Journal: Nephrology Dialysis Transplantation

Article Title: Distinct immunologic effects of different intravenous iron preparations on monocytes

doi: 10.1093/ndt/gft524

Figure Lengend Snippet: Expression of CD14, CD16 and CD86 on circulating mature classical, intermediate and nonclassical monocytes from control subjects

Article Snippet: The following antibodies were used: anti-CD14 PerCP (Mφ9), anti-CD16 PeCy7 (3G8) and anti-CD195 APC (2D7/CCR5) and anti-TNF FITC (MAb11) (BD Biosciences, Heidelberg, Germany), anti-CD86 PE (HA5.2B7) (Beckman-Coulter, Krefeld, Germany), anti-CX 3 CR1 FITC (2A9-1) (Biozol, Eching, Germany), and anti-IL-1β Alexa Fluor® 647 (JK1B-1) (Biolegend, Fell, Germany).

Techniques: Expressing

Journal: Nephrology Dialysis Transplantation

Article Title: Distinct immunologic effects of different intravenous iron preparations on monocytes

doi: 10.1093/ndt/gft524

Figure Lengend Snippet: Expression of CD14, CD16 and CD86 on classical and intermediate monocytes after differentiation from haematopoietic stem cells from control subjects

Article Snippet: The following antibodies were used: anti-CD14 PerCP (Mφ9), anti-CD16 PeCy7 (3G8) and anti-CD195 APC (2D7/CCR5) and anti-TNF FITC (MAb11) (BD Biosciences, Heidelberg, Germany), anti-CD86 PE (HA5.2B7) (Beckman-Coulter, Krefeld, Germany), anti-CX 3 CR1 FITC (2A9-1) (Biozol, Eching, Germany), and anti-IL-1β Alexa Fluor® 647 (JK1B-1) (Biolegend, Fell, Germany).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Monocyte Subset Dynamics in Human Atherosclerosis Can Be Profiled with Magnetic Nano-Sensors

doi: 10.1371/journal.pone.0005663

Figure Lengend Snippet: A. Flow cytometry dot plots show forward scatter (FSC) versus side scatter (SSC) of mononuclear cells obtained from fresh blood. A monocyte gate is drawn and monocyte subsets are identified according to their CD14 and CD16 expression profile. B. Histograms depict MCSFR expression of CD16 lo monocytes, CD16 hi monocytes and other cells (mostly lymphocytes). C. Table summarizes relative expression profiles of selected markers for CD16 lo , CD16 hi monocytes and other cells. D. Representative histograms and H&E cytospin preparations show CD68 expression and morphology of CD16 lo , CD16 hi monocytes and other cells freshly isolated (□) or after in vitro culture for 6 days with LPS/IFNγ. E. Bar graph depicts ex vivo phagocytosis of fluorescently labeled latex beads in CD16 lo , CD16 hi monocytes and other cells (n = 4).

Article Snippet: Cell suspensions were stained with the following antibodies (all from BD Bioscience, unless otherwise stated) at a final concentration of 1∶100: CD11b-APC-Cy7/ICRF44, CD14-PE/M5E2, CD16 PE-Cy7/3G8, CCR2-Alexa-647/48607, CX 3 CR1-FITC/2A91 (MBL International, Woburn, MA), MPO-FITC/2C7 (AbD Serotec, Raleigh, NC), HLA-DR-APC/L243, CD68-PE/Y1-82A, MCSFR-FITC/61708 (R&D Systems, Minneapolis, MN).

Techniques: Flow Cytometry, Expressing, Isolation, In Vitro, Ex Vivo, Labeling

Journal: PLoS ONE

Article Title: Monocyte Subset Dynamics in Human Atherosclerosis Can Be Profiled with Magnetic Nano-Sensors

doi: 10.1371/journal.pone.0005663

Figure Lengend Snippet: A. Representative flow cytometry contour plots of human monocytes labeled with increasing concentrations of fluorescent superparamagnetic nano-particles CLIO-MCSFR (top row) and CLIO (bottom row). To discriminate between subsets the mean fluorescent intensity of the particle (x-axis) is plotted against expression of CD16 (y-axis). B. Fe concentrations for both nano-particles (CLIO-MCSFR and CLIO) are log-transformed (x-axes) and plotted against % of positive labeled cells (y-axes). A sigmoidal dose-response curve is generated to calculate the corresponding EC 50 (nano-particle concentration at which 50% of each monocyte subset is labeled). N = 4. C. Principle of the assay. The principle postulates that equal binding of subsets with CLIO-MCSFR will occur after 10 min at RT while incubation of subsets with CLIO at 120 min at 37°C will result in preferential uptake of the particle by CD16 lo monocytes.

Article Snippet: Cell suspensions were stained with the following antibodies (all from BD Bioscience, unless otherwise stated) at a final concentration of 1∶100: CD11b-APC-Cy7/ICRF44, CD14-PE/M5E2, CD16 PE-Cy7/3G8, CCR2-Alexa-647/48607, CX 3 CR1-FITC/2A91 (MBL International, Woburn, MA), MPO-FITC/2C7 (AbD Serotec, Raleigh, NC), HLA-DR-APC/L243, CD68-PE/Y1-82A, MCSFR-FITC/61708 (R&D Systems, Minneapolis, MN).

Techniques: Flow Cytometry, Labeling, Expressing, Transformation Assay, Generated, Concentration Assay, Binding Assay, Incubation

Journal: PLoS ONE

Article Title: Monocyte Subset Dynamics in Human Atherosclerosis Can Be Profiled with Magnetic Nano-Sensors

doi: 10.1371/journal.pone.0005663

Figure Lengend Snippet: A. Representative NIH-color coded map generated from T 2 -weighted Magnetic Resonance Imaging. Data show equal number of CD16 lo , CD16 hi monocytes and other leukocytes labeled with two CLIO-MCSFR (left panel) and CLIO (right panel) concentrations. B. T 2 measurements detected with a conventional benchtop-relaxometer. Data show equal number of CD16 lo , CD16 hi monocytes and other leukocytes labeled with two CLIO-MCSFR (left panel) and CLIO (right panel) concentrations. N = 3–5. Mean±SEM. C. T 2 changes detected with a diagnostic magnetic resonance (DMR) chip. Data show increasing number of CD16 lo , CD16 hi monocytes and other leukocytes labeled with one CLIO-MCSFR (left panel) and CLIO (right panel) concentration. N = 3. Mean±SEM. D. Two-dimensional T 2 map derived from data in C to simultaneously enumerate total monocyte numbers and subset proportions. Model combines T 2 changes for CLIO-MCSFR (x-axis) and CLIO (y-axis). Changes in predicted monocyte number are demarcated with vertical lines while the rainbow region defines monocyte subset fluctuations.

Article Snippet: Cell suspensions were stained with the following antibodies (all from BD Bioscience, unless otherwise stated) at a final concentration of 1∶100: CD11b-APC-Cy7/ICRF44, CD14-PE/M5E2, CD16 PE-Cy7/3G8, CCR2-Alexa-647/48607, CX 3 CR1-FITC/2A91 (MBL International, Woburn, MA), MPO-FITC/2C7 (AbD Serotec, Raleigh, NC), HLA-DR-APC/L243, CD68-PE/Y1-82A, MCSFR-FITC/61708 (R&D Systems, Minneapolis, MN).

Techniques: Generated, Magnetic Resonance Imaging, Labeling, Diagnostic Assay, Concentration Assay, Derivative Assay

Journal: PLoS ONE

Article Title: Monocyte Subset Dynamics in Human Atherosclerosis Can Be Profiled with Magnetic Nano-Sensors

doi: 10.1371/journal.pone.0005663

Figure Lengend Snippet: A. Representative flow cytometry dot plots of monocyte subsets from healthy volunteers and patients with documented coronary artery disease (CAD). Numbers depict percentage of subsets in both groups. Mean±SEM. B. Plots depict percentage (left plot) and absolute numbers (right plot) of CD16 lo and CD16 hi monocytes from healthy volunteers (green dots) and patients with CAD (red dots). C. Validation of the enumerative mathematical model with varying number and percentage of monocyte subsets. Data show goodness of fit of defined numbers of monocytes alone (left panel), other leukocytes (middle-left panel) and combinations of leukocytes and monocytes (right two panels). Different proportions of monocyte subsets (CD16 lo /CD16 hi (%)) are color-coded and their fit is depicted on the two-dimensional T 2 maps.

Article Snippet: Cell suspensions were stained with the following antibodies (all from BD Bioscience, unless otherwise stated) at a final concentration of 1∶100: CD11b-APC-Cy7/ICRF44, CD14-PE/M5E2, CD16 PE-Cy7/3G8, CCR2-Alexa-647/48607, CX 3 CR1-FITC/2A91 (MBL International, Woburn, MA), MPO-FITC/2C7 (AbD Serotec, Raleigh, NC), HLA-DR-APC/L243, CD68-PE/Y1-82A, MCSFR-FITC/61708 (R&D Systems, Minneapolis, MN).

Techniques: Flow Cytometry, Biomarker Discovery

Journal: Proteomes

Article Title: In-Depth Proteomic Characterization of Classical and Non-Classical Monocyte Subsets

doi: 10.3390/proteomes6010008

Figure Lengend Snippet: Immunophenotypic markers used for the identification and isolation of both classical CD14 high /CD16 − and non-classical CD16 high /CD14 −/low monocyte subsets.

Article Snippet: CD16 , PE-Cy7 , 3G8 , BD Bioscience 1.

Techniques: Isolation