mouse antihuman cd14 fitc conjugated  (ImmunoTools)

Journal: Frontiers in Immunology

Article Title: IL-36γ Is a Strong Inducer of IL-23 in Psoriatic Cells and Activates Angiogenesis

doi: 10.3389/fimmu.2018.00200

Figure Lengend Snippet: (A) IL-36 receptor (IL-36R) (red) on macrophages; CD14+ (green) (magnification 20×). (B) Following 48 h stimulation with IL-36γ, IL-1α, IL-17A, TNFα, and IFNγ (24 h priming), supernatant was analysed by ELISA for TNFα and IL-23. Unpaired t -test * p < 0.05, ** p < 0.01 psoriasis versus healthy (sample size: psoriasis = 9, healthy = 9, and boiled control n = 3).

Article Snippet: CD14+ purity was tested by FACs analysis with mouse antihuman CD14 FITC conjugated or mouse IgG isotype control (both 1:100; both ImmunoTools, Friesoythe, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Frontiers in Immunology

Article Title: IL-36γ Is a Strong Inducer of IL-23 in Psoriatic Cells and Activates Angiogenesis

doi: 10.3389/fimmu.2018.00200

Figure Lengend Snippet: Human umbilical vein endothelial cell monolayer was stimulated with or without TNFα 10 ng for 24 h. 1 × 10 5 monocytes were allowed to adhere to the monolayer for 30 min. Cells were visualised by immunofluorescence microscopy following CD14+ staining (B) and counted (A) (patient monocytes: psoriasis = 8; healthy = 8). Magnification 40×. Unpaired t -test * p < 0.05 and ** p < 0.01.

Article Snippet: CD14+ purity was tested by FACs analysis with mouse antihuman CD14 FITC conjugated or mouse IgG isotype control (both 1:100; both ImmunoTools, Friesoythe, Germany).

Techniques: Immunofluorescence, Microscopy, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: High-dose leptin activates human leukocytes via receptor expression on monocytes.

doi: 10.4049/jimmunol.167.8.4593

Figure Lengend Snippet: FIGURE 4. a, Dose response of luciferase activity to leptin stimulation in HEK293 cells. Cells were transiently transfected with Ob-R plasmid and a Stat3-luciferase reporter construct, and stimulated with varying doses of leptin. Luciferase activity was measured after 6-h incubation and was cor- rected for -galactosidase levels, which were used as a transfection control. The results are represented as fold induction over unstimulated cells. b, Flow cytometry characterization of the dose response for the induction of IL-6 and TNF- on CD14-gated monocytes. Whole blood was incubated with or without increasing doses of leptin or endotoxin for 6 h and cyto- kines were detected by intracellular staining in gated CD14 monocytes using two-color flow cytometry. The results are expressed as the percent- age of CD14 cells positive for IL-6 or TNF-. Leptin induced TNF- and IL-6 production in a dose-dependent manner. The response to leptin is compared with a maximal response to endotoxin. It should be noted that induction is presented in a logarithmic scale and the response to leptin is less than that seen after endotoxin.

Article Snippet: Brefeldin A (BD Biosciences), a pharmacological inhibitor of secretion, was added, and cells were incubated for 6 h followed by surface staining of monocytes using FITC-conjugated mouse antihuman CD14 (Serotec).

Techniques: Luciferase, Activity Assay, Transfection, Plasmid Preparation, Construct, Incubation, Control, Flow Cytometry, Staining, Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: High-dose leptin activates human leukocytes via receptor expression on monocytes.

doi: 10.4049/jimmunol.167.8.4593

Figure Lengend Snippet: FIGURE 5. Effects of polymyxin-B on TNF- production by endotox- in- and leptin-activated monocytes. Whole blood was treated with either endotoxin or leptin with and without polymyxin-B (1 g/ml) for 6 h and TNF- was detected by intracellular staining in gated CD14 monocytes using two-color flow cytometry. The results are expressed as the percent- age of CD14 cells positive for TNF-. Polymyxin-B blocked endotoxin induction of TNF- but had no effect on leptin activation.

Article Snippet: Brefeldin A (BD Biosciences), a pharmacological inhibitor of secretion, was added, and cells were incubated for 6 h followed by surface staining of monocytes using FITC-conjugated mouse antihuman CD14 (Serotec).

Techniques: Staining, Cytometry, Activation Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: High-dose leptin activates human leukocytes via receptor expression on monocytes.

doi: 10.4049/jimmunol.167.8.4593

Figure Lengend Snippet: FIGURE 9. Effects of ERK inhibitor and p38 inhibitor on TNF- pro- duction by either endotoxin- or leptin-activated monocytes. Whole blood was treated with and without ERK inhibitor (PD98059) and p38 inhibitor (SB203580) for 1 h before addition of either endotoxin (0.1 ng/ml) or leptin (250 ng/ml) for 6 h. TNF- was detected by intracellular staining in gated CD14 monocytes using two-color flow cytometry. The results are ex- pressed as the percentage of CD14 cells positive for TNF-. The p38 inhibitor but not the ERK inhibitor partially inhibited the induction of TNF- by both endotoxin- and leptin-activated monocytes. (, p 0.01 endotoxin only vs endotoxin plus p38 inhibitor, , p 0.01 leptin only vs leptin plus p38 inhibitor).

Article Snippet: Brefeldin A (BD Biosciences), a pharmacological inhibitor of secretion, was added, and cells were incubated for 6 h followed by surface staining of monocytes using FITC-conjugated mouse antihuman CD14 (Serotec).

Techniques: Staining, Cytometry