0 are soluble agents with partial activity. Alteration of mφ markers by BCG infection depends on T lymphocyte function, although studies with nude mice indicate that other pathways may also serve to modify the surface of the mφ. Mφ from uninfected animals displayed all markers of activation after adoptive transfer of specifically-sensitised lymphocytes with PPD, intraperitoneally, or after co- cultivation. Treatment of primed lymphocytes with anti-Thy-1 antibody and complement ablated this effect. Lymphokines obtaned by Ag or mitogen stimulation induced similar changes in TPM and RPM. Mannose-specific endocytosis decayed rapidly, time 1/2 approximately equal to 16 h and stabilized at approximately 25 percent of control values. Single-cell analysis showed that residual MFR activity was uniform in the target population. Loss of Ag F4/80 after activation by lymphocyte and PPD was less marked than after infection (35 percent vs 80 percent), unlike MFR activity, which declined to a similar extent. Induction of mφ Ia by lymphokine reached a peak after 2-3 d and was lost within 2 d of its removal. Recovery of MFR and F4/80 was incomplete under these conditions. These studies establish that activated mφ known to display enhanced antimicrobial/anticellular activity express markedly different surface properties distinct from elicited or resident cells. The role of antigen- stimulated T cell products in regulating mφ function is confirmed, and down-regulation of mannosyl-receptor-mediated endocytosis provides a sensitive, quantitative, and cell-specific new marker to study their properties and mechanism of action. Extensive, but selective remodeling of mφ plasma membrane structure could play an important role in controlling recognition and effector mechanisms of the activated mφ.

scholarly journals Using Flow Cytometry and Multistage Machine Learning to Discover Label-Free Signatures of Algal Lipid Accumulation

2018 ◽  
Author(s):  
Mohammad Tanhaemami ◽  
Elaheh Alizadeh ◽  
Claire Sanders ◽  
Babetta L. Marrone ◽  
Brian Munsky’
Keyword(s):  
Machine Learning ◽  
Flow Cytometry ◽  
Single Cell ◽  
Lipid Accumulation ◽  
Time Course ◽  
Fluorescent Dyes ◽  
Single Cell Analysis ◽  
Learning Approaches ◽  
Label Free ◽  
Unlabeled Cell

Abstract—Most applications of flow cytometry or cell sorting rely on the conjugation of fluorescent dyes to specific biomarkers. However, labeled biomarkers are not always available, they can be costly, and they may disrupt natural cell behavior. Label-free quantification based upon machine learning approaches could help correct these issues, but label replacement strategies can be very difficult to discover when applied labels or other modifications in measurements inadvertently modify intrinsic cell properties. Here we demonstrate a new, but simple approach based upon feature selection and linear regression analyses to integrate statistical information collected from both labeled and unlabeled cell populations and to identify models for accurate label-free single-cell quantification. We verify the method’s accuracy to predict lipid content in algal cells(Picochlorum soloecismus)during a nitrogen starvation and lipid accumulation time course. Our general approach is expected to improve label-free single-cell analysis for other organisms or pathways, where biomarkers are inconvenient, expensive, or disruptive to downstream cellular processes.

scholarly journals Role of Calcium Signaling Pathway-Related Gene Regulatory Networks in Ischemic Stroke Based on Multiple WGCNA and Single-Cell Analysis

2021 ◽  
Vol 2021 ◽  
pp. 1-35
Author(s):  
Weiwei Lin ◽  
Yangxin Wang ◽  
Yisheng Chen ◽  
Qiangwei Wang ◽  
Zhaowen Gu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Calcium Signaling ◽  
Single Cell ◽  
Signaling Pathway ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Kegg Pathways ◽  
Calcium Signaling Pathway ◽  
Core Genes

Background. This study is aimed at investigating the changes in relevant pathways and the differential expression of related gene expression after ischemic stroke (IS) at the single-cell level using multiple weighted gene coexpression network analysis (WGCNA) and single-cell analysis. Methods. The transcriptome expression datasets of IS samples and single-cell RNA sequencing (scRNA-seq) profiles of cerebrovascular tissues were obtained by searching the Gene Expression Omnibus (GEO) database. First, gene pathway scoring was calculated via gene set variation analysis (GSVA) and was imported into multiple WGCNA to acquire key pathways and pathway-related hub genes. Furthermore, SCENIC was used to identify transcription factors (TFs) regulating these core genes using scRNA-seq data. Finally, the pseudotemporal trajectory analysis was used to analyse the role of these TFs on various cell types under hypoxic and normoxic conditions. Results. The scores of 186 KEGG pathways were obtained via GSVA using microarray expression profiles of 40 specimens. WGCNA of the KEGG pathways revealed the two following pathways: calcium signaling pathway and neuroactive ligand-receptor interaction pathways. Subsequently, WGCNA of the gene expression matrix of the samples revealed the calcium signaling pathway-related genes (AC079305.10, BCL10, BCL2A1, BRE-AS1, DYNLL2, EREG, and PTGS2) that were identified as core genes via correlation analysis. Furthermore, SCENIC and pseudotemporal analysis revealed JUN, IRF9, ETV5, and PPARA score gene-related TFs. Jun was found to be associated with hypoxia in endothelial cells, whereas Irf9 and Etv5 were identified as astrocyte-specific TFs associated with oxygen concentration in the mouse cerebral cortex. Conclusions. Calcium signaling pathway-related genes (AC079305.10, BCL10, BCL2A1, BRE-AS1, DYNLL2, EREG, and PTGS2) and TFs (JUN, IRF9, ETV5, and PPARA) were identified to play a key role in IS. This study provides a new perspective and basis for investigating the pathogenesis of IS and developing new therapeutic approaches.

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