mononuclear cells mnc  (Nycomed)

Journal: Clinical and Experimental Immunology

Article Title: CD8α+ dendritic cells and immune protection from experimental allergic encephalomyelitis

doi: 10.1111/j.1365-2249.2004.02556.x

Figure Lengend Snippet: MBP87–99- and MBP68–86-induced cytokine levels. Blood was obtained on day 12 p.i., and MNC were prepared by centrifugation over Lymphoprep density gradient. IFN- γ , IL-10 and TNF- α were measured by ELISA kits. Data are presented as the mean ± s.d. of duplicate samples from three to four rats/Exp. 2 (* P

Article Snippet: Peripheral blood was obtained on day 12 p.i., and mononuclear cells (MNC) were isolated by centrifugation over Lymphoprep density gradient (Nycomed, Oslo, Norway).

Techniques: Centrifugation, Enzyme-linked Immunosorbent Assay

Journal: Clinical and Experimental Immunology

Article Title: CD8α+ dendritic cells and immune protection from experimental allergic encephalomyelitis

doi: 10.1111/j.1365-2249.2004.02556.x

Figure Lengend Snippet: Frequency of CD4 + T cell expressing IL-10. Blood was obtained on day 12 p.i. (3–4 rats/Exp. 2), and MNC were prepared by centrifugation over Lymphoprep density gradient. Immediately following separation, CD4 + T cells expressing IL-10 were stained with FITC-CD4 and PE-IL-10 antibodies, and analysed by flow cytometry. Representative patterns are shown.

Article Snippet: Peripheral blood was obtained on day 12 p.i., and mononuclear cells (MNC) were isolated by centrifugation over Lymphoprep density gradient (Nycomed, Oslo, Norway).

Techniques: Expressing, Centrifugation, Staining, Flow Cytometry, Cytometry

Journal: Clinical and Experimental Immunology

Article Title: CD8α+ dendritic cells and immune protection from experimental allergic encephalomyelitis

doi: 10.1111/j.1365-2249.2004.02556.x

Figure Lengend Snippet: Phenotypic analysis of MNC. To examine phenotypic diversity of MNC, blood was obtained on day 12 p.i. (3–4 rats/Exp. 2), and MNC were prepared by centrifugation over Lymphoprep density gradient. MNC were stained for CD4, CD8, CD28, CD40, CD80, CD86 and MHC class II, and analysed by flow cytometry. Representative patterns are shown.

Article Snippet: Peripheral blood was obtained on day 12 p.i., and mononuclear cells (MNC) were isolated by centrifugation over Lymphoprep density gradient (Nycomed, Oslo, Norway).

Techniques: Centrifugation, Staining, Flow Cytometry, Cytometry

Journal: Cell Stress & Chaperones

Article Title: Evidence of a role for both anti-Hsp70 antibody and endothelial surface membrane Hsp70 in atherosclerosis

doi: 10.1007/s12192-013-0404-4

Figure Lengend Snippet: Dose–response curves for ADCC. The procedures for OxLDL incubated and Ab treatment were the same as described in the legend for Fig. . As effectors normal rat PBMC were added to the culture instead of complement. Note increased 51Cr release

Article Snippet: To determine antibody-dependent cellular cytotoxicity (ADCC), peripheral blood mononuclear cells (PBMC) were isolated from healthy rats by density centrifugation (Lymphoprep, density 1,083; Nycomed Pharmaceuticals Oslo, Norway) as described previously (Jurgens et al. ).

Techniques: Incubation

Journal: Age

Article Title: Relationship between functional ability in older people, immune system status, and intensity of response to CMV

doi: 10.1007/s11357-011-9240-6

Figure Lengend Snippet: CD69 expression and proliferative capacity of CD4+ and CD8+ depending on the functional capacity. Whole blood from the BI groups of elders (group 0 + 1, n = 12 and group 2 + 3, n = 14) was stimulated for 18 h and expression of CD69 in CD4+ and CD8+ T cells was evaluated by flow cytometry. Proliferative capacity of CD4+ and CD8+ T cells subsets was also evaluated in the two groups (group 0 + 1, n = 19 and group 2 + 3, n = 16) by labeling the PBMC with CFSE. Cells were stained and 1 × 10 5 cells were acquired per experiment. a Representative dot plots showing the frequency of CD69 expression in CD4+ and CD8+ subset from elderly with different functional status. Cells were stimulated using anti-CD3 (10 ng/mL). Percentage of positive cells in each subpopulation in this representative experiment is expressed in the upper right corner and summarized results from all donors (median and IR) were also expressed in dot plots. b Proliferative capacity of CD4+ and CD8+ T cells subsets in response to anti-CD3. PBMC were labeled with CFSE (1.5 μM) and cultured in presence of anti-CD3 (10 ng/mL) for 5 days. Percentage of dividing CD4+ and CD8+ T cells is represented. Bars represent results from the grouped elders (mean ± SEM). c Expression of CD69 into de CD4+ and CD8+ T cell subset was analyzed in the same way as in Fig. 5a in response to a CMV supernatant (10 4 PFU/mL). d Proliferative capacity of CD4+ and CD8+ T-cell subsets in response to the CMV supernatant. Bars represent resulted from the grouped elders (mean ± SEM). The Student’s t test (when data were normally distributed) and Mann–Whitney non-parametric (when data were not normally distributed) methods were used to compare frequencies between groups. p values are depicted in the panels

Article Snippet: To analyze the proliferation status of CD4+ and CD8+ T cells, peripheral blood mononuclear cells (PBMC) were isolated by centrifugation on Ficoll-Hypaque gradients (Lymphoprep; Nycomed, Oslo, Norway).

Techniques: Expressing, Functional Assay, Flow Cytometry, Cytometry, Labeling, Staining, Cell Culture, MANN-WHITNEY

Journal: PLoS ONE

Article Title: A TNFR2-Agonist Facilitates High Purity Expansion of Human Low Purity Treg Cells

doi: 10.1371/journal.pone.0156311

Figure Lengend Snippet: TNFR2-agonist plus rapamycin expanded Treg inhibit inflammation in a humanised mouse model. (A) Schematic overview of the humanised skin inflammation mouse model used. In brief, SCID mice were transplanted with a human skin graft, 21 days after engraftment, PBS (as a control), or allogeneic human PBMC (huPBMC) only or huPBMC plus Treg of interest (at a ratio of 1:1) were injected intra peritoneally. 26 days later the animals were sacrificed to analyze the mouse spleen and human skin grafts. Rap Treg and R/T Treg refer to low purity MACS-isolated Treg expanded for 16-days in the presence of rapamycin-only or TNFR2-agonist plus rapamycin, respectively. (B) Representative photographs of spleens derived from mice infused with PBS, huPBMC only, or huPBMC plus Treg of interest, 21 days after the skin transplantation. Cumulative data showing the weight of spleens derived from N = 4 mice (right panel; n.d. = not determined). (C) Representative photographs showing histology (HE staining) of human skin grafts. Left panel: 10 x magnification. Right panel shows the epidermal thickness (μm) of human skin grafts following infusion of PBS, huPBMC, huPBMC plus Rap Treg or huPBMC plus R/T Treg. Mean ± SD, N = 6. (D) Immunohistochemistry of human CD3+ (brown) T cell infiltration in the human dermis. A representative photograph of N = 4 as presented in the cumulative data graph is shown (right panel; Mean ± SD). 20 x magnification. Wilcoxon paried t Test was used to compare the group of mice infused with huPBMC only with other groups of mice infused with huPBMC plus Treg of interest. Asterisks indicate significant differences.

Article Snippet: Isolation of Treg Peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation (Lymphoprep, Nycomed Pharma AS, Oslo, Norway) of buffy coats obtained from healthy blood donors (Sanquin Blood Bank, Region South-East, Netherlands) upon written informed consent, according to the Dutch law.

Techniques: Mouse Assay, Injection, Magnetic Cell Separation, Isolation, Derivative Assay, Transplantation Assay, Staining, Immunohistochemistry