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Miltenyi Biotec peripheral blood mononuclear cells pbmcs
(a) Flow cytometry of Ficol separated human <t>PBMCs</t> labeled with <t>CD14</t> and CD3 demonstrated 17% of primary human PBMCs were CD14+ monocytes. (b) After isolation using magnetic bead separation (AutoMacs, Milteni) the purity of CD14+ monocytes was increased
Peripheral Blood Mononuclear Cells Pbmcs, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 93/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Implant debris particle size affects serum protein adsorption which may contribute to particle size-based bioreactivity differences"

Article Title: Implant debris particle size affects serum protein adsorption which may contribute to particle size-based bioreactivity differences

Journal: Journal of long-term effects of medical implants

doi:

(a) Flow cytometry of Ficol separated human PBMCs labeled with CD14 and CD3 demonstrated 17% of primary human PBMCs were CD14+ monocytes. (b) After isolation using magnetic bead separation (AutoMacs, Milteni) the purity of CD14+ monocytes was increased
Figure Legend Snippet: (a) Flow cytometry of Ficol separated human PBMCs labeled with CD14 and CD3 demonstrated 17% of primary human PBMCs were CD14+ monocytes. (b) After isolation using magnetic bead separation (AutoMacs, Milteni) the purity of CD14+ monocytes was increased

Techniques Used: Flow Cytometry, Cytometry, Labeling, Isolation

2) Product Images from "Preferential depletion of CD2low plasmacytoid dendritic cells in HIV-infected subjects"

Article Title: Preferential depletion of CD2low plasmacytoid dendritic cells in HIV-infected subjects

Journal: Cellular and Molecular Immunology

doi: 10.1038/cmi.2011.9

Characterization of CD2 low and CD2 high pDC subpopulations in HIV-infected subjects and healthy controls. ( a ) Total PBMCs were gated on forward and side scatter characteristics (left panel), and pDCs were identified as BDCA2 + 7-AAD − cells
Figure Legend Snippet: Characterization of CD2 low and CD2 high pDC subpopulations in HIV-infected subjects and healthy controls. ( a ) Total PBMCs were gated on forward and side scatter characteristics (left panel), and pDCs were identified as BDCA2 + 7-AAD − cells

Techniques Used: Infection

3) Product Images from "Estradiol and Progesterone Strongly Inhibit the Innate Immune Response of Mononuclear Cells in Newborns ▿"

Article Title: Estradiol and Progesterone Strongly Inhibit the Innate Immune Response of Mononuclear Cells in Newborns ▿

Journal: Infection and Immunity

doi: 10.1128/IAI.00076-11

The anti-inflammatory effect of estradiol and progesterone on CBMCs does not depend on modulation of TLR2, TLR4, and MYD88 expression. (A and B) Adult PBMCs (AD) and CBMCs (NB) ( n = 5 to 6) were incubated for 16 h with estradiol (E; 10 −7 M) or
Figure Legend Snippet: The anti-inflammatory effect of estradiol and progesterone on CBMCs does not depend on modulation of TLR2, TLR4, and MYD88 expression. (A and B) Adult PBMCs (AD) and CBMCs (NB) ( n = 5 to 6) were incubated for 16 h with estradiol (E; 10 −7 M) or

Techniques Used: Expressing, Incubation

CBMCs and newborn monocytes are more susceptible to the anti-inflammatory effect of estradiol and progesterone than adult PBMCs and monocytes. (A and B) PBMCs from adult males (black squares) and premenopausal females (black inverted triangles) and CBMCs
Figure Legend Snippet: CBMCs and newborn monocytes are more susceptible to the anti-inflammatory effect of estradiol and progesterone than adult PBMCs and monocytes. (A and B) PBMCs from adult males (black squares) and premenopausal females (black inverted triangles) and CBMCs

Techniques Used:

CBMCs express higher levels of estrogen and progesterone receptors than adult PBMCs and respond to specific receptor agonists. (A) Real-time PCR analysis of ER-α, ER-β, PR, PGRMC1, mPR-α, mPR-β, and mPR-γ mRNA levels
Figure Legend Snippet: CBMCs express higher levels of estrogen and progesterone receptors than adult PBMCs and respond to specific receptor agonists. (A) Real-time PCR analysis of ER-α, ER-β, PR, PGRMC1, mPR-α, mPR-β, and mPR-γ mRNA levels

Techniques Used: Real-time Polymerase Chain Reaction

4) Product Images from "High Specificity in Circulating Tumor Cell Identification Is Required for Accurate Evaluation of Programmed Death-Ligand 1"

Article Title: High Specificity in Circulating Tumor Cell Identification Is Required for Accurate Evaluation of Programmed Death-Ligand 1

Journal: PLoS ONE

doi: 10.1371/journal.pone.0159397

Flow cytometry reveals many CD11b + CD45 - cells are also pCK + . PBMCs isolated from patients with NSCLC and depleted of CD45 + cells were stained for flow cytometric evaluation. (A) Cells were gated first on live cell populations, then on cells which were either CD11b + CD45 lo or CD11b - CD45 - . (B) Subsequent plots displayed previously gated CD11b + CD45 lo cells (red) over all other live cells (black) on X and Y axes of FSC vs. SSC, (C) CD45 vs. pCK, or (E) CD45 vs. the isotype control for pCK, IgG1. (D) This staining and gating strategy was performed over three different patient samples to evaluate staining intensity, or mean fluorescence intensity (MFI), of pCK or (F) the isotype control. Significance was evaluated by one-tailed, paired T-tests with significance considered at p
Figure Legend Snippet: Flow cytometry reveals many CD11b + CD45 - cells are also pCK + . PBMCs isolated from patients with NSCLC and depleted of CD45 + cells were stained for flow cytometric evaluation. (A) Cells were gated first on live cell populations, then on cells which were either CD11b + CD45 lo or CD11b - CD45 - . (B) Subsequent plots displayed previously gated CD11b + CD45 lo cells (red) over all other live cells (black) on X and Y axes of FSC vs. SSC, (C) CD45 vs. pCK, or (E) CD45 vs. the isotype control for pCK, IgG1. (D) This staining and gating strategy was performed over three different patient samples to evaluate staining intensity, or mean fluorescence intensity (MFI), of pCK or (F) the isotype control. Significance was evaluated by one-tailed, paired T-tests with significance considered at p

Techniques Used: Flow Cytometry, Cytometry, Isolation, Staining, Fluorescence, One-tailed Test

5) Product Images from "A Selective Novel Peroxisome Proliferator–Activated Receptor (PPAR)-α Antagonist Induces Apoptosis and Inhibits Proliferation of CLL Cells In Vitro and In Vivo"

Article Title: A Selective Novel Peroxisome Proliferator–Activated Receptor (PPAR)-α Antagonist Induces Apoptosis and Inhibits Proliferation of CLL Cells In Vitro and In Vivo

Journal: Molecular Medicine

doi: 10.2119/molmed.2015.00139

CLL mouse model. (A) CLL PBMCs from two patients were labeled with CFSE and randomized among the groups. The 10 8 CFSE-labeled cells were injected intravenously into NSG mice (lacking T, B and NK cells). Groups of five mice received daily dosing of saline:
Figure Legend Snippet: CLL mouse model. (A) CLL PBMCs from two patients were labeled with CFSE and randomized among the groups. The 10 8 CFSE-labeled cells were injected intravenously into NSG mice (lacking T, B and NK cells). Groups of five mice received daily dosing of saline:

Techniques Used: Labeling, Injection, Mouse Assay

Related Articles

Cell Isolation:

Article Title: Expression of miR-34a in T-Cells Infected by Human T-Lymphotropic Virus 1
Article Snippet: .. CD4+ cells were isolated from normal peripheral blood mononuclear cells (PBMC) using the MACS CD4+ T cell Isolation Kit II or MACS CD4 Microbeads (Miltenyi Biotec). .. The resulting preparations contained ≥90% CD4+ cells measured by flow cytometry using a PE-labeled anti-CD4 antibody (Becton Dickinson).

Selection:

Article Title: Estradiol and Progesterone Strongly Inhibit the Innate Immune Response of Mononuclear Cells in Newborns ▿
Article Snippet: .. Monocytes were isolated from CBMCs and peripheral blood mononuclear cells (PBMCs) by positive selection using magnetic microbeads coupled to an anti-CD14 antibody (Miltenyi Biotec, Auburn, CA) with a purity of > 95%. ..

Article Title: Cholera Toxin Impairs the Differentiation of Monocytes into Dendritic Cells, Inducing Professional Antigen-Presenting Myeloid Cells ▿
Article Snippet: .. Human monocytes were purified from healthy donors' peripheral blood mononuclear cells (PBMC) by positive selection using anti-CD14-conjugated magnetic microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany). .. The recovered cells were 95% to 99% CD14+ as determined by flow cytometry using the FITC conjugate anti-human CD14 monoclonal antibody (MAb) (BD Biosciences, San Diego, CA).

Article Title: Implant debris particle size affects serum protein adsorption which may contribute to particle size-based bioreactivity differences
Article Snippet: .. Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll gradient and CD14+ monocytes were isolated from PBMCs by negative selection using magnetic bead antibody cocktail for CD3, CD7, CD16, CD19, CD56, CD123 and Glycophorin A (Miltenyi Biotec). .. Isolated human primary monocytes were assessed for > 90% purity using FACS.

Magnetic Beads:

Article Title: High Specificity in Circulating Tumor Cell Identification Is Required for Accurate Evaluation of Programmed Death-Ligand 1
Article Snippet: .. Peripheral blood mononuclear cells (PBMCs) were incubated with commercially available, pre-conjugated, anti-CD45 magnetic beads (Miltenyi) in a buffer containing 2 mM EDTA (Fisher Scientific) and 0.5% bovine serum albumin (BSA, Sigma-Aldrich) in phosphate buffered saline (PBS, Hyclone), after which CD45+ cells were removed using magnetic LS MACS columns (Miltenyi). .. Paramagnetic Particle (PMP) Preparation Streptavidin coupled PMPs (Dynabeads FlowComp Flexi, Invitrogen) at a concentration of 250 μg per reaction were used for all experiments.

Isolation:

Article Title: Estradiol and Progesterone Strongly Inhibit the Innate Immune Response of Mononuclear Cells in Newborns ▿
Article Snippet: .. Monocytes were isolated from CBMCs and peripheral blood mononuclear cells (PBMCs) by positive selection using magnetic microbeads coupled to an anti-CD14 antibody (Miltenyi Biotec, Auburn, CA) with a purity of > 95%. ..

Article Title: Cellular and humoral immune response to recombinant Escherichia coli OmpA in cows
Article Snippet: .. To ascertain that the production of IL-17A and IFN-γ was dependent on CD4+ T cells, peripheral blood mononuclear cells (PBMC) were isolated from two immunized cows and depleted of CD4+ cells by using MACS® beads (Miltenyi Biotech, Bergish Gladbach, Germany) as described [ ]. .. Percentages of CD4+ T cells were about 12% before and less than 1% after depletion, as assessed by flow cytometry as described [ ].

Article Title: Expression of miR-34a in T-Cells Infected by Human T-Lymphotropic Virus 1
Article Snippet: .. CD4+ cells were isolated from normal peripheral blood mononuclear cells (PBMC) using the MACS CD4+ T cell Isolation Kit II or MACS CD4 Microbeads (Miltenyi Biotec). .. The resulting preparations contained ≥90% CD4+ cells measured by flow cytometry using a PE-labeled anti-CD4 antibody (Becton Dickinson).

Article Title: An asthma-associated IL4R variant exacerbates airway inflammation by promoting conversion of regulatory T cells to TH17-like cells
Article Snippet: .. For iTreg cell differentiation, peripheral blood mononuclear cells (PBMC) were isolated and naïve CD4+ CDRA+ CDRO− T cells were derived by magnetic separation (Miltenyi biotech). .. The Isolated T cells were differentiated into iTreg differentiation as described for murine study and directly used for intracellular cytokine and FOXP3 expression analysis, using flow cytometry.

Article Title: Implant debris particle size affects serum protein adsorption which may contribute to particle size-based bioreactivity differences
Article Snippet: .. Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll gradient and CD14+ monocytes were isolated from PBMCs by negative selection using magnetic bead antibody cocktail for CD3, CD7, CD16, CD19, CD56, CD123 and Glycophorin A (Miltenyi Biotec). .. Isolated human primary monocytes were assessed for > 90% purity using FACS.

Purification:

Article Title: Cholera Toxin Impairs the Differentiation of Monocytes into Dendritic Cells, Inducing Professional Antigen-Presenting Myeloid Cells ▿
Article Snippet: .. Human monocytes were purified from healthy donors' peripheral blood mononuclear cells (PBMC) by positive selection using anti-CD14-conjugated magnetic microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany). .. The recovered cells were 95% to 99% CD14+ as determined by flow cytometry using the FITC conjugate anti-human CD14 monoclonal antibody (MAb) (BD Biosciences, San Diego, CA).

Incubation:

Article Title: High Specificity in Circulating Tumor Cell Identification Is Required for Accurate Evaluation of Programmed Death-Ligand 1
Article Snippet: .. Peripheral blood mononuclear cells (PBMCs) were incubated with commercially available, pre-conjugated, anti-CD45 magnetic beads (Miltenyi) in a buffer containing 2 mM EDTA (Fisher Scientific) and 0.5% bovine serum albumin (BSA, Sigma-Aldrich) in phosphate buffered saline (PBS, Hyclone), after which CD45+ cells were removed using magnetic LS MACS columns (Miltenyi). .. Paramagnetic Particle (PMP) Preparation Streptavidin coupled PMPs (Dynabeads FlowComp Flexi, Invitrogen) at a concentration of 250 μg per reaction were used for all experiments.

Magnetic Cell Separation:

Article Title: High Specificity in Circulating Tumor Cell Identification Is Required for Accurate Evaluation of Programmed Death-Ligand 1
Article Snippet: .. Peripheral blood mononuclear cells (PBMCs) were incubated with commercially available, pre-conjugated, anti-CD45 magnetic beads (Miltenyi) in a buffer containing 2 mM EDTA (Fisher Scientific) and 0.5% bovine serum albumin (BSA, Sigma-Aldrich) in phosphate buffered saline (PBS, Hyclone), after which CD45+ cells were removed using magnetic LS MACS columns (Miltenyi). .. Paramagnetic Particle (PMP) Preparation Streptavidin coupled PMPs (Dynabeads FlowComp Flexi, Invitrogen) at a concentration of 250 μg per reaction were used for all experiments.

Article Title: Cellular and humoral immune response to recombinant Escherichia coli OmpA in cows
Article Snippet: .. To ascertain that the production of IL-17A and IFN-γ was dependent on CD4+ T cells, peripheral blood mononuclear cells (PBMC) were isolated from two immunized cows and depleted of CD4+ cells by using MACS® beads (Miltenyi Biotech, Bergish Gladbach, Germany) as described [ ]. .. Percentages of CD4+ T cells were about 12% before and less than 1% after depletion, as assessed by flow cytometry as described [ ].

Article Title: Expression of miR-34a in T-Cells Infected by Human T-Lymphotropic Virus 1
Article Snippet: .. CD4+ cells were isolated from normal peripheral blood mononuclear cells (PBMC) using the MACS CD4+ T cell Isolation Kit II or MACS CD4 Microbeads (Miltenyi Biotec). .. The resulting preparations contained ≥90% CD4+ cells measured by flow cytometry using a PE-labeled anti-CD4 antibody (Becton Dickinson).

Staining:

Article Title: Preferential depletion of CD2low plasmacytoid dendritic cells in HIV-infected subjects
Article Snippet: .. Peripheral blood mononuclear cells (PBMCs) were stained with anti-BDCA2-FITC (Miltenyi, Bergisch Gladbach, Germany), anti-CD2-PE (Biolegend, San Diego, CA, USA) and 7-AAD. ..

Cell Differentiation:

Article Title: An asthma-associated IL4R variant exacerbates airway inflammation by promoting conversion of regulatory T cells to TH17-like cells
Article Snippet: .. For iTreg cell differentiation, peripheral blood mononuclear cells (PBMC) were isolated and naïve CD4+ CDRA+ CDRO− T cells were derived by magnetic separation (Miltenyi biotech). .. The Isolated T cells were differentiated into iTreg differentiation as described for murine study and directly used for intracellular cytokine and FOXP3 expression analysis, using flow cytometry.

Derivative Assay:

Article Title: An asthma-associated IL4R variant exacerbates airway inflammation by promoting conversion of regulatory T cells to TH17-like cells
Article Snippet: .. For iTreg cell differentiation, peripheral blood mononuclear cells (PBMC) were isolated and naïve CD4+ CDRA+ CDRO− T cells were derived by magnetic separation (Miltenyi biotech). .. The Isolated T cells were differentiated into iTreg differentiation as described for murine study and directly used for intracellular cytokine and FOXP3 expression analysis, using flow cytometry.

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    Miltenyi Biotec pbmcs
    SPM reversed the MGBG inhibitory effects on sOPN and CD16 expression in monocytes. <t>PBMCs</t> were cultured for 3 days with or without treatment. Cells were cultured with 10 μM MGBG and various concentrations of SPM. (A). MGBG significantly decreased sOPN; SPM reversed the MGBG inhibitory effects on sOPN. The average sOPN level of 3 day untreated and 10 μM MGBG treated cells was 30 and 2 ng/ml, respectively. n = 6, means and SEM. (B). MGBG inhibited monocyte CD16 expression; SPM reversed the MGBG inhibitory effects on CD16 expression. Cultured PBMCs were double stained with <t>CD14</t> and CD16 antibodies for flow cytometry analysis. The average CD16 geometric mean in untreated and MGBG treated CD14+ monocytes were measured at 510 and 249, respectively. n = 3, means and SEM. Data was presented as a percentage of inhibition of MGMG treatment only.
    Pbmcs, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 95/100, based on 3301 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmcs/product/Miltenyi Biotec
    Average 95 stars, based on 3301 article reviews
    Price from $9.99 to $1999.99
    pbmcs - by Bioz Stars, 2020-09
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    Miltenyi Biotec pha pbmcs
    AZT and ddI susceptibility of HIV-1 subtype E virus isolates from a family. (A) Replication kinetics. <t>PHA-PBMCs</t> were infected with virus stock (100 TCID 50 ) in the culture medium was monitored at the indicated time points. (B) Dose-response curve. Infected cells were cultured in the indicated concentrations of AZT (left panel) or ddI (right panel). RT activity at the peak of infection is expressed as a percentage of that of the untreated culture. Mean values with the standard errors of the three independent experiments for AZT and mean values of quadruplicates of an experiment for ddI are plotted.
    Pha Pbmcs, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pha pbmcs/product/Miltenyi Biotec
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    85
    Miltenyi Biotec lr eoc pbmcs
    Proliferation of Vγ9Vδ2 <t>PBMCs</t> from <t>EOC</t> patients and from healthy female donors following BrHPP or zoledronate stimulation. Vδ2 + CD3 + cell frequencies among expanded cells ( A ) and Vδ2 + CD3 + cell numbers generated from 1×10 6 PBMCs ( B ) were measured at 14 days after BrHPP ( ▴ ) or zoledronate (Zol) ( ▪ ) stimulation. The results from n = 60 patient PBMCs and n = 13 donor PBMCs are shown.
    Lr Eoc Pbmcs, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lr eoc pbmcs/product/Miltenyi Biotec
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    SPM reversed the MGBG inhibitory effects on sOPN and CD16 expression in monocytes. PBMCs were cultured for 3 days with or without treatment. Cells were cultured with 10 μM MGBG and various concentrations of SPM. (A). MGBG significantly decreased sOPN; SPM reversed the MGBG inhibitory effects on sOPN. The average sOPN level of 3 day untreated and 10 μM MGBG treated cells was 30 and 2 ng/ml, respectively. n = 6, means and SEM. (B). MGBG inhibited monocyte CD16 expression; SPM reversed the MGBG inhibitory effects on CD16 expression. Cultured PBMCs were double stained with CD14 and CD16 antibodies for flow cytometry analysis. The average CD16 geometric mean in untreated and MGBG treated CD14+ monocytes were measured at 510 and 249, respectively. n = 3, means and SEM. Data was presented as a percentage of inhibition of MGMG treatment only.

    Journal: PLoS ONE

    Article Title: Methylglyoxal-bis-guanylhydrazone inhibits osteopontin expression and differentiation in cultured human monocytes

    doi: 10.1371/journal.pone.0192680

    Figure Lengend Snippet: SPM reversed the MGBG inhibitory effects on sOPN and CD16 expression in monocytes. PBMCs were cultured for 3 days with or without treatment. Cells were cultured with 10 μM MGBG and various concentrations of SPM. (A). MGBG significantly decreased sOPN; SPM reversed the MGBG inhibitory effects on sOPN. The average sOPN level of 3 day untreated and 10 μM MGBG treated cells was 30 and 2 ng/ml, respectively. n = 6, means and SEM. (B). MGBG inhibited monocyte CD16 expression; SPM reversed the MGBG inhibitory effects on CD16 expression. Cultured PBMCs were double stained with CD14 and CD16 antibodies for flow cytometry analysis. The average CD16 geometric mean in untreated and MGBG treated CD14+ monocytes were measured at 510 and 249, respectively. n = 3, means and SEM. Data was presented as a percentage of inhibition of MGMG treatment only.

    Article Snippet: Monocytes were isolated from PBMCs using anti-CD14 (Miltenyi Biotech, Auburn, CA) MACS magnetic separation system.

    Techniques: Expressing, Cell Culture, Staining, Flow Cytometry, Cytometry, Inhibition

    MGBG showed greater inhibition on CD16 expression in monocytes than that in differentiated macrophages. Monocytes were isolated from PBMCs using CD14 microbeads. Cells were cultured for 1, 3, and 6 days with various concentrations of MGBG, and collected for CD16 expression measurement using flow cytometry. MGBG showed greater inhibitory effect on CD 16 expression in 1 and 3 day monocytes than that in differentiated macrophages. The average CD16 geometric mean fluorescence was measured at 108, 448, and 249 for 1, 3, and 6 day untreated cells, respectively. n = 4, means and SEM.

    Journal: PLoS ONE

    Article Title: Methylglyoxal-bis-guanylhydrazone inhibits osteopontin expression and differentiation in cultured human monocytes

    doi: 10.1371/journal.pone.0192680

    Figure Lengend Snippet: MGBG showed greater inhibition on CD16 expression in monocytes than that in differentiated macrophages. Monocytes were isolated from PBMCs using CD14 microbeads. Cells were cultured for 1, 3, and 6 days with various concentrations of MGBG, and collected for CD16 expression measurement using flow cytometry. MGBG showed greater inhibitory effect on CD 16 expression in 1 and 3 day monocytes than that in differentiated macrophages. The average CD16 geometric mean fluorescence was measured at 108, 448, and 249 for 1, 3, and 6 day untreated cells, respectively. n = 4, means and SEM.

    Article Snippet: Monocytes were isolated from PBMCs using anti-CD14 (Miltenyi Biotech, Auburn, CA) MACS magnetic separation system.

    Techniques: Inhibition, Expressing, Isolation, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

    HER-2/neu p373–382-specific CD8 + T cells have increased recognition of human breast cancer cell lines (A–B) Shown are the mean (±SEM, n=3) numbers of ELISpots obtained in response of pFLU, p369–377, or p373–382 peptide-specific CD8 + T cells derived from short-term culture to (A) autologous PBMC pulsed with pFLU, p369–377, or p373–382 or (B) irradiated tumor cells. Results are from a single donor and are representative of similar results obtained with 3 other donors. (C) Shown are the mean (±SEM, n=4) % lysis values calculated from the impedance-based lysis assay at 10 hours following the addition of T cells. Also shown are the expression levels of HER-2/neu and HLA-A2 genotyping results for each of the cell lines. Results are pertinent to panels B and C. HER-2/neu overexpression levels in each cell line and HLA-A2 expression are indicated below the x-axis. Expression levels were determined by flow cytometry and RT-PCR. p-values in panels B and C were calculated using an unpaired Student's t-test. (D) Shown are the mean (±SEM, n=3) numbers of IFN-γ ELISpots obtained in response to pFLU or p373–382 peptide-specific CD8 + T cells derived from short-term culture to autologous PBMC pulsed with pFLU, p373–382, HER-2/neu ECD or ovalbumin protein. Experiment was repeated twice in triplicate with both experiments yielding similar results.

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

    Article Title: Enzymatic discovery of a HER-2/neu epitope that generates cross-reactive T cells 1

    doi: 10.4049/jimmunol.1201264

    Figure Lengend Snippet: HER-2/neu p373–382-specific CD8 + T cells have increased recognition of human breast cancer cell lines (A–B) Shown are the mean (±SEM, n=3) numbers of ELISpots obtained in response of pFLU, p369–377, or p373–382 peptide-specific CD8 + T cells derived from short-term culture to (A) autologous PBMC pulsed with pFLU, p369–377, or p373–382 or (B) irradiated tumor cells. Results are from a single donor and are representative of similar results obtained with 3 other donors. (C) Shown are the mean (±SEM, n=4) % lysis values calculated from the impedance-based lysis assay at 10 hours following the addition of T cells. Also shown are the expression levels of HER-2/neu and HLA-A2 genotyping results for each of the cell lines. Results are pertinent to panels B and C. HER-2/neu overexpression levels in each cell line and HLA-A2 expression are indicated below the x-axis. Expression levels were determined by flow cytometry and RT-PCR. p-values in panels B and C were calculated using an unpaired Student's t-test. (D) Shown are the mean (±SEM, n=3) numbers of IFN-γ ELISpots obtained in response to pFLU or p373–382 peptide-specific CD8 + T cells derived from short-term culture to autologous PBMC pulsed with pFLU, p373–382, HER-2/neu ECD or ovalbumin protein. Experiment was repeated twice in triplicate with both experiments yielding similar results.

    Article Snippet: PBMC samples which stained positive for HLA-A2 were further enriched with the CD8+ T Cell Isolation Kit II (Miltenyi Biotec) and separated by AutoMACS (Miltenyi Biotec) per the manufacturer's protocol.

    Techniques: Derivative Assay, Irradiation, Lysis, Expressing, Over Expression, Flow Cytometry, Cytometry, Reverse Transcription Polymerase Chain Reaction

    IFN-γ and lytic responses of p373–382-generated T cells are HLA class I restricted (A–B) Shown are the mean (±SEM, n=3) numbers of p373–382-specific ELISpots obtained in response to p369–377 or p373–382 peptide-specific CD8 + T cells derived from short-term culture to autologous PBMC pulsed with p369–377 or p373–382 in the presence or absence of (A) HLA-ABC blocking antibody or (B) HLA-A2 blocking antibody. One representative experiment of three is shown. (C) Shown are the mean (±SEM, n=4) % lysis values calculated from the impedance-based lysis assay at 10 hours following the addition of T cells. The target cells were pretreated with or without 10 μg/ml HLA-ABC blocking antibody or HLA-A2 blocking antibody and appropriate isotype control antibodies 1 hour prior to T cell addition. Similar results were obtained with T cells generated from 3 other donors. p-values were calculated using a non-parametric Mann-Whitney Test.

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

    Article Title: Enzymatic discovery of a HER-2/neu epitope that generates cross-reactive T cells 1

    doi: 10.4049/jimmunol.1201264

    Figure Lengend Snippet: IFN-γ and lytic responses of p373–382-generated T cells are HLA class I restricted (A–B) Shown are the mean (±SEM, n=3) numbers of p373–382-specific ELISpots obtained in response to p369–377 or p373–382 peptide-specific CD8 + T cells derived from short-term culture to autologous PBMC pulsed with p369–377 or p373–382 in the presence or absence of (A) HLA-ABC blocking antibody or (B) HLA-A2 blocking antibody. One representative experiment of three is shown. (C) Shown are the mean (±SEM, n=4) % lysis values calculated from the impedance-based lysis assay at 10 hours following the addition of T cells. The target cells were pretreated with or without 10 μg/ml HLA-ABC blocking antibody or HLA-A2 blocking antibody and appropriate isotype control antibodies 1 hour prior to T cell addition. Similar results were obtained with T cells generated from 3 other donors. p-values were calculated using a non-parametric Mann-Whitney Test.

    Article Snippet: PBMC samples which stained positive for HLA-A2 were further enriched with the CD8+ T Cell Isolation Kit II (Miltenyi Biotec) and separated by AutoMACS (Miltenyi Biotec) per the manufacturer's protocol.

    Techniques: Generated, Derivative Assay, Blocking Assay, Lysis, MANN-WHITNEY

    AZT and ddI susceptibility of HIV-1 subtype E virus isolates from a family. (A) Replication kinetics. PHA-PBMCs were infected with virus stock (100 TCID 50 ) in the culture medium was monitored at the indicated time points. (B) Dose-response curve. Infected cells were cultured in the indicated concentrations of AZT (left panel) or ddI (right panel). RT activity at the peak of infection is expressed as a percentage of that of the untreated culture. Mean values with the standard errors of the three independent experiments for AZT and mean values of quadruplicates of an experiment for ddI are plotted.

    Journal: Journal of Virology

    Article Title: Convergent Evolution of Reverse Transcriptase (RT) Genes of Human Immunodeficiency Virus Type 1 Subtypes E and B following Nucleoside Analogue RT Inhibitor Therapies

    doi:

    Figure Lengend Snippet: AZT and ddI susceptibility of HIV-1 subtype E virus isolates from a family. (A) Replication kinetics. PHA-PBMCs were infected with virus stock (100 TCID 50 ) in the culture medium was monitored at the indicated time points. (B) Dose-response curve. Infected cells were cultured in the indicated concentrations of AZT (left panel) or ddI (right panel). RT activity at the peak of infection is expressed as a percentage of that of the untreated culture. Mean values with the standard errors of the three independent experiments for AZT and mean values of quadruplicates of an experiment for ddI are plotted.

    Article Snippet: Because HIV-1NH2 and HIV-1NH3 replicated poorly in the PHA-PBMCs, PBMCs were depleted of CD8+ cells with anti-CD8 antibody and a magnetic cell sorter (Miltenyi Biotec, Bergisch-Gladbach, Germany), and the CD8-positive-cell-depleted PHA-PBMCs were used for the present assay.

    Techniques: Infection, Cell Culture, Activity Assay

    Proliferation of Vγ9Vδ2 PBMCs from EOC patients and from healthy female donors following BrHPP or zoledronate stimulation. Vδ2 + CD3 + cell frequencies among expanded cells ( A ) and Vδ2 + CD3 + cell numbers generated from 1×10 6 PBMCs ( B ) were measured at 14 days after BrHPP ( ▴ ) or zoledronate (Zol) ( ▪ ) stimulation. The results from n = 60 patient PBMCs and n = 13 donor PBMCs are shown.

    Journal: PLoS ONE

    Article Title: A Quantitative Deficiency in Peripheral Blood V?9V?2 Cells Is a Negative Prognostic Biomarker in Ovarian Cancer Patients

    doi: 10.1371/journal.pone.0063322

    Figure Lengend Snippet: Proliferation of Vγ9Vδ2 PBMCs from EOC patients and from healthy female donors following BrHPP or zoledronate stimulation. Vδ2 + CD3 + cell frequencies among expanded cells ( A ) and Vδ2 + CD3 + cell numbers generated from 1×10 6 PBMCs ( B ) were measured at 14 days after BrHPP ( ▴ ) or zoledronate (Zol) ( ▪ ) stimulation. The results from n = 60 patient PBMCs and n = 13 donor PBMCs are shown.

    Article Snippet: In additional assays, we tested an oblique method of cell depletion to remove Tregs from LR EOC PBMCs while preserving CD25+ activated cells with the use of the CD4+ CD25+ CD127dim/− Regulatory T Cell Isolation Kit II from Miltenyi Biotec (data not shown).

    Techniques: Generated

    Disease-free survival of chemotherapy-treated EOC patients according to Vγ9Vδ2 cell frequencies among ex vivo PBMCs. Disease-free survival Kaplan-Meier curves of patients with Vγ9Vδ2 cell frequencies in PBMCs of 0.35% or less (≤0.35%) (n = 9) or greater than 0.35% ( > 0.35%) (n = 43) at the time of blood collection. p value

    Journal: PLoS ONE

    Article Title: A Quantitative Deficiency in Peripheral Blood V?9V?2 Cells Is a Negative Prognostic Biomarker in Ovarian Cancer Patients

    doi: 10.1371/journal.pone.0063322

    Figure Lengend Snippet: Disease-free survival of chemotherapy-treated EOC patients according to Vγ9Vδ2 cell frequencies among ex vivo PBMCs. Disease-free survival Kaplan-Meier curves of patients with Vγ9Vδ2 cell frequencies in PBMCs of 0.35% or less (≤0.35%) (n = 9) or greater than 0.35% ( > 0.35%) (n = 43) at the time of blood collection. p value

    Article Snippet: In additional assays, we tested an oblique method of cell depletion to remove Tregs from LR EOC PBMCs while preserving CD25+ activated cells with the use of the CD4+ CD25+ CD127dim/− Regulatory T Cell Isolation Kit II from Miltenyi Biotec (data not shown).

    Techniques: Ex Vivo

    A reduced naive Vγ9Vδ2 subset in ex vivo LR EOC PBMCs. Percentages of naive (CD27 + CD45RA + ), central memory (CM) (CD27 + CD45RA − ), effector memory (EM) (CD27 − CD45RA − ) and terminally differentiated effector memory (TEMRA) (CD27 − CD45RA + ) cells among the Vδ2 + CD3 + cells in ex vivo LR (n = 13) and R (n = 11) EOC PBMCs.

    Journal: PLoS ONE

    Article Title: A Quantitative Deficiency in Peripheral Blood V?9V?2 Cells Is a Negative Prognostic Biomarker in Ovarian Cancer Patients

    doi: 10.1371/journal.pone.0063322

    Figure Lengend Snippet: A reduced naive Vγ9Vδ2 subset in ex vivo LR EOC PBMCs. Percentages of naive (CD27 + CD45RA + ), central memory (CM) (CD27 + CD45RA − ), effector memory (EM) (CD27 − CD45RA − ) and terminally differentiated effector memory (TEMRA) (CD27 − CD45RA + ) cells among the Vδ2 + CD3 + cells in ex vivo LR (n = 13) and R (n = 11) EOC PBMCs.

    Article Snippet: In additional assays, we tested an oblique method of cell depletion to remove Tregs from LR EOC PBMCs while preserving CD25+ activated cells with the use of the CD4+ CD25+ CD127dim/− Regulatory T Cell Isolation Kit II from Miltenyi Biotec (data not shown).

    Techniques: Ex Vivo

    Reduced functionality of Vγ9Vδ2 cells in LR EOC PBMCs without defects in expression level of CD3ζ and CRTAM molecules. A, B ) Vδ2 + CD3 + cell fold increases in LR (n = 16) and R (n = 32) EOC PBMCs at 14 days after BrHPP ( A ) or Zol ( B ) stimulation. C ) Percentages of IFN-γ + cells in the Vδ2 + CD3 + cells measured at 5 h after the stimulation of ex vivo LR (n = 10) and R (n = 10) PBMCs with BrHPP (3 µM) or PMA/ionomycin (PMA/iono). D ) Percentages of IFN-γ + cells ( left panel ) and TNF-α + cells ( right panel ) among the Vδ2 + CD3 + cells measured at 5 h after stimulations of ex vivo LR (n = 4) and R (n = 4) PBMCs with increasing doses of BrHPP (0.1 to 6 µM). E ) Expression of the CD3ζ chain measured on Vγ9Vδ2 PBMCs from the LR and R groups. Percentage ( upper panel ) and MFI ( lower panel ) of CD3ζ staining in the Vδ2 + CD3 + cells from LR EOC PBMCs (n = 5), R EOC PBMCs (n = 3) and R donor PBMCs (n = 3). F ) CRTAM expression on Vγ9Vδ2 cells measured at 20 h after stimulation of LR (n = 8) and R (n = 8) EOC PBMCs with BrHPP. No stim: no stimulation.

    Journal: PLoS ONE

    Article Title: A Quantitative Deficiency in Peripheral Blood V?9V?2 Cells Is a Negative Prognostic Biomarker in Ovarian Cancer Patients

    doi: 10.1371/journal.pone.0063322

    Figure Lengend Snippet: Reduced functionality of Vγ9Vδ2 cells in LR EOC PBMCs without defects in expression level of CD3ζ and CRTAM molecules. A, B ) Vδ2 + CD3 + cell fold increases in LR (n = 16) and R (n = 32) EOC PBMCs at 14 days after BrHPP ( A ) or Zol ( B ) stimulation. C ) Percentages of IFN-γ + cells in the Vδ2 + CD3 + cells measured at 5 h after the stimulation of ex vivo LR (n = 10) and R (n = 10) PBMCs with BrHPP (3 µM) or PMA/ionomycin (PMA/iono). D ) Percentages of IFN-γ + cells ( left panel ) and TNF-α + cells ( right panel ) among the Vδ2 + CD3 + cells measured at 5 h after stimulations of ex vivo LR (n = 4) and R (n = 4) PBMCs with increasing doses of BrHPP (0.1 to 6 µM). E ) Expression of the CD3ζ chain measured on Vγ9Vδ2 PBMCs from the LR and R groups. Percentage ( upper panel ) and MFI ( lower panel ) of CD3ζ staining in the Vδ2 + CD3 + cells from LR EOC PBMCs (n = 5), R EOC PBMCs (n = 3) and R donor PBMCs (n = 3). F ) CRTAM expression on Vγ9Vδ2 cells measured at 20 h after stimulation of LR (n = 8) and R (n = 8) EOC PBMCs with BrHPP. No stim: no stimulation.

    Article Snippet: In additional assays, we tested an oblique method of cell depletion to remove Tregs from LR EOC PBMCs while preserving CD25+ activated cells with the use of the CD4+ CD25+ CD127dim/− Regulatory T Cell Isolation Kit II from Miltenyi Biotec (data not shown).

    Techniques: Expressing, Ex Vivo, Staining

    An imbalanced ratio of Tregs and Vγ9Vδ2 cells in ex vivo LR EOC PBMCs. A ) CD4 + CD25 high FoxP3 high cell (Treg) frequencies and corresponding Vδ2 + CD3 + cell frequencies in ex vivo LR (n = 13) and R (n = 11) PBMCs. B ) The ratios of Tregs (%) to Vδ2 + CD3 + cells (%) (Treg:Vδ2 + ratio) among PBMCs in the LR and R groups. C ) LR PBMCs from EOC patients (n = 6) were stimulated with BrHPP (▴) or Zol (▪) and IL-2 before and after the depletion of CD25 + cells; the proliferation of Vδ2 + CD3 + cells was analyzed on day 7. Vδ2 + CD3 + cell frequencies among expanded cells ( left panel ) and Vδ2 + CD3 + cell fold increases ( right panel ) are shown.

    Journal: PLoS ONE

    Article Title: A Quantitative Deficiency in Peripheral Blood V?9V?2 Cells Is a Negative Prognostic Biomarker in Ovarian Cancer Patients

    doi: 10.1371/journal.pone.0063322

    Figure Lengend Snippet: An imbalanced ratio of Tregs and Vγ9Vδ2 cells in ex vivo LR EOC PBMCs. A ) CD4 + CD25 high FoxP3 high cell (Treg) frequencies and corresponding Vδ2 + CD3 + cell frequencies in ex vivo LR (n = 13) and R (n = 11) PBMCs. B ) The ratios of Tregs (%) to Vδ2 + CD3 + cells (%) (Treg:Vδ2 + ratio) among PBMCs in the LR and R groups. C ) LR PBMCs from EOC patients (n = 6) were stimulated with BrHPP (▴) or Zol (▪) and IL-2 before and after the depletion of CD25 + cells; the proliferation of Vδ2 + CD3 + cells was analyzed on day 7. Vδ2 + CD3 + cell frequencies among expanded cells ( left panel ) and Vδ2 + CD3 + cell fold increases ( right panel ) are shown.

    Article Snippet: In additional assays, we tested an oblique method of cell depletion to remove Tregs from LR EOC PBMCs while preserving CD25+ activated cells with the use of the CD4+ CD25+ CD127dim/− Regulatory T Cell Isolation Kit II from Miltenyi Biotec (data not shown).

    Techniques: Ex Vivo

    Specific quantitative deficiencies of Vγ9Vδ2 cells in ex vivo LR EOC PBMCs. Frequencies of Vδ2 + CD3 + cells ( A ), Vδ2 − γδ + CD3 + cells ( B ) and CD3 + cells ( C ) among LR (n = 16), Br-LR (n = 8), Zol-LR (n = 4) and R (n = 32) EOC PBMCs are shown. Differences between LR group and each of other groups are indicated. A ) A receiver-operator characteristic (ROC) analysis was performed in which LR PBMC samples were compared to the other PBMC samples. Dashed lines indicate cut-offs at 0.35% and 0.8%. Sp: Specificity. Se: sensibility.

    Journal: PLoS ONE

    Article Title: A Quantitative Deficiency in Peripheral Blood V?9V?2 Cells Is a Negative Prognostic Biomarker in Ovarian Cancer Patients

    doi: 10.1371/journal.pone.0063322

    Figure Lengend Snippet: Specific quantitative deficiencies of Vγ9Vδ2 cells in ex vivo LR EOC PBMCs. Frequencies of Vδ2 + CD3 + cells ( A ), Vδ2 − γδ + CD3 + cells ( B ) and CD3 + cells ( C ) among LR (n = 16), Br-LR (n = 8), Zol-LR (n = 4) and R (n = 32) EOC PBMCs are shown. Differences between LR group and each of other groups are indicated. A ) A receiver-operator characteristic (ROC) analysis was performed in which LR PBMC samples were compared to the other PBMC samples. Dashed lines indicate cut-offs at 0.35% and 0.8%. Sp: Specificity. Se: sensibility.

    Article Snippet: In additional assays, we tested an oblique method of cell depletion to remove Tregs from LR EOC PBMCs while preserving CD25+ activated cells with the use of the CD4+ CD25+ CD127dim/− Regulatory T Cell Isolation Kit II from Miltenyi Biotec (data not shown).

    Techniques: Ex Vivo