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Becton Dickinson blood mononuclear cells pbmcs
Frequency and molecular expression of pDC in IT, CHB and AHB groups. (a) AHB group had significantly higher <t>PBMC</t> percent in peripheral blood leukocytes, compared with CHB or IT group. (b) Frequency of pDC in PBMC decreased in AHB and CHB groups, compared with IT group. (c) The difference in CD86+ pDC frequency among three groups was not significant. (d) The counts of CD86 molecular expressed on surface of pDC in CHB group was higher, compared with AHB and IT groups. IT group: HBeAg-positive chronic hepatitis B virus infected patients in immune tolerance phase; CHB group: HBeAg-positive chronic hepatitis B infected patients; AHB group: Acute hepatitis B virus infected patients; PBMC: Peripheral blood mononuclear cells; pDC: <t>Plasmacytoid</t> dendritic cell; CD86: Cluster of differentiation antigen 86.
Blood Mononuclear Cells Pbmcs, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Plasmacytoid Dendritic Cell Function and Cytokine Network Profiles in Patients with Acute or Chronic Hepatitis B Virus Infection"

Article Title: Plasmacytoid Dendritic Cell Function and Cytokine Network Profiles in Patients with Acute or Chronic Hepatitis B Virus Infection

Journal: Chinese Medical Journal

doi: 10.4103/0366-6999.221275

Frequency and molecular expression of pDC in IT, CHB and AHB groups. (a) AHB group had significantly higher PBMC percent in peripheral blood leukocytes, compared with CHB or IT group. (b) Frequency of pDC in PBMC decreased in AHB and CHB groups, compared with IT group. (c) The difference in CD86+ pDC frequency among three groups was not significant. (d) The counts of CD86 molecular expressed on surface of pDC in CHB group was higher, compared with AHB and IT groups. IT group: HBeAg-positive chronic hepatitis B virus infected patients in immune tolerance phase; CHB group: HBeAg-positive chronic hepatitis B infected patients; AHB group: Acute hepatitis B virus infected patients; PBMC: Peripheral blood mononuclear cells; pDC: Plasmacytoid dendritic cell; CD86: Cluster of differentiation antigen 86.
Figure Legend Snippet: Frequency and molecular expression of pDC in IT, CHB and AHB groups. (a) AHB group had significantly higher PBMC percent in peripheral blood leukocytes, compared with CHB or IT group. (b) Frequency of pDC in PBMC decreased in AHB and CHB groups, compared with IT group. (c) The difference in CD86+ pDC frequency among three groups was not significant. (d) The counts of CD86 molecular expressed on surface of pDC in CHB group was higher, compared with AHB and IT groups. IT group: HBeAg-positive chronic hepatitis B virus infected patients in immune tolerance phase; CHB group: HBeAg-positive chronic hepatitis B infected patients; AHB group: Acute hepatitis B virus infected patients; PBMC: Peripheral blood mononuclear cells; pDC: Plasmacytoid dendritic cell; CD86: Cluster of differentiation antigen 86.

Techniques Used: Expressing, Infection

Frequency and molecular expression of pDC in HI and HBV infected groups. (a) The difference in percentages of PBMC in peripheral blood leukocytes between two groups was not significant. Compared with HI group, frequency of pDC in PBMC was significantly lower (b), but CD86+ pDC frequency and counts of CD86 molecular expressed on surface of pDC were higher in HBV infected group (c and d). HI: Healthy individual; HBV: Hepatitis B virus; PBMC: Peripheral blood mononuclear cells; pDC: Plasmacytoid dendritic cell; CD86: Cluster of differentiation antigen 86.
Figure Legend Snippet: Frequency and molecular expression of pDC in HI and HBV infected groups. (a) The difference in percentages of PBMC in peripheral blood leukocytes between two groups was not significant. Compared with HI group, frequency of pDC in PBMC was significantly lower (b), but CD86+ pDC frequency and counts of CD86 molecular expressed on surface of pDC were higher in HBV infected group (c and d). HI: Healthy individual; HBV: Hepatitis B virus; PBMC: Peripheral blood mononuclear cells; pDC: Plasmacytoid dendritic cell; CD86: Cluster of differentiation antigen 86.

Techniques Used: Expressing, Infection

2) Product Images from "Increased CD4+ and CD8+ effector memory T cells in patients with aplastic anemia"

Article Title: Increased CD4+ and CD8+ effector memory T cells in patients with aplastic anemia

Journal: Haematologica

doi: 10.3324/haematol.13412

Naïve and memory T-cell subsets in PBMCs and BMMNCs of patients with AA (AA, n=45) and normal controls (NC, n=30). (A) Representative flow cytometric analysis from a patient with AA and a normal control. Naïve and memory subsets were identified by the expressions of CD45RA and CCR7 after gating on CD4 + or CD8 + T cells in PBMCs and BMMNCs. (B) Percentages of naïve T cells, T CM cells and T EM cells in CD4 + T cells. (C) Percentages of naïve T cells, T CM cells, T EM cells and terminal T EM cells in CD8 + T cells. The open symbols (in B and C) represent percentages from PBMCs and solid symbols from BMMNCs. Horizontal lines indicate mean percentages.
Figure Legend Snippet: Naïve and memory T-cell subsets in PBMCs and BMMNCs of patients with AA (AA, n=45) and normal controls (NC, n=30). (A) Representative flow cytometric analysis from a patient with AA and a normal control. Naïve and memory subsets were identified by the expressions of CD45RA and CCR7 after gating on CD4 + or CD8 + T cells in PBMCs and BMMNCs. (B) Percentages of naïve T cells, T CM cells and T EM cells in CD4 + T cells. (C) Percentages of naïve T cells, T CM cells, T EM cells and terminal T EM cells in CD8 + T cells. The open symbols (in B and C) represent percentages from PBMCs and solid symbols from BMMNCs. Horizontal lines indicate mean percentages.

Techniques Used: Flow Cytometry

IFN-γ production by stimulated CCR7 + naïve/T CM cells and CCR7 − T EM cells in patients with AA (AA, n=26) and normal controls (NC, n=18). (A) Percentages of IFN-γ-producing CD4 + or CD8 + CCR7 − T EM cells and CCR7 + naïve/T CM cells among total CD4 + or CD8 + T cells from PBMCs and BMMNCs. (B) Percentages of IFN-γproducing cells in CD4 + or CD8 + CCR7 − T EM cells from PBMCs and BMMNCs. Columns represent mean ± SD values.
Figure Legend Snippet: IFN-γ production by stimulated CCR7 + naïve/T CM cells and CCR7 − T EM cells in patients with AA (AA, n=26) and normal controls (NC, n=18). (A) Percentages of IFN-γ-producing CD4 + or CD8 + CCR7 − T EM cells and CCR7 + naïve/T CM cells among total CD4 + or CD8 + T cells from PBMCs and BMMNCs. (B) Percentages of IFN-γproducing cells in CD4 + or CD8 + CCR7 − T EM cells from PBMCs and BMMNCs. Columns represent mean ± SD values.

Techniques Used:

3) Product Images from "Activated CD8 T cells acquire NK1.1 expression and preferentially locate in the liver in mice after allogeneic hematopoietic cell transplantation"

Article Title: Activated CD8 T cells acquire NK1.1 expression and preferentially locate in the liver in mice after allogeneic hematopoietic cell transplantation

Journal: Immunology letters

doi: 10.1016/j.imlet.2012.12.009

NK1.1 + CD8 + T cells with an effector/memory phenotype, which were derived from donor splenocytes, preferentially located in the liver of allo-HCT recipients. Sublethally-irradiated B6D2F1 mice were administered 5×10 6 BMCs and 2×10 7 splenocytes from allogeneic B6 donors. Ficolled liver cells, splenocytes, PBMCs, BMCs, and thymocytes were prepared 56 days after HCT for analyses. A) Representative FACS profiles showing staining of ficolled liver cells with anti-mouse TCRαβ and NK1.1 (left); NK1.1 + TCRαβ + cells (indicated as arrow pointed cells) were then analyzed for CD4 and CD8 expression (middle); and CD8 + NK1.1 + TCRαβ + cells (indicated as arrow pointed cells) were further analyzed for CD44 and CD62L expression (right). B) Percentages (mean±SEMs) of NK1.1 + CD4 + , NK1.1 + CD8 + and NK1.1 + CD4 − CD8 − cells in the liver (ficolled liver cells) of allo-HCT recipients. * p
Figure Legend Snippet: NK1.1 + CD8 + T cells with an effector/memory phenotype, which were derived from donor splenocytes, preferentially located in the liver of allo-HCT recipients. Sublethally-irradiated B6D2F1 mice were administered 5×10 6 BMCs and 2×10 7 splenocytes from allogeneic B6 donors. Ficolled liver cells, splenocytes, PBMCs, BMCs, and thymocytes were prepared 56 days after HCT for analyses. A) Representative FACS profiles showing staining of ficolled liver cells with anti-mouse TCRαβ and NK1.1 (left); NK1.1 + TCRαβ + cells (indicated as arrow pointed cells) were then analyzed for CD4 and CD8 expression (middle); and CD8 + NK1.1 + TCRαβ + cells (indicated as arrow pointed cells) were further analyzed for CD44 and CD62L expression (right). B) Percentages (mean±SEMs) of NK1.1 + CD4 + , NK1.1 + CD8 + and NK1.1 + CD4 − CD8 − cells in the liver (ficolled liver cells) of allo-HCT recipients. * p

Techniques Used: Derivative Assay, Irradiation, Mouse Assay, FACS, Staining, Expressing

4) Product Images from "An exploratory double-blind, randomized clinical trial with selisistat, a SirT1 inhibitor, in patients with Huntington’s disease"

Article Title: An exploratory double-blind, randomized clinical trial with selisistat, a SirT1 inhibitor, in patients with Huntington’s disease

Journal: British Journal of Clinical Pharmacology

doi: 10.1111/bcp.12512

(A) Model predicted values and 95% confidence intervals of estimated log soluble Htt levels in peripheral blood mononuclear cells as a proportion of total protein by group at each time point. No statistically significant differences were observed between
Figure Legend Snippet: (A) Model predicted values and 95% confidence intervals of estimated log soluble Htt levels in peripheral blood mononuclear cells as a proportion of total protein by group at each time point. No statistically significant differences were observed between

Techniques Used:

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Article Snippet: Isolation of peripheral blood mononuclear cells (PBMCs) from human blood Human PBMCs were isolated from whole blood with the help of BD Vacutainer® CPTTM and cultured in RPMI medium.

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Article Snippet: Blood was drawn from a cubital vein and collected into sterile EDTA tubes for isolation of peripheral blood mononuclear cells (PBMCs) or from serum tubes (BD Biosciences, Franklin Lakes, NJ, USA).

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Article Snippet: In order to assess messenger RNA (mRNA) levels of the glucocorticoid receptor (GR) gene NR3C1 , total RNA was extracted from peripheral blood mononuclear cells (PBMCs) following the manufacturer protocol (Becton Dickinson and Co., East Rutherford, NJ).

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Article Snippet: For the isolation of peripheral blood mononuclear cells (PBMCs), blood samples were collected in Vacutainer Plus (Plastic) Sterile Evacuated K2 EDTA spray dried Blood Collection Tubes (BD, Plymouth, UK).

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Article Snippet: For the in vitro stimulation of peripheral blood mononuclear cells (PBMCs), whole blood was collected 24 h after NPT (before and after each product consumption period), in sodium citrate tubes (Vacutainer® CPT™, BD, Basel, Switzerland), and PBMCs purified by centrifugation over a density gradient.

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Article Snippet: After washing, 106 peripheral blood mononuclear cells (PBMCs) were added to each well along with anti-human CD107a allophycocyanin-H7 Ab (clone H4A3; BD Biosciences), 5 μg/ml brefeldin A (Sigma-Aldrich), and 5 μg/ml monensin (Golgi Stop; BD Biosciences) for 5 hours at 37°C with 5% CO2 .

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Article Snippet: Isolation of peripheral blood mononuclear cells (PBMCs) 5 ml blood sample was collected in ethylenediaminetetra acetic acid (EDTA) vials (BD Biosciences, USA) from each study subject through an antecubital vein (phlebotomy) after an overnight fast.

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Article Snippet: Whole blood was shipped overnight express at room temperature to the University of Colorado‐Anschutz Medical Campus for Ficoll‐gradient purification of peripheral blood mononuclear cells (PBMCs) and dextran‐gradient purification of neutrophils according to standard protocols (BD Biosciences, San Jose, CA).

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Article Snippet: Peripheral blood processing For isolation of peripheral blood mononuclear cells (PBMCs), whole blood from PUUV-infected patients and UC was collected in CPT tubes (BD) and centrifuged according to manufacturer’s instructions.

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Article Snippet: Single cell suspensions, including peripheral blood mononuclear cells (PBMCs), splenocytes, BMCs, thymocytes, and ficolled liver cells, were stained by various combinations of the following fluorescence-conjugated anti-mouse mAbs: CD4, CD8, CD44, CD62L, TCRαβ, CD49b, Ly-49c, Ly-49D, Ly-49G2 (Becton Dickinson, San Jose, CA), and CD1d tetramer loaded with an analog of the αGalCer ligand, PBS57 (National Institute of Allergy and Infectious Diseases Tetramer Facility, Germantown, MD).

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Article Snippet: Freshly isolated peripheral blood mononuclear cells (PBMCs) and bone marrow mononuclear cells (BMMNCs) were stained with FITC-conjugated anti-CD45RA, PE-conjugated anti-CCR7, and PE-Cy5-conjugated anti-CD4 or PE-Cy5-conjugated anti-CD8 (BD Biosciences, San Diego, CA, USA) to measure percentages of naïve T cells, TCM cells and TEM cells in CD4+ and CD8+ T cells in both PBMCs and BMMNCs.

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Article Snippet: Briefly, 1 × 106 peripheral blood mononuclear cells (PBMCs) were stained with the appropriate antibodies for 20 min at 4 °C in the dark, and then washed and acquired using FACSVerse™ cytometer (BD Biosciences).

Article Title:
Article Snippet: Briefly, 2 × 106 peripheral blood mononuclear cells (PBMCs) were incubated in 1 ml of RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin that contained monensin (0.7 μg/ml; BD Biosciences) and brefeldin A (10 μg/ml; Sigma-Aldrich) in the absence or presence of CD107a MAbs and peptides (15mers overlapping by 11 residues) corresponding to full-length SIVmac239 Gag (2 μg/ml for each peptide at 5 μl/ml; National Institutes of Health AIDS Research and Reference Reagent Program) for 6 h. After washing, cells were surface stained for CD4, CD8, CD28, and CD95; aqua amine reactive dye was used to exclude dead cells from the analysis.

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Article Snippet: Then we separated the peripheral blood mononuclear cells (PBMCs) by centrifugation at 200g for 15 min with Ficoll-Paque (BD) from the whole blood.

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Article Snippet: Flow cytometry (FCM) immunophenotypic analysis was performed on peripheral blood mononuclear cells (PBMCs) using a FACSCalibur flow cytometer (Becton-Dickinson, San Jose, CA).

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Article Snippet: One hundred microliters of peripheral blood mononuclear cells (PBMCs) was incubated with CD4-PerCP-Cy5.5, CD3-Alexa700, CD14-PerCP-Cy5.5, and CD20-PerCP-Cy5.5, HLA-DR− APC-Cy7) (Becton-Dickinson); CD11c-APC (Pharmingen, San Jose, CA); and PE-conjugated antibody to CD123.

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Article Snippet: Cryopreserved peripheral blood mononuclear cells (PBMCs) were thawed and resuspended in RPMI 1640 plus 10% human Ab serum with the addition of anti-CD28 and -CD49d monoclonal antibodies (mAbs) (1 μg/ml; BD Biosciences, San Jose, CA).

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Article Snippet: Isolation of peripheral blood mononuclear cells (PBMCs) 5 mL of blood was obtained by cardiac puncture under anesthesia with isoflurane and collected in BD Vacutainer® blood collection tubes containing EDTA (BD, Franklin Lakes, NJ, USA).

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Article Snippet: Blood samples for determination of intracellular pharmacokinetic measurements in peripheral blood mononuclear cells (PBMCs) were collected in two 8-ml cell preparation tubes (CPT; Becton Dickinson Vacutainer) per time point.

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Article Snippet: Blood samples for measurement of soluble HTT were collected in Becton Dickinson CPT™ tubes for preparation of peripheral blood mononuclear cells (PBMCs) and plasma for immune markers was collected in Becton Dickinson K2 EDTA Vacutainer® tubes.

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Article Snippet: Circulating peripheral blood mononuclear cells (PBMCs) isolated by density gradient centrifugation and infiltrating lymphocytes dissociated from fresh tumor or non-malignant tonsil digests were stained with the following mAbs: anti-human CD3-PERCP, CD8-APC, CD4-FITC (BD Biosciences, Mountain View, CA), and anti-human PD-1 (MDX-1106, fully human IgG4, 10 ug/ml, BMS, New York, NY) or an isotype control, anti-diphtheria toxin human IgG4 (BMS).

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Article Snippet: All analyzes were accredited after International and European standard NS-EN ISO 15189 Expression of lipid-related genes in peripheral blood mononuclear cells (PBMCs) After blood collection, PBMCs were isolated by using the BD Vacutainer Cell Preparation tubes with sodium heparin according to the manufacturer (Becton Dickinson, San Jose, CA) and stored as pellets at -80o C until further mRNA isolation.

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Article Snippet: CellROX® Orange Reagent (Life Technologies, Carlsbad, CA, USA) was used for measuring intracellular reactive oxygen species (ROS) production in peripheral blood mononuclear cells (PBMCs) using a FACScanto II analyzer (Becton–Dickinson, San Diego, CA, USA) and Flow-Jo v10 software (Tree Star Inc., Ashland, OR, USA); intracellular ROS production (CellROX) was measured as percent positive cells (%) and mean fluorescence intensity (MFI).

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Article Snippet: Blood mononuclear cells (PBMCs) or isolated Tregs were analyzed for surface expression or intracellular markers using the following anti-human antibodies: PE-Cy7-labeled anti-CD4 (clone:SK3, BD Biosciences), Percp-Cy5.5-labeled anti-CD25 (clone: M-A251, BD Biosciences), APC-Cy7-labeled anti-CD39 (clone: A1, BD Biosciences), BV421-labeled anti-CTLA4 (clone: BNI3, BioLegend, San Diego, USA), PE-labeled anti-PD-L1 (clone: 29E2A3, BioLegend), Alexa Fluor 647-labeled anti-FOXP3 (clone: 259D/C7, BD Biosciences), Alexa Fluor 488-labeled anti-Helios (clone: 22F6, BD Biosciences), PE-labeled IL-10(clone: JES3-19F1, BioLegend) and PE-labeled anti-GITRL(clone: REA841, Miltenyi Biotec, Bergisch Gladbach, Germany).

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Article Snippet: To isolate the peripheral blood mononuclear cells (PBMCs), blood was collected through the brachial wing vein using a 5 mL syringe equipped with a 26-gauge (0.45 mm diameter) needle and stored in BD Vacutainer tubes coated with sodium heparin (BD Biosciences, Franklin Lakes, NJ, USA).

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Article Snippet: The frequencies of CD4+ and CD8+ T cells expressing specific markers (IL-2, TNF-α, IFN-γ, or CD40L [BD Biosciences]) upon in vitro stimulation of peripheral blood mononuclear cells (PBMCs) with pools of peptides covering the sequences of the p17, p24, RT, and Nef antigens were determined by flow cytometry (LSRII cytometer; Becton Dickinson) using intracellular cytokine staining (ICS), as previously described ( ).

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Article Snippet: 2.3 Expression analysis RNA was extracted from peripheral blood mononuclear cells (PBMCs) from fresh blood by centrifugation in CPT tubes (Becton Dickinson), using the Qiamp RNA Blood Mini kit (Qiagen).

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Article Snippet: The peripheral DC subset frequency was analyzed using protocols described previously by our team and with minor modifications., In brief, freshly isolated peripheral blood mononuclear cells (PBMCs) were incubated with anti-lineage-1-FITC, anti-HLA-DR-PerCP and anti-CD11c-APC (BD Pharmingen, San Jose, CA, USA) or anti-CD123-APC (Miltenyi Biotec, Bergish Gladbach, Germany).

Article Title:
Article Snippet: Frequency and molecular expression of plasmacytoid dendritic cell In this study, the peripheral blood mononuclear cells (PBMCs) percentage of peripheral blood leukocytes, frequency of pDC in PBMC, frequency of cluster of differentiation antigen 86 (CD86) + pDC, and the counts of CD86 molecular expressed on surface of pDC were measured by four-color flow cytometry (FACS Caliburflow Cytometer; Becton-Dickinson, USA); and the steps to measure the expression of pDCs were as follows: the 100 μl of whole peripheral blood samples were incubated with monoclonal antibodies (mAbs) of Lin1-fluorescein isothiocyanate, human leukocyte antigen DR (HLA-DR)-peridinin chlorophyll protein, CD123-human antigen presenting cells, and CD86 - phycoerythrin (PE; all provided by BD Biosciences, Cowley, UK) inappropriate tubes at room temperature in the dark for 20 min, then added 2 ml of fluorescence activating cell sorter (FACS) Lysing solution and incubated at room temperature in the dark for 5 min, centrifuged at 300 ×g for 5 min, aspirated the supernatant, then added 2 ml of phosphate buffer saline (PBS) and vortex gently, centrifuged at 300 ×g for 5 min and aspirated the supernatant again, vortex gently and re-suspended with 200 μl PBS; finally analyzed using the FACS flow cytometer. pDCs were identified as mononuclear cells that the Lin1-(CD3-CD14-CD16-CD19-CD20-)/HLA-DR+/CD123+, the activation/maturation state of pDC was assessed by the co-stimulatory molecules CD86 in Lin1-CD123+ HLA-DR+cells.

Article Title:
Article Snippet: Flow Cytometry and Cell Sorting T cell subsets of interest were flow-sorted from freshly isolated peripheral blood mononuclear cells (PBMCs) at > 98% purity using a FACSVantage SE, a FACSAria, or a Special Order Research Product FACSAria II (all from BD Biosciences).

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    Becton Dickinson pbmcs
    Flow <t>cytometric</t> analyses of interferon gamma (IFN-γ) expression on NK cells and NK cell subsets in <t>PBMCs</t> isolated from normal-weight (nw) and obese (ob) individuals. a , b Exemplary FACS plots of IFN-γ expression in CD56 bright and CD56 dim NK cells of a normal-weight and an obese individual. c Frequency of IFN-γ-expressing NK cells in PBMCs. d , e IFN-γ expression in CD56 bright ( d ) and CD56 dim ( e ) NK cells. Data are expressed as mean ± SEM. * P
    Pbmcs, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 1 article reviews
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    Becton Dickinson peripheral blood mononuclear cells pbmcs
    Irradiation-induced angiogenesis is dictated by MMP-9–mediated factor release from mast and stroma cells and incorporation of BM-derived cells. (A–C) HL ischemia was induced in Sl/Sl d and WBB6F1 +/+ mice, followed by 2 Gy IR or no IR ( n = 7). <t>PBMCs</t> were analyzed for the presence of CFU-Cs (A) and CFU-ECs (B). (C) Plasma VEGF was determined ( n = 7/group). (D) HL ischemia was induced in MMP-9 +/+ mice followed by 2 Gy IR. Mice were subdivided into groups receiving neutralizing <t>mAbs</t> against VEGFR-1, VEGFR-2 or a combination of both mAbs. (a) Blood flow was determined. Insert: Percentage of mice showing limb rescue after ligation. (b) Macroscopic pictures taken of ischemic and contralateral limb 7 d after ligation (*P
    Peripheral Blood Mononuclear Cells Pbmcs, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    CXCR3 and CCR6 expression in T cells of major depressive disorder (MDD) patients and non-depressed controls. (A) CXCR3-expressing T cells were identified by flow cytometric analysis of peripheral blood mononuclear cells from MDD patients and matched non-depressed controls (CTR). Displayed values are frequencies of CXCR3 + T cells expressed as a percentage of live CD3 + lymphocytes from a representative case–control pair. (B) Percentages of CXCR3-expressing total T cells, <t>CD4</t> + , and CD8 + T cells were quantified in our cohort ( n = 40). (C,D) Similar analyses were conducted for the surface expression of CCR6 on total T cells as well as on the CD4 + and CD8 + T cell subsets. (E) The CXCR3 ligands CXCL10 and CXCL11 were quantified in sera of MDD patients and matched controls using a cytometric bead array ( n = 38). (F) Surface CD3 MFI levels were measured by flow cytometric analysis of CD4 + and CD8 + T cells from MDD patients and matched controls ( n = 40). All graphs depict medians with interquartile ranges. For all comparisons, the Wilcoxon signed-rank test was used. SSC-A, side scatter-area; CXCR3, CXC-chemokine receptor type 3; CCR6, CC-chemokine receptor type 6; MFI, median fluorescence intensity.
    Pbmc Aliquots, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Signal scatter plots comparing the intensities of gene expressions on microarray chips within <t>Ficoll</t> and CPT-derived <t>PBMC</t> and their immune cell subsets for each healthy donor tested. The expression signals of Ficoll-derived cells were plotted against the expression signals of CPT-derived cells for each cell type and donor. A line of perfect correlation is indicated in all plots. Pearson’s correlation coefficients between each pair of samples are shown at the bottom right of each plot. The data showed that the pattern of gene expression was very similar between Ficoll and CPT-derived cell types, as indicated by the high correlation coefficients
    Pbmc, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Flow cytometric analyses of interferon gamma (IFN-γ) expression on NK cells and NK cell subsets in PBMCs isolated from normal-weight (nw) and obese (ob) individuals. a , b Exemplary FACS plots of IFN-γ expression in CD56 bright and CD56 dim NK cells of a normal-weight and an obese individual. c Frequency of IFN-γ-expressing NK cells in PBMCs. d , e IFN-γ expression in CD56 bright ( d ) and CD56 dim ( e ) NK cells. Data are expressed as mean ± SEM. * P

    Journal: Immunologic Research

    Article Title: Impaired natural killer cell subset phenotypes in human obesity

    doi: 10.1007/s12026-018-8989-4

    Figure Lengend Snippet: Flow cytometric analyses of interferon gamma (IFN-γ) expression on NK cells and NK cell subsets in PBMCs isolated from normal-weight (nw) and obese (ob) individuals. a , b Exemplary FACS plots of IFN-γ expression in CD56 bright and CD56 dim NK cells of a normal-weight and an obese individual. c Frequency of IFN-γ-expressing NK cells in PBMCs. d , e IFN-γ expression in CD56 bright ( d ) and CD56 dim ( e ) NK cells. Data are expressed as mean ± SEM. * P

    Article Snippet: For flow cytometric analyses, PBMCs were stained with the following antibodies (all from BD Biosciences, San Diego, USA, unless otherwise indicated): CD3 conjugated with phycoerythrin (PE)-Cy 7, CD4 conjugated with allophycocyanin (APC), CD 8 conjugated with PE, CD14 conjugated with fluorescein isothiocyanate (FITC), CD20 conjugated with APC-H7, CD56 conjugated with APC, CD253 (TRAIL; tumor necrosis factor-related apoptosis-inducing ligand) conjugated with PE, Ob-R (leptin receptor) conjugated with fluorescein (R & D Systems, Minneapolis, MN, USA), CD314 (NKG2D) conjugated with PE-CF594, CD25 conjugated with PE CF594, CD69 conjugated with FITC, and CD107a conjugated with PE.

    Techniques: Flow Cytometry, Expressing, Isolation, FACS

    Flow cytometric analyses of NK cells and NK cell subsets in PBMCs (peripheral blood mononuclear cells) isolated from normal-weight (nw) and obese (ob) individuals. a , b Exemplary FACS plots of CD56 bright and CD56 dim NK cells of a normal-weight and an obese subject. c Frequency of NK cells in PBMCs. d , e Expression of CD56 bright ( d ) and CD56 dim ( e ) NK cells. Data are expressed as mean ± SEM. * P

    Journal: Immunologic Research

    Article Title: Impaired natural killer cell subset phenotypes in human obesity

    doi: 10.1007/s12026-018-8989-4

    Figure Lengend Snippet: Flow cytometric analyses of NK cells and NK cell subsets in PBMCs (peripheral blood mononuclear cells) isolated from normal-weight (nw) and obese (ob) individuals. a , b Exemplary FACS plots of CD56 bright and CD56 dim NK cells of a normal-weight and an obese subject. c Frequency of NK cells in PBMCs. d , e Expression of CD56 bright ( d ) and CD56 dim ( e ) NK cells. Data are expressed as mean ± SEM. * P

    Article Snippet: For flow cytometric analyses, PBMCs were stained with the following antibodies (all from BD Biosciences, San Diego, USA, unless otherwise indicated): CD3 conjugated with phycoerythrin (PE)-Cy 7, CD4 conjugated with allophycocyanin (APC), CD 8 conjugated with PE, CD14 conjugated with fluorescein isothiocyanate (FITC), CD20 conjugated with APC-H7, CD56 conjugated with APC, CD253 (TRAIL; tumor necrosis factor-related apoptosis-inducing ligand) conjugated with PE, Ob-R (leptin receptor) conjugated with fluorescein (R & D Systems, Minneapolis, MN, USA), CD314 (NKG2D) conjugated with PE-CF594, CD25 conjugated with PE CF594, CD69 conjugated with FITC, and CD107a conjugated with PE.

    Techniques: Flow Cytometry, Isolation, FACS, Expressing

    Flow cytometric analyses of NKG2D receptor expression on NK cells and NK cell subsets in PBMCs isolated from normal-weight (nw) and obese (ob) individuals. a , b Exemplary FACS plots of NKG2D expression in CD56 bright and CD56 dim NK cells of a normal-weight and an obese subject. c Frequency of NKG2D-positive NK cells in PBMCs. d , e Expression of NKG2D in CD56 bright ( d ) and CD56 dim ( e ) NK cells. Data are expressed as mean ± SEM. * P

    Journal: Immunologic Research

    Article Title: Impaired natural killer cell subset phenotypes in human obesity

    doi: 10.1007/s12026-018-8989-4

    Figure Lengend Snippet: Flow cytometric analyses of NKG2D receptor expression on NK cells and NK cell subsets in PBMCs isolated from normal-weight (nw) and obese (ob) individuals. a , b Exemplary FACS plots of NKG2D expression in CD56 bright and CD56 dim NK cells of a normal-weight and an obese subject. c Frequency of NKG2D-positive NK cells in PBMCs. d , e Expression of NKG2D in CD56 bright ( d ) and CD56 dim ( e ) NK cells. Data are expressed as mean ± SEM. * P

    Article Snippet: For flow cytometric analyses, PBMCs were stained with the following antibodies (all from BD Biosciences, San Diego, USA, unless otherwise indicated): CD3 conjugated with phycoerythrin (PE)-Cy 7, CD4 conjugated with allophycocyanin (APC), CD 8 conjugated with PE, CD14 conjugated with fluorescein isothiocyanate (FITC), CD20 conjugated with APC-H7, CD56 conjugated with APC, CD253 (TRAIL; tumor necrosis factor-related apoptosis-inducing ligand) conjugated with PE, Ob-R (leptin receptor) conjugated with fluorescein (R & D Systems, Minneapolis, MN, USA), CD314 (NKG2D) conjugated with PE-CF594, CD25 conjugated with PE CF594, CD69 conjugated with FITC, and CD107a conjugated with PE.

    Techniques: Flow Cytometry, Expressing, Isolation, FACS

    Flow cytometric analyses of leptin receptor (Ob-R) expression in NK cells and NK cell subsets in PBMCs isolated from normal-weight (nw) and obese (ob) individuals. a Frequency of Ob-R expressing NK cells in PBMCs. b , c Expression of Ob-R in CD56 bright ( b ) and CD56 dim ( c ) NK cells. Data are expressed as mean ± SEM

    Journal: Immunologic Research

    Article Title: Impaired natural killer cell subset phenotypes in human obesity

    doi: 10.1007/s12026-018-8989-4

    Figure Lengend Snippet: Flow cytometric analyses of leptin receptor (Ob-R) expression in NK cells and NK cell subsets in PBMCs isolated from normal-weight (nw) and obese (ob) individuals. a Frequency of Ob-R expressing NK cells in PBMCs. b , c Expression of Ob-R in CD56 bright ( b ) and CD56 dim ( c ) NK cells. Data are expressed as mean ± SEM

    Article Snippet: For flow cytometric analyses, PBMCs were stained with the following antibodies (all from BD Biosciences, San Diego, USA, unless otherwise indicated): CD3 conjugated with phycoerythrin (PE)-Cy 7, CD4 conjugated with allophycocyanin (APC), CD 8 conjugated with PE, CD14 conjugated with fluorescein isothiocyanate (FITC), CD20 conjugated with APC-H7, CD56 conjugated with APC, CD253 (TRAIL; tumor necrosis factor-related apoptosis-inducing ligand) conjugated with PE, Ob-R (leptin receptor) conjugated with fluorescein (R & D Systems, Minneapolis, MN, USA), CD314 (NKG2D) conjugated with PE-CF594, CD25 conjugated with PE CF594, CD69 conjugated with FITC, and CD107a conjugated with PE.

    Techniques: Flow Cytometry, Expressing, Isolation

    Flow cytometric analyses of CD69 expression on NK cells and NK cell subsets in PBMCs isolated from normal-weight (nw) and obese (ob) individuals. a , b Exemplary FACS plots of CD69 expression in CD56 bright and CD56 dim NK cells of a normal-weight and an obese subject. c Frequency of CD69-positive NK cells in PBMCs. d , e Expression of CD69 in CD56 bright ( d ) and CD56 dim ( e ) NK cells. Data are expressed as mean ± SEM. * P

    Journal: Immunologic Research

    Article Title: Impaired natural killer cell subset phenotypes in human obesity

    doi: 10.1007/s12026-018-8989-4

    Figure Lengend Snippet: Flow cytometric analyses of CD69 expression on NK cells and NK cell subsets in PBMCs isolated from normal-weight (nw) and obese (ob) individuals. a , b Exemplary FACS plots of CD69 expression in CD56 bright and CD56 dim NK cells of a normal-weight and an obese subject. c Frequency of CD69-positive NK cells in PBMCs. d , e Expression of CD69 in CD56 bright ( d ) and CD56 dim ( e ) NK cells. Data are expressed as mean ± SEM. * P

    Article Snippet: For flow cytometric analyses, PBMCs were stained with the following antibodies (all from BD Biosciences, San Diego, USA, unless otherwise indicated): CD3 conjugated with phycoerythrin (PE)-Cy 7, CD4 conjugated with allophycocyanin (APC), CD 8 conjugated with PE, CD14 conjugated with fluorescein isothiocyanate (FITC), CD20 conjugated with APC-H7, CD56 conjugated with APC, CD253 (TRAIL; tumor necrosis factor-related apoptosis-inducing ligand) conjugated with PE, Ob-R (leptin receptor) conjugated with fluorescein (R & D Systems, Minneapolis, MN, USA), CD314 (NKG2D) conjugated with PE-CF594, CD25 conjugated with PE CF594, CD69 conjugated with FITC, and CD107a conjugated with PE.

    Techniques: Flow Cytometry, Expressing, Isolation, FACS

    Irradiation-induced angiogenesis is dictated by MMP-9–mediated factor release from mast and stroma cells and incorporation of BM-derived cells. (A–C) HL ischemia was induced in Sl/Sl d and WBB6F1 +/+ mice, followed by 2 Gy IR or no IR ( n = 7). PBMCs were analyzed for the presence of CFU-Cs (A) and CFU-ECs (B). (C) Plasma VEGF was determined ( n = 7/group). (D) HL ischemia was induced in MMP-9 +/+ mice followed by 2 Gy IR. Mice were subdivided into groups receiving neutralizing mAbs against VEGFR-1, VEGFR-2 or a combination of both mAbs. (a) Blood flow was determined. Insert: Percentage of mice showing limb rescue after ligation. (b) Macroscopic pictures taken of ischemic and contralateral limb 7 d after ligation (*P

    Journal: The Journal of Experimental Medicine

    Article Title: Low-dose irradiation promotes tissue revascularization through VEGF release from mast cells and MMP-9-mediated progenitor cell mobilization

    doi: 10.1084/jem.20050959

    Figure Lengend Snippet: Irradiation-induced angiogenesis is dictated by MMP-9–mediated factor release from mast and stroma cells and incorporation of BM-derived cells. (A–C) HL ischemia was induced in Sl/Sl d and WBB6F1 +/+ mice, followed by 2 Gy IR or no IR ( n = 7). PBMCs were analyzed for the presence of CFU-Cs (A) and CFU-ECs (B). (C) Plasma VEGF was determined ( n = 7/group). (D) HL ischemia was induced in MMP-9 +/+ mice followed by 2 Gy IR. Mice were subdivided into groups receiving neutralizing mAbs against VEGFR-1, VEGFR-2 or a combination of both mAbs. (a) Blood flow was determined. Insert: Percentage of mice showing limb rescue after ligation. (b) Macroscopic pictures taken of ischemic and contralateral limb 7 d after ligation (*P

    Article Snippet: To understand which cell types increase after IR, we analyzed peripheral blood mononuclear cells (PBMCs) via FACS (Becton Dickinson) using mAbs against c-kit and VEGFR-2 after IR.

    Techniques: Irradiation, Derivative Assay, Mouse Assay, Flow Cytometry, Ligation

    CXCR3 and CCR6 expression in T cells of major depressive disorder (MDD) patients and non-depressed controls. (A) CXCR3-expressing T cells were identified by flow cytometric analysis of peripheral blood mononuclear cells from MDD patients and matched non-depressed controls (CTR). Displayed values are frequencies of CXCR3 + T cells expressed as a percentage of live CD3 + lymphocytes from a representative case–control pair. (B) Percentages of CXCR3-expressing total T cells, CD4 + , and CD8 + T cells were quantified in our cohort ( n = 40). (C,D) Similar analyses were conducted for the surface expression of CCR6 on total T cells as well as on the CD4 + and CD8 + T cell subsets. (E) The CXCR3 ligands CXCL10 and CXCL11 were quantified in sera of MDD patients and matched controls using a cytometric bead array ( n = 38). (F) Surface CD3 MFI levels were measured by flow cytometric analysis of CD4 + and CD8 + T cells from MDD patients and matched controls ( n = 40). All graphs depict medians with interquartile ranges. For all comparisons, the Wilcoxon signed-rank test was used. SSC-A, side scatter-area; CXCR3, CXC-chemokine receptor type 3; CCR6, CC-chemokine receptor type 6; MFI, median fluorescence intensity.

    Journal: Frontiers in Immunology

    Article Title: T Cell Phenotype and T Cell Receptor Repertoire in Patients with Major Depressive Disorder

    doi: 10.3389/fimmu.2018.00291

    Figure Lengend Snippet: CXCR3 and CCR6 expression in T cells of major depressive disorder (MDD) patients and non-depressed controls. (A) CXCR3-expressing T cells were identified by flow cytometric analysis of peripheral blood mononuclear cells from MDD patients and matched non-depressed controls (CTR). Displayed values are frequencies of CXCR3 + T cells expressed as a percentage of live CD3 + lymphocytes from a representative case–control pair. (B) Percentages of CXCR3-expressing total T cells, CD4 + , and CD8 + T cells were quantified in our cohort ( n = 40). (C,D) Similar analyses were conducted for the surface expression of CCR6 on total T cells as well as on the CD4 + and CD8 + T cell subsets. (E) The CXCR3 ligands CXCL10 and CXCL11 were quantified in sera of MDD patients and matched controls using a cytometric bead array ( n = 38). (F) Surface CD3 MFI levels were measured by flow cytometric analysis of CD4 + and CD8 + T cells from MDD patients and matched controls ( n = 40). All graphs depict medians with interquartile ranges. For all comparisons, the Wilcoxon signed-rank test was used. SSC-A, side scatter-area; CXCR3, CXC-chemokine receptor type 3; CCR6, CC-chemokine receptor type 6; MFI, median fluorescence intensity.

    Article Snippet: Cell Purification For qRT-PCR and TCR sequencing analyses, CD4+ T cells were purified using magnetic beads (negative selection by BD IMag Human CD4 T Lymphocyte Enrichment Set, BD Biosciences) from PBMC aliquots thawed in cell separation buffer (1% human serum, 2 mM EDTA in PBS).

    Techniques: Expressing, Flow Cytometry, Fluorescence

    Surface and intracellular staining of CXCR3. (A) Percentages of CXCR3- and CCR6-expressing CD3 − lymphocytes (non-T cells) were quantified in major depressive disorder (MDD) patients and matched non-depressed controls (CTR) ( n = 40). (B) A representative plot shows fluorescence intensity of CXCR3 expression in intact (surface CXCR3; light gray-shaded curve) relative to fixed and permeabilized T cells (total cellular CXCR3; dark gray-shaded curve). Isotype-matched negative controls were used at the same concentration before fixation (black-dashed curve) and after fixation-permeabilization (gray-dashed curve) and showed no positive staining for CXCR3. (C) Total cellular CXCR3 MFI levels were measured by flow cytometric analysis of fixed and permeabilized peripheral blood mononuclear cells (PBMCs) from MDD patients and matched controls ( n = 36). Stained PBMCs were gated on live CD3 + lymphocytes (T cells), CD4 + and CD8 + T cell subsets as well as CD3 − lymphocytes (non-T cells). Graphs depict medians with interquartile ranges. For all comparisons, the Wilcoxon signed-rank test was used. CXCR3: CXC-chemokine receptor type 3; CCR6: CC-chemokine receptor type 6; MFI: median fluorescence intensity.

    Journal: Frontiers in Immunology

    Article Title: T Cell Phenotype and T Cell Receptor Repertoire in Patients with Major Depressive Disorder

    doi: 10.3389/fimmu.2018.00291

    Figure Lengend Snippet: Surface and intracellular staining of CXCR3. (A) Percentages of CXCR3- and CCR6-expressing CD3 − lymphocytes (non-T cells) were quantified in major depressive disorder (MDD) patients and matched non-depressed controls (CTR) ( n = 40). (B) A representative plot shows fluorescence intensity of CXCR3 expression in intact (surface CXCR3; light gray-shaded curve) relative to fixed and permeabilized T cells (total cellular CXCR3; dark gray-shaded curve). Isotype-matched negative controls were used at the same concentration before fixation (black-dashed curve) and after fixation-permeabilization (gray-dashed curve) and showed no positive staining for CXCR3. (C) Total cellular CXCR3 MFI levels were measured by flow cytometric analysis of fixed and permeabilized peripheral blood mononuclear cells (PBMCs) from MDD patients and matched controls ( n = 36). Stained PBMCs were gated on live CD3 + lymphocytes (T cells), CD4 + and CD8 + T cell subsets as well as CD3 − lymphocytes (non-T cells). Graphs depict medians with interquartile ranges. For all comparisons, the Wilcoxon signed-rank test was used. CXCR3: CXC-chemokine receptor type 3; CCR6: CC-chemokine receptor type 6; MFI: median fluorescence intensity.

    Article Snippet: Cell Purification For qRT-PCR and TCR sequencing analyses, CD4+ T cells were purified using magnetic beads (negative selection by BD IMag Human CD4 T Lymphocyte Enrichment Set, BD Biosciences) from PBMC aliquots thawed in cell separation buffer (1% human serum, 2 mM EDTA in PBS).

    Techniques: Staining, Expressing, Fluorescence, Concentration Assay, Flow Cytometry

    Regulatory T cells in major depressive disorder (MDD) patients and non-depressed controls. (A) Regulatory T cells (Tregs) were identified by flow cytometric analysis of peripheral blood mononuclear cells from MDD patients and matched non-depressed controls (CTR). Displayed values are frequencies of Tregs expressed as a percentage of live CD4 + T cells from a representative case–control pair. (B) Differences in Treg frequency are depicted for the entire cohort ( n = 40). (C) Negatively selected CD4 + T cells from a subsample of patients and matched controls ( n = 20) were analyzed for mRNA expression of the T helper-associated transcription factors Forkhead box P3 ( FOXP3 ), T-box 21 ( T-bet ), GATA binding protein 3 ( GATA3 ), and RAR related orphan receptor C ( RORC ), respectively. Expression was normalized to the geometric mean expression of three housekeeping genes ( IPO8, TBP, RPL13A ). (D) The correlation between the expression levels of the gene FOXP3 in purified CD4 + T cells and the frequency of Tregs expressed as a percentage of CD4 + T cells is plotted ( n = 20). Graphs depict medians with interquartile ranges. For all comparisons, the Wilcoxon signed-rank test was used.

    Journal: Frontiers in Immunology

    Article Title: T Cell Phenotype and T Cell Receptor Repertoire in Patients with Major Depressive Disorder

    doi: 10.3389/fimmu.2018.00291

    Figure Lengend Snippet: Regulatory T cells in major depressive disorder (MDD) patients and non-depressed controls. (A) Regulatory T cells (Tregs) were identified by flow cytometric analysis of peripheral blood mononuclear cells from MDD patients and matched non-depressed controls (CTR). Displayed values are frequencies of Tregs expressed as a percentage of live CD4 + T cells from a representative case–control pair. (B) Differences in Treg frequency are depicted for the entire cohort ( n = 40). (C) Negatively selected CD4 + T cells from a subsample of patients and matched controls ( n = 20) were analyzed for mRNA expression of the T helper-associated transcription factors Forkhead box P3 ( FOXP3 ), T-box 21 ( T-bet ), GATA binding protein 3 ( GATA3 ), and RAR related orphan receptor C ( RORC ), respectively. Expression was normalized to the geometric mean expression of three housekeeping genes ( IPO8, TBP, RPL13A ). (D) The correlation between the expression levels of the gene FOXP3 in purified CD4 + T cells and the frequency of Tregs expressed as a percentage of CD4 + T cells is plotted ( n = 20). Graphs depict medians with interquartile ranges. For all comparisons, the Wilcoxon signed-rank test was used.

    Article Snippet: Cell Purification For qRT-PCR and TCR sequencing analyses, CD4+ T cells were purified using magnetic beads (negative selection by BD IMag Human CD4 T Lymphocyte Enrichment Set, BD Biosciences) from PBMC aliquots thawed in cell separation buffer (1% human serum, 2 mM EDTA in PBS).

    Techniques: Flow Cytometry, Expressing, Binding Assay, Purification

    Peripheral blood counts and frequencies of major leukocyte subsets. (A) Absolute peripheral blood granulocyte, monocyte, and lymphocyte counts were obtained from major depressive disorder (MDD) patients and matched non-depressed controls (CTR) using a Coulter Ac·T Diff hematology analyzer ( n = 40). (B) Frequencies of total CD3 + lymphocytes (T cells), CD3 − CD56 − CD19 + B cells and CD3 − CD19 − CD20 − CD14 − CD56 + natural killer (NK) cells were obtained by flow cytometric analysis of thawed peripheral blood mononuclear cells. (C) T cells were further discriminated into CD4 + CD8 − and CD8 + CD4 − subsets. (D) Among NK cells, CD56 low CD16 + cytotoxic (NKc) cells and CD56 high CD16 − regulatory (NKreg) cells were also identified. Graphs depict medians with interquartile ranges. For all comparisons, the Wilcoxon signed-rank test was used.

    Journal: Frontiers in Immunology

    Article Title: T Cell Phenotype and T Cell Receptor Repertoire in Patients with Major Depressive Disorder

    doi: 10.3389/fimmu.2018.00291

    Figure Lengend Snippet: Peripheral blood counts and frequencies of major leukocyte subsets. (A) Absolute peripheral blood granulocyte, monocyte, and lymphocyte counts were obtained from major depressive disorder (MDD) patients and matched non-depressed controls (CTR) using a Coulter Ac·T Diff hematology analyzer ( n = 40). (B) Frequencies of total CD3 + lymphocytes (T cells), CD3 − CD56 − CD19 + B cells and CD3 − CD19 − CD20 − CD14 − CD56 + natural killer (NK) cells were obtained by flow cytometric analysis of thawed peripheral blood mononuclear cells. (C) T cells were further discriminated into CD4 + CD8 − and CD8 + CD4 − subsets. (D) Among NK cells, CD56 low CD16 + cytotoxic (NKc) cells and CD56 high CD16 − regulatory (NKreg) cells were also identified. Graphs depict medians with interquartile ranges. For all comparisons, the Wilcoxon signed-rank test was used.

    Article Snippet: Cell Purification For qRT-PCR and TCR sequencing analyses, CD4+ T cells were purified using magnetic beads (negative selection by BD IMag Human CD4 T Lymphocyte Enrichment Set, BD Biosciences) from PBMC aliquots thawed in cell separation buffer (1% human serum, 2 mM EDTA in PBS).

    Techniques: Flow Cytometry

    Signal scatter plots comparing the intensities of gene expressions on microarray chips within Ficoll and CPT-derived PBMC and their immune cell subsets for each healthy donor tested. The expression signals of Ficoll-derived cells were plotted against the expression signals of CPT-derived cells for each cell type and donor. A line of perfect correlation is indicated in all plots. Pearson’s correlation coefficients between each pair of samples are shown at the bottom right of each plot. The data showed that the pattern of gene expression was very similar between Ficoll and CPT-derived cell types, as indicated by the high correlation coefficients

    Journal: BMC Immunology

    Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

    doi: 10.1186/s12865-015-0113-0

    Figure Lengend Snippet: Signal scatter plots comparing the intensities of gene expressions on microarray chips within Ficoll and CPT-derived PBMC and their immune cell subsets for each healthy donor tested. The expression signals of Ficoll-derived cells were plotted against the expression signals of CPT-derived cells for each cell type and donor. A line of perfect correlation is indicated in all plots. Pearson’s correlation coefficients between each pair of samples are shown at the bottom right of each plot. The data showed that the pattern of gene expression was very similar between Ficoll and CPT-derived cell types, as indicated by the high correlation coefficients

    Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

    Techniques: Microarray, Cycling Probe Technology, Derivative Assay, Expressing

    Yield and viability of immune cell subsets prepared from PBMC isolated by either Ficoll or CPT protocol. a The mean number of CD19+, CD8+, CD14+, and CD4+ cells positively selected from Ficoll- and CPT-isolated PBMC collected from 6 healthy donors. The horizontal line denotes the means and each symbol represents an individual cell type isolated from Ficoll-PBMC (filled symbols) or CPT-PBMC (empty symbols) from each of 6 donors. b The mean viability of the same cell subsets. No significant differences ( P

    Journal: BMC Immunology

    Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

    doi: 10.1186/s12865-015-0113-0

    Figure Lengend Snippet: Yield and viability of immune cell subsets prepared from PBMC isolated by either Ficoll or CPT protocol. a The mean number of CD19+, CD8+, CD14+, and CD4+ cells positively selected from Ficoll- and CPT-isolated PBMC collected from 6 healthy donors. The horizontal line denotes the means and each symbol represents an individual cell type isolated from Ficoll-PBMC (filled symbols) or CPT-PBMC (empty symbols) from each of 6 donors. b The mean viability of the same cell subsets. No significant differences ( P

    Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

    Techniques: Isolation, Cycling Probe Technology

    Principle component analysis of gene expression showing distinct clusters between individual immune cell subsets but not between the same subsets derived from PBMC isolated by CPT or Ficoll protocols. Principle component analysis (PCA) was performed based on expression of all genes from the microarray. Individual immune cell types are represented by different colors: CD19+ B cells, blue; CD8+ T cells, green; CD14+ monocytes, purple; CD4+ T cells, orange; and total PBMC, red. The method used for PBMC isolation is represented by different symbols of differing size: CPT-based procedure by large circles and Ficoll-based procedure by small circles. PC#1, principle component #1; PC#2, principle component #2

    Journal: BMC Immunology

    Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

    doi: 10.1186/s12865-015-0113-0

    Figure Lengend Snippet: Principle component analysis of gene expression showing distinct clusters between individual immune cell subsets but not between the same subsets derived from PBMC isolated by CPT or Ficoll protocols. Principle component analysis (PCA) was performed based on expression of all genes from the microarray. Individual immune cell types are represented by different colors: CD19+ B cells, blue; CD8+ T cells, green; CD14+ monocytes, purple; CD4+ T cells, orange; and total PBMC, red. The method used for PBMC isolation is represented by different symbols of differing size: CPT-based procedure by large circles and Ficoll-based procedure by small circles. PC#1, principle component #1; PC#2, principle component #2

    Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

    Techniques: Expressing, Derivative Assay, Isolation, Cycling Probe Technology, Microarray

    Yield and quality of RNA recovered from total PBMC and their individual immune cell subsets did not significantly differ between PBMC isolated by Ficoll and CPT methods. a The mean amount of RNA per cell and ( b ) the mean RIN of RNA preparations extracted from total PBMC and their individual immune cell subsets derived from PBMC prepared by Ficoll and CPT methods from 6 health donors. There were no significant differences ( P

    Journal: BMC Immunology

    Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

    doi: 10.1186/s12865-015-0113-0

    Figure Lengend Snippet: Yield and quality of RNA recovered from total PBMC and their individual immune cell subsets did not significantly differ between PBMC isolated by Ficoll and CPT methods. a The mean amount of RNA per cell and ( b ) the mean RIN of RNA preparations extracted from total PBMC and their individual immune cell subsets derived from PBMC prepared by Ficoll and CPT methods from 6 health donors. There were no significant differences ( P

    Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

    Techniques: Isolation, Cycling Probe Technology, Derivative Assay

    Similar quantities of DNA with comparable quality were obtained from PBMC and immune cell subsets following either Ficoll or CPT protocol of PBMC isolation. a The mean amount of DNA extracted per cell and ( b ) the mean of the 260/280 ratios, as measure of DNA quality, were compared between DNA preparations obtained from total PBMC and immune cell subsets prepared from PBMC isolated by Ficoll and CPT methods from 6 healthy donors. There were no significant differences ( P

    Journal: BMC Immunology

    Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

    doi: 10.1186/s12865-015-0113-0

    Figure Lengend Snippet: Similar quantities of DNA with comparable quality were obtained from PBMC and immune cell subsets following either Ficoll or CPT protocol of PBMC isolation. a The mean amount of DNA extracted per cell and ( b ) the mean of the 260/280 ratios, as measure of DNA quality, were compared between DNA preparations obtained from total PBMC and immune cell subsets prepared from PBMC isolated by Ficoll and CPT methods from 6 healthy donors. There were no significant differences ( P

    Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

    Techniques: Cycling Probe Technology, Isolation

    Schematic outline of the study. PBMC from 6 healthy donors were isolated using Ficoll-Paque gradient fractionation or BD Vacutainer CPT protocol, and cell yield and purity were compared. Subsequently, immune cell subsets were separated by positive selection from PBMC obtained by both methods, and yields and viabilities of the subsets were determined and compared. RNA and DNA were extracted from total PBMC isolated by both protocols and from their subsets, and the nucleic acid yield and quality compared. Finally, gene expression analysis of individual immune cell subsets and total PBMC obtained by Ficoll and CPT isolation methods were performed and the results compared. LN 2 , liquid nitrogen

    Journal: BMC Immunology

    Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

    doi: 10.1186/s12865-015-0113-0

    Figure Lengend Snippet: Schematic outline of the study. PBMC from 6 healthy donors were isolated using Ficoll-Paque gradient fractionation or BD Vacutainer CPT protocol, and cell yield and purity were compared. Subsequently, immune cell subsets were separated by positive selection from PBMC obtained by both methods, and yields and viabilities of the subsets were determined and compared. RNA and DNA were extracted from total PBMC isolated by both protocols and from their subsets, and the nucleic acid yield and quality compared. Finally, gene expression analysis of individual immune cell subsets and total PBMC obtained by Ficoll and CPT isolation methods were performed and the results compared. LN 2 , liquid nitrogen

    Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

    Techniques: Isolation, Fractionation, Cycling Probe Technology, Selection, Expressing

    Yield and viability of PBMC isolated by the Ficoll and CPT protocols. a The mean numbers of PBMC per ml of blood obtained by Ficoll or CPT isolation procedure from 6 healthy donors. b The mean viability of PBMC freshly isolated from the same 6 healthy donors by either Ficoll gradient separation or CPT technique. No significant differences ( P

    Journal: BMC Immunology

    Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

    doi: 10.1186/s12865-015-0113-0

    Figure Lengend Snippet: Yield and viability of PBMC isolated by the Ficoll and CPT protocols. a The mean numbers of PBMC per ml of blood obtained by Ficoll or CPT isolation procedure from 6 healthy donors. b The mean viability of PBMC freshly isolated from the same 6 healthy donors by either Ficoll gradient separation or CPT technique. No significant differences ( P

    Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

    Techniques: Isolation, Cycling Probe Technology

    Purity of immune cell subsets obtained from PBMC isolated by either Ficoll or CPT procedures. CD19+, CD8+, CD14+, and CD4+ cell subsets were sequentially separated from PBMC isolated by Ficoll and CPT protocols from the same donor and their purity was determined by flow cytometry using antibodies against surface markers specific for individual immune cell types. Filled histograms were given by appropriate immunoglobulin isotype controls. Gates for determining positivity were established using isotype controls so that ~99.0 % of events were negative

    Journal: BMC Immunology

    Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

    doi: 10.1186/s12865-015-0113-0

    Figure Lengend Snippet: Purity of immune cell subsets obtained from PBMC isolated by either Ficoll or CPT procedures. CD19+, CD8+, CD14+, and CD4+ cell subsets were sequentially separated from PBMC isolated by Ficoll and CPT protocols from the same donor and their purity was determined by flow cytometry using antibodies against surface markers specific for individual immune cell types. Filled histograms were given by appropriate immunoglobulin isotype controls. Gates for determining positivity were established using isotype controls so that ~99.0 % of events were negative

    Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

    Techniques: Isolation, Cycling Probe Technology, Flow Cytometry, Cytometry

    Post-cryopreservation recovery and viability of PBMC isolated by the Ficoll and CPT methods. a The mean percent recovery after cryopreservation of PBMC collected using the Ficoll and CPT protocols. Percent recovery was calculated by dividing the number of PBMC recovered after thawing by the number of cells that were cryopreserved. b The mean viability of recovered PBMC isolated by the Ficoll and CPT technique. No significant differences ( P

    Journal: BMC Immunology

    Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

    doi: 10.1186/s12865-015-0113-0

    Figure Lengend Snippet: Post-cryopreservation recovery and viability of PBMC isolated by the Ficoll and CPT methods. a The mean percent recovery after cryopreservation of PBMC collected using the Ficoll and CPT protocols. Percent recovery was calculated by dividing the number of PBMC recovered after thawing by the number of cells that were cryopreserved. b The mean viability of recovered PBMC isolated by the Ficoll and CPT technique. No significant differences ( P

    Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

    Techniques: Isolation, Cycling Probe Technology