flow cytometry human lyse buffer  (R&D Systems)

Journal: Journal of Innate Immunity

Article Title: Staphylococcus aureus Induces Shedding of IL-1RII in Monocytes and Neutrophils

doi: 10.1159/000443663

Figure Lengend Snippet: Shedding of IL-1RII in response to S. aureus. a, b Expression of IL-1RII on the cell surface of naive human monocytes and neutrophils as determined by flow cytometry. Boxes and whiskers depict the percentage of cells positive for IL-1RII (a) and mean fluorescence intensity of IL-1RII (b). Data are presented as the maximum and minimum values obtained from individuals donors (8 per group), and the horizontal line represents the median for each group. c-j Monocytes (c, e-g) and neutrophils (d, h-j) were stimulated for up to 2 h with S. aureus or media alone, sIL-1RII in the supernatant was quantified by ELISA (c, d) and surface expression of IL-1RII was quantified by flow cytometry (e-j). c, d Boxes and whiskers depict maximum and minimum values obtained from individual donors (10-11 per group) and the horizontal line represents the median for each group. ** p < 0.01, Wilcoxon matched-pairs signed rank test. The percentage of positive cells for IL-1RII (e, h) and mean fluorescence intensity (MFI) of IL-1RII (f, i) were determined by flow cytometry. Each pair of dots represents an individual donor. g, j The change in IL-1RII expression at the cell surface (ΔMFI) was calculated for each individual donor as the IL-1RII expression in response to S. aureus minus the IL-1RII expression in response to media. Representative histograms are shown. Open histograms represent cells stained with antibody to IL-1RII, solid histograms represent controls.

Article Snippet: Flow Cytometry Human monocytes and neutrophils were washed twice with PBS and stained with phycoerythrin-labeled anti-IL-1RII (R&D Systems).

Techniques: Expressing, Flow Cytometry, Fluorescence, Enzyme-linked Immunosorbent Assay, Staining

Journal: Journal of Innate Immunity

Article Title: Staphylococcus aureus Induces Shedding of IL-1RII in Monocytes and Neutrophils

doi: 10.1159/000443663

Figure Lengend Snippet: Peritoneal S. aureus infection. Mice were intraperitoneally inoculated with wild-type S. aureus, the SpA- mutant or PBS (control). IL-6 (a), IFN-γ (b), CXCL1 (c) and CXCL10 (d) levels in peritoneal fluid were quantified by ELISA. Each dot represents an individual mouse and horizontal lines show the median value in each group. * p < 0.05; ** p < 0.01, nonparametric Mann-Whitney test. The percentage of recruited neutrophils (e) and monocytes (f) as well as the ratio of neutrophils/monocytes (g) in the peritoneum at 4 h after inoculation was determined by flow cytometry. Boxes and whiskers depict maximum and minimum values obtained from individual mice (5 per group) and the horizontal line represents the median for each group. * p < 0.05, nonparametric Mann-Whitney test. h Bacterial counts in peritoneum at 2 and 4 h postinoculation. Each box represents an individual group of mice and horizontal lines show the median value for each group. * p < 0.05, nonparametric Mann-Whitney test.

Article Snippet: Flow Cytometry Human monocytes and neutrophils were washed twice with PBS and stained with phycoerythrin-labeled anti-IL-1RII (R&D Systems).

Techniques: Infection, Mutagenesis, Control, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Flow Cytometry