Journal: medRxiv

Article Title: M2 monocyte polarization in dialyzed patients is associated with increased levels of M-CSF and myeloperoxidase-associated oxidative stress: preliminary results

doi: 10.1101/2020.05.07.20094011

Figure Lengend Snippet: M1 and M2 monocytes in the blood of HD patients (n = 27) and healthy subjects (n = 23) were analyzed by flow cytometry. (A) Quantification of classical, intermediate and non-classical monocytes. Quantification of M1 monocytes assessed by the positive expression of CD86 and CCR2 (B) and of M2 monocytes assessed by the positive expression of CD206, CXCR3 and CD163 (C). Quantification of M1 (D) and M2 (E) monocytes in classical, intermediate and non-classical monocyte subsets. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: 100 μL of total blood from patients were incubated for 15 minutes at RT with PE mouse anti-human CD14 and V500 mouse anti-human CD16 antibodies (Becton Dickinson, Franklin Lakes, NJ, USA) as well as with mouse anti-human CD86-FITC and anti-human CCR2-APC monoclonal antibodies (Miltenyi Biotec, Bergisch Gladbach, Germany) for determining the M1 polarization or with mouse anti-human CD206-FITC, anti-human CXCR3-APC and anti-human CD163-VioBlue monoclonal antibodies (Miltenyi Biotec, Bergisch Gladbach, Germany) for the M2 polarization.

Techniques: Flow Cytometry, Expressing

Journal: BMC Veterinary Research

Article Title: Accumulation profiles of PrP Sc in hemal nodes of naturally and experimentally scrapie-infected sheep

doi: 10.1186/1746-6148-9-82

Figure Lengend Snippet: mAbs used to identify leukocyte subsets in hemal and retropharyngeal lymph nodes during immunohistochemistry study

Article Snippet: Mac , CD163 , MCA1853 , mouse IgG1 , 33.3 , AbD Serotec.

Techniques: Immunohistochemistry, Marker, Clone Assay

Journal: Glia

Article Title: The Transcription Factor p53 Influences Microglial Activation Phenotype

doi: 10.1002/glia.21178

Figure Lengend Snippet: Separate populations of microglia demonstrate immunoreactivity for a marker of alternatively activated and deactivated macrophages, CD163 or p53 activation in HAND cases. A) Human cortical tissue sections immunolabeled for p53 (brown) and CD68 (purple) reveal that a portion of microglia are immunoreactive for both markers. Open arrows point to double labeled cells and closed arrows identify CD68 single labeled cells. B) A section adjacent to the one in A is shown labeled with antibodies to p53 (brown) and CD163 (purple). Closed arrows point to CD163 labeled microglia. C) Quantification of double labeled microglia in adjacent sections from twelve HAND patients (*=p<0.0001 using a paired t-test).

Article Snippet: Co-labeling for p53 and CD163 was performed by adapting a previously published protocol ( Fischer-Smith et al. 2008 ) using a monoclonal antibody to CD163 (Vector 1:50) and co-labeling with antibodies to p53 using the same method as that developed to double label for p53 and CD68.

Techniques: Marker, Activation Assay, Immunolabeling, Labeling

Journal: International Journal of Molecular Sciences

Article Title: Multiple Immunostainings with Different Epitope Retrievals—The FOLGAS Protocol

doi: 10.3390/ijms23010223

Figure Lengend Snippet: Secondary antibodies, chromogens, fluorophores and metals. Except DAPI, the fluorophores not directly coupled to antibodies are linked to streptavidin for binding of biotinylated antibodies. Abbreviations: RTU: ready to use, AP: alkaline phosphatase, HRP: horseradish peroxidase. Tb: Terbium, Er: Erbium.

Article Snippet: , 170Er (conjugated to CD163 clone EDHu-1) , , - , Fluidigm (SKU 201300).

Techniques: Binding Assay

Journal: Journal of Clinical Medicine

Article Title: Compartment-Specific Differences in the Activation of Monocyte Subpopulations Are Not Affected by Nitric Oxide and Glucocorticoid Treatment in a Model of Resuscitated Porcine Endotoxemic Shock

doi: 10.3390/jcm11092641

Figure Lengend Snippet: Endotoxin-induced changes in the circulating cell counts and phenotypes. Time-course changes in: ( A ) White blood count (WBC), ( B ) Circulating mononuclear cells (MNCs), and ( C ) CD3 + T-cells. ( D ) Representative dot plots of the flow cytometric analysis of blood monocytes; first, all cells were gated, and then the gate for the MNCs was applied and this population was analyzed for the expression of CD163 and CD14 before the infusion of endotoxin, 4 h and 20 h after the start of endotoxemia. The frequency of: ( E ) CD14 + CD163 − monocytes, ( F ) CD14 + CD163 + monocytes, and ( G ) CD14 − CD163 + monocytes is shown. Mean values ± SD are presented. Baseline, T4 h T8 h n = 9/group, T20 h n = 6/group. The statistical significance of differences between values at different timepoints for each group: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. The statistical significance of differences between groups at given timepoints: ## p < 0.01. FSC forward scatter, SSC side scatter.

Article Snippet: The following antibodies for immunophenotyping of the monocytes were used: anti-CD14-PE.Cy7 (clone M5E2, BioLegend, San Diego, CA, USA), anti-CD163 (2A10/11, Bio-Rad (AbD Serotec), Hercules, CA, USA) labeled with Zenon AlexaFluor 647 mouse IgG1 (Thermofisher), and anti-SLA-DR-FITC (2E9/13, Bio-Rad (AbD Serotec)).

Techniques: Expressing

Journal: Journal of Clinical Medicine

Article Title: Compartment-Specific Differences in the Activation of Monocyte Subpopulations Are Not Affected by Nitric Oxide and Glucocorticoid Treatment in a Model of Resuscitated Porcine Endotoxemic Shock

doi: 10.3390/jcm11092641

Figure Lengend Snippet: Effects of endotoxemia on the activation status of circulating monocytes. The kinetics of SLA-DR expression on subpopulations of circulating blood monocytes with representative histograms from flow cytometry analysis is shown: ( A ) CD14 + CD163 − monocytes, ( B ) CD14 + CD163 + monocytes, and ( C ) CD14 − CD163 + monocytes. The mitochondrial membrane potential (Δφ) was analyzed using JC-1 staining followed by flow cytometric analysis on: ( D ) CD14 + CD163 + monocytes, ( E ) CD14 + CD163 + monocytes, and ( F ) CD14 + CD163 − monocytes. Mean values ±SD are presented. Baseline, T4 h T8 h n = 9/group, T20 h n = 6/group. The statistical significance of differences between values at different timepoints for each group: ** p < 0.01, *** p < 0.001, **** p < 0.0001. SLA-DR swine leukocyte antigen-DR.

Article Snippet: The following antibodies for immunophenotyping of the monocytes were used: anti-CD14-PE.Cy7 (clone M5E2, BioLegend, San Diego, CA, USA), anti-CD163 (2A10/11, Bio-Rad (AbD Serotec), Hercules, CA, USA) labeled with Zenon AlexaFluor 647 mouse IgG1 (Thermofisher), and anti-SLA-DR-FITC (2E9/13, Bio-Rad (AbD Serotec)).

Techniques: Activation Assay, Expressing, Flow Cytometry, Staining

Journal: Journal of Clinical Medicine

Article Title: Compartment-Specific Differences in the Activation of Monocyte Subpopulations Are Not Affected by Nitric Oxide and Glucocorticoid Treatment in a Model of Resuscitated Porcine Endotoxemic Shock

doi: 10.3390/jcm11092641

Figure Lengend Snippet: Compartment-specific changes in the activation status of monocytes following 20 h of endotoxemic shock. The expression of SLA-DR antigen was evaluated 20 h after the onset of endotoxemic shock in cells from the bronchoalveolar lavage fluid (BAL), peripheral blood (PB), and bone marrow (BM). The SLA-DR antigen was analyzed in: ( A ) CD14 + CD163 − monocytes, ( B ) CD14 + CD163 + monocytes, and ( C ) CD14 − CD163 + monocytes. The mitochondria membrane potential (Δφ) was also analyzed in these three compartments in: ( D ) CD14 + CD163 − monocytes, ( E ) CD14 + CD163 + monocytes, and ( F ) CD14 − CD163 + monocytes. Mean values ± SD are presented. n = 6/group. The statistical significance of differences between values for BAL vs. PB or BM in each experimental group: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Significant differences between PB and BM §§§§ p < 0.0001. No differences were noted between control and treated group.

Article Snippet: The following antibodies for immunophenotyping of the monocytes were used: anti-CD14-PE.Cy7 (clone M5E2, BioLegend, San Diego, CA, USA), anti-CD163 (2A10/11, Bio-Rad (AbD Serotec), Hercules, CA, USA) labeled with Zenon AlexaFluor 647 mouse IgG1 (Thermofisher), and anti-SLA-DR-FITC (2E9/13, Bio-Rad (AbD Serotec)).

Techniques: Activation Assay, Expressing