primary peripheral blood mononuclear cells pbmc normal human  (ATCC)


Bioz Verified Symbol ATCC is a verified supplier
Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    Name:
    Primary Peripheral Blood Mononuclear Cells PBMC Normal Human
    Description:
    Applications Applications for use include the study of immunology infection cancer hematology and t cell suppression assay Cell Type Mononuclear Host Homo sapiens human
    Catalog Number:
    pcs-800-011
    Price:
    None
    Applications:
    Applications for use include the study of immunology, infection, cancer, hematology, and t-cell suppression assay.
    Cell Type:
    Mononuclear
    Host:
    Homo sapiens, human
    Buy from Supplier


    Structured Review

    ATCC primary peripheral blood mononuclear cells pbmc normal human
    Measurements of the biological activity of desArg 9 BK released from AP41 bacteria and cell surface expression of B1R. A HEK293 cells stably transfected with B1R were treated with (i) buffer, (ii) the B1R antagonist DLKD, (iii) desArg 9 BK, (iv) a mixture of desArg 9 BK and DLKD, (v) BK cleavage products released from AP41 bacteria, and (vi) a mixture of BK cleavage products released from AP41 bacteria and DLKD. Inositol phosphates were detected as described in Materials and Methods. The result is representative of three experiments with each point performed in duplicates. B IMR-90 cells were incubated for 6 h with (i) medium, (ii) <t>PBMC</t> exudates (monocytic cells that had been stimulated for 24 h with 1% S. pyogenes overnight culture supernatants), or (iii) PBMC exudates in the presence of 10 µM desArg 9 BK. All treatments were performed in MEM medium and in the absence of serum and antibiotics. After intensive washing, cells were assayed for specific [ 3 H]des-Arg 10 kallidin (B1R ligand) binding. Binding of [ 3 H]des-Arg 10 kallidin to non-stimulated cells (control) was normalized to 100% within each experiment. Results represent the mean ± standard deviation of 3 independent experiments performed in triplicates. * p
    Applications Applications for use include the study of immunology infection cancer hematology and t cell suppression assay Cell Type Mononuclear Host Homo sapiens human
    https://www.bioz.com/result/primary peripheral blood mononuclear cells pbmc normal human/product/ATCC
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    primary peripheral blood mononuclear cells pbmc normal human - by Bioz Stars, 2021-03
    95/100 stars

    Images

    1) Product Images from "Activation of TAFI on the Surface of Streptococcus pyogenes Evokes Inflammatory Reactions by Modulating the Kallikrein/Kinin System"

    Article Title: Activation of TAFI on the Surface of Streptococcus pyogenes Evokes Inflammatory Reactions by Modulating the Kallikrein/Kinin System

    Journal: Journal of Innate Immunity

    doi: 10.1159/000145543

    Measurements of the biological activity of desArg 9 BK released from AP41 bacteria and cell surface expression of B1R. A HEK293 cells stably transfected with B1R were treated with (i) buffer, (ii) the B1R antagonist DLKD, (iii) desArg 9 BK, (iv) a mixture of desArg 9 BK and DLKD, (v) BK cleavage products released from AP41 bacteria, and (vi) a mixture of BK cleavage products released from AP41 bacteria and DLKD. Inositol phosphates were detected as described in Materials and Methods. The result is representative of three experiments with each point performed in duplicates. B IMR-90 cells were incubated for 6 h with (i) medium, (ii) PBMC exudates (monocytic cells that had been stimulated for 24 h with 1% S. pyogenes overnight culture supernatants), or (iii) PBMC exudates in the presence of 10 µM desArg 9 BK. All treatments were performed in MEM medium and in the absence of serum and antibiotics. After intensive washing, cells were assayed for specific [ 3 H]des-Arg 10 kallidin (B1R ligand) binding. Binding of [ 3 H]des-Arg 10 kallidin to non-stimulated cells (control) was normalized to 100% within each experiment. Results represent the mean ± standard deviation of 3 independent experiments performed in triplicates. * p
    Figure Legend Snippet: Measurements of the biological activity of desArg 9 BK released from AP41 bacteria and cell surface expression of B1R. A HEK293 cells stably transfected with B1R were treated with (i) buffer, (ii) the B1R antagonist DLKD, (iii) desArg 9 BK, (iv) a mixture of desArg 9 BK and DLKD, (v) BK cleavage products released from AP41 bacteria, and (vi) a mixture of BK cleavage products released from AP41 bacteria and DLKD. Inositol phosphates were detected as described in Materials and Methods. The result is representative of three experiments with each point performed in duplicates. B IMR-90 cells were incubated for 6 h with (i) medium, (ii) PBMC exudates (monocytic cells that had been stimulated for 24 h with 1% S. pyogenes overnight culture supernatants), or (iii) PBMC exudates in the presence of 10 µM desArg 9 BK. All treatments were performed in MEM medium and in the absence of serum and antibiotics. After intensive washing, cells were assayed for specific [ 3 H]des-Arg 10 kallidin (B1R ligand) binding. Binding of [ 3 H]des-Arg 10 kallidin to non-stimulated cells (control) was normalized to 100% within each experiment. Results represent the mean ± standard deviation of 3 independent experiments performed in triplicates. * p

    Techniques Used: Activity Assay, Expressing, Stable Transfection, Transfection, Incubation, Ligand Binding Assay, Binding Assay, Standard Deviation

    2) Product Images from "Modeling Cell-Specific Dynamics and Regulation of the Common Gamma Chain Cytokines"

    Article Title: Modeling Cell-Specific Dynamics and Regulation of the Common Gamma Chain Cytokines

    Journal: bioRxiv

    doi: 10.1101/778894

    Tensor factorization to map model-predicted cytokine responses. A) Measured receptor abundance for ten PBMC-derived subpopulations. Points and error bars show geometric mean and standard deviation respectively (N = 4). Error bars for some points are too small to display. B-C) PCA scores (B) and loadings (C) of receptor abundance. Axis label percentages indicate percent variance explained. D) Schematic representation of CP decomposition. Model predictions are arranged in a cube depending upon the time, ligand treatment, and cell type being modeled. CP decomposition then helps to visualize this space. E) Percent variance reconstructed (R2X) versus the number of components used in non-negative CP decomposition. F-I) Component values versus time (F), cell type (G-H), or ligand stimulation (I). The variation explained by each component is the product of the component’s time, ligand, and cell type factorization. Ligand components with only negligible values (
    Figure Legend Snippet: Tensor factorization to map model-predicted cytokine responses. A) Measured receptor abundance for ten PBMC-derived subpopulations. Points and error bars show geometric mean and standard deviation respectively (N = 4). Error bars for some points are too small to display. B-C) PCA scores (B) and loadings (C) of receptor abundance. Axis label percentages indicate percent variance explained. D) Schematic representation of CP decomposition. Model predictions are arranged in a cube depending upon the time, ligand treatment, and cell type being modeled. CP decomposition then helps to visualize this space. E) Percent variance reconstructed (R2X) versus the number of components used in non-negative CP decomposition. F-I) Component values versus time (F), cell type (G-H), or ligand stimulation (I). The variation explained by each component is the product of the component’s time, ligand, and cell type factorization. Ligand components with only negligible values (

    Techniques Used: Derivative Assay, Standard Deviation

    Receptor quantification and gating of PBMC-derived immune cell types. A) Preliminary gating for single lyphocytes. B) Example staining for CD122 (red), the corresponding isotype control (blue), and unstained cells (black). C) Gating for live T helper and T regulatory cells during receptor quantification. D) Live cell NK cell gating. E) Live cell CD8+ T cell gating. F) Gating for fixed T helper and T regulatory cells during pSTAT5 quantification. G) Fixed CD8+ T cell and NK cell gating.
    Figure Legend Snippet: Receptor quantification and gating of PBMC-derived immune cell types. A) Preliminary gating for single lyphocytes. B) Example staining for CD122 (red), the corresponding isotype control (blue), and unstained cells (black). C) Gating for live T helper and T regulatory cells during receptor quantification. D) Live cell NK cell gating. E) Live cell CD8+ T cell gating. F) Gating for fixed T helper and T regulatory cells during pSTAT5 quantification. G) Fixed CD8+ T cell and NK cell gating.

    Techniques Used: Derivative Assay, Staining

    Model accurately predicts cell type-specific response across a panel of PBMC-derived cell types. A) Comparison of two replicates measuring pSTAT5 response to a dose-response of IL-2/-15, time course, and panel of PBMC-derived cell types. B) Both experimentally-derived and model-predicted EC 50 s of dose response across IL-2/-15 and all 10 cell types. EC 50 s are shown for 1 hr time point. C) Pearson correlation coefficients between model prediction and experimental measurements for all 10 cell populations (full data shown in Fig. S5 ). D–I) pSTAT5 response to IL-2 (D-F) or IL-15 (G-I) dose responses in NK, CD8+, and T reg cells.
    Figure Legend Snippet: Model accurately predicts cell type-specific response across a panel of PBMC-derived cell types. A) Comparison of two replicates measuring pSTAT5 response to a dose-response of IL-2/-15, time course, and panel of PBMC-derived cell types. B) Both experimentally-derived and model-predicted EC 50 s of dose response across IL-2/-15 and all 10 cell types. EC 50 s are shown for 1 hr time point. C) Pearson correlation coefficients between model prediction and experimental measurements for all 10 cell populations (full data shown in Fig. S5 ). D–I) pSTAT5 response to IL-2 (D-F) or IL-15 (G-I) dose responses in NK, CD8+, and T reg cells.

    Techniques Used: Derivative Assay

    A reaction model captures cytokine-cytokine interactions. A) Schematic of IL-4 and IL-7 receptor complexes competing for γ c and generating distinct pSTAT signals. B-C) Fitting model to experimental data. Experimental measurements are denoted by triangles. Shaded areas represent the 25-75% and 10-90% confidence intervals of model predictions. pSTAT5 and pSTAT6 were measured for IL-7 and IL-4 experiments, respectively. B) Singlecytokine pSTAT dose-response measurements for 10 min of exposure to IL-4 and IL-7. C) Percent inhibition of the second cytokine’s pSTAT response in a dual-cytokine dose-response experiment. Human PBMC-derived T cells (CD4 + TCR + CCR7 high ) were pretreated with various concentrations of one cytokine for 10 min before being stimulated with a fixed concentration (50 pg/mL IL-7 or 100 pg/mL IL-4) of the other cytokine for an additional 10 min. D) Model predictions for percent inhibition of the second cytokine’s pSTAT response in a dual-cytokine dose-response experiment with the assumption that active species are endocytosed at the same rate as inactive species (k endo,a = k endo ). E) Model predictions for percent of γ c on the cell surface when exposed to 100 pg/mL of either IL-7 or IL-4 for 100 min. F) Violin plot of K a values obtained via posterior distributions of k fwd / k rev for krev parameters corresponding to different complexes competing for the common γ c ( Fig. 1A ). G–I) Posterior distributions from fitting to data. Scaling constants C 5 and C 6 have units of # × cell −1 , k fwd has units of cell × # −1 × min −1 , and f sort is unitless
    Figure Legend Snippet: A reaction model captures cytokine-cytokine interactions. A) Schematic of IL-4 and IL-7 receptor complexes competing for γ c and generating distinct pSTAT signals. B-C) Fitting model to experimental data. Experimental measurements are denoted by triangles. Shaded areas represent the 25-75% and 10-90% confidence intervals of model predictions. pSTAT5 and pSTAT6 were measured for IL-7 and IL-4 experiments, respectively. B) Singlecytokine pSTAT dose-response measurements for 10 min of exposure to IL-4 and IL-7. C) Percent inhibition of the second cytokine’s pSTAT response in a dual-cytokine dose-response experiment. Human PBMC-derived T cells (CD4 + TCR + CCR7 high ) were pretreated with various concentrations of one cytokine for 10 min before being stimulated with a fixed concentration (50 pg/mL IL-7 or 100 pg/mL IL-4) of the other cytokine for an additional 10 min. D) Model predictions for percent inhibition of the second cytokine’s pSTAT response in a dual-cytokine dose-response experiment with the assumption that active species are endocytosed at the same rate as inactive species (k endo,a = k endo ). E) Model predictions for percent of γ c on the cell surface when exposed to 100 pg/mL of either IL-7 or IL-4 for 100 min. F) Violin plot of K a values obtained via posterior distributions of k fwd / k rev for krev parameters corresponding to different complexes competing for the common γ c ( Fig. 1A ). G–I) Posterior distributions from fitting to data. Scaling constants C 5 and C 6 have units of # × cell −1 , k fwd has units of cell × # −1 × min −1 , and f sort is unitless

    Techniques Used: Inhibition, Derivative Assay, Concentration Assay

    3) Product Images from "BET protein targeting suppresses the PD-1/PD-L1 pathway in triple-negative breast cancer and elicits anti-tumor immune response"

    Article Title: BET protein targeting suppresses the PD-1/PD-L1 pathway in triple-negative breast cancer and elicits anti-tumor immune response

    Journal: Cancer letters

    doi: 10.1016/j.canlet.2019.08.013

    BET proteins regulate effector T celli interferon-γ secretion to induce PD-L1 expression in triple-negative breast cells.(A) Conditioned media generated from cultures of either unstimulated or activated PBMCs in presence of control or active JQ1 (400nM) for 3 days were profiled for cytokine/chemokine expression by multiplex analysis. Heatmap shows the Z score hierarchical clustering analysis for each analyte. Five clusters identified by letters were identified. The number of significantly altered genes in each cluster is indicated. (B) Expression of Cluster B analytes is detailed. Each dot represents an experiment. (C) To validate the central role of IFNγ, MDA-MB-231cells were co-cultured as previously described with unstimulated PBMCs or activated PBMCs under regular conditions or in the presence of a neutralizing anti-IFNγ antibody for 24h prior to evaluation of PD-L1 expression by flow cytometry. Histograms represent three independent experiments. Errors bars represent SEM of three independent experiments. ns, p > 0.05; *, p
    Figure Legend Snippet: BET proteins regulate effector T celli interferon-γ secretion to induce PD-L1 expression in triple-negative breast cells.(A) Conditioned media generated from cultures of either unstimulated or activated PBMCs in presence of control or active JQ1 (400nM) for 3 days were profiled for cytokine/chemokine expression by multiplex analysis. Heatmap shows the Z score hierarchical clustering analysis for each analyte. Five clusters identified by letters were identified. The number of significantly altered genes in each cluster is indicated. (B) Expression of Cluster B analytes is detailed. Each dot represents an experiment. (C) To validate the central role of IFNγ, MDA-MB-231cells were co-cultured as previously described with unstimulated PBMCs or activated PBMCs under regular conditions or in the presence of a neutralizing anti-IFNγ antibody for 24h prior to evaluation of PD-L1 expression by flow cytometry. Histograms represent three independent experiments. Errors bars represent SEM of three independent experiments. ns, p > 0.05; *, p

    Techniques Used: Expressing, Generated, Multiplex Assay, Multiple Displacement Amplification, Cell Culture, Flow Cytometry

    BET protein inhibition reduces PD-1 expression in activated T cells. (A) Flow cytometry experimental design and gating strategy to analyze PD-1 expression in CD4 + and CD8 + T cells. These experiments were repeated four times. At least 1 × 10 6 viable total T cells (CD3 + ) were counted and PD-1 + proportions were determined among the CD4 + and CD8 + T cell populations. (B) 1 × 10 7 ) and flow cytometry analysis. (C) Unstimulated PBMCs or activated T cells were cultured as described above with CFSE to determine their proliferation in presence of control or active JQ1 (400 nM) for 3 days. The numbers above the peaks indicate the numbers of cell divisions. The histograms indicate the quantitation of three independent experiments. (D) CD69 + , CD25 + and CD137 + percentages were determined among the CD4 + and CD8 + T cell populations. (E) Unstimulated or activated PBMCs from 20 female patients were analyzed by qRT-PCR for PDCD1 upon 400 nM JQ1 treatment for 24h. (F) Activated PBMCs from 20 female patients were analyzed by qRT-PCR for PDCD1, BRD2 , BRD3 and BRD4 expression. Spearman’s correlation was calculated for each pair of plotted genes and indicated in the corresponding plots along with the calculated p -values. Errors bars represent SEM of four independent experiments. ns, p > 0.05; *, p
    Figure Legend Snippet: BET protein inhibition reduces PD-1 expression in activated T cells. (A) Flow cytometry experimental design and gating strategy to analyze PD-1 expression in CD4 + and CD8 + T cells. These experiments were repeated four times. At least 1 × 10 6 viable total T cells (CD3 + ) were counted and PD-1 + proportions were determined among the CD4 + and CD8 + T cell populations. (B) 1 × 10 7 ) and flow cytometry analysis. (C) Unstimulated PBMCs or activated T cells were cultured as described above with CFSE to determine their proliferation in presence of control or active JQ1 (400 nM) for 3 days. The numbers above the peaks indicate the numbers of cell divisions. The histograms indicate the quantitation of three independent experiments. (D) CD69 + , CD25 + and CD137 + percentages were determined among the CD4 + and CD8 + T cell populations. (E) Unstimulated or activated PBMCs from 20 female patients were analyzed by qRT-PCR for PDCD1 upon 400 nM JQ1 treatment for 24h. (F) Activated PBMCs from 20 female patients were analyzed by qRT-PCR for PDCD1, BRD2 , BRD3 and BRD4 expression. Spearman’s correlation was calculated for each pair of plotted genes and indicated in the corresponding plots along with the calculated p -values. Errors bars represent SEM of four independent experiments. ns, p > 0.05; *, p

    Techniques Used: Inhibition, Expressing, Flow Cytometry, Cell Culture, Quantitation Assay, Quantitative RT-PCR

    BET protein inhibition reduces the PD-1/PD-L1 axis in a T cell/triple-negative breast cell co-culture system. (A) Diagram of the co-culture experimental design. These experiments have been repeated three times. Atleast 1×10 5 viable tumor cell sand1×10 6 T cells were analysed by flow cytometry. (B–D) MDA-MB-231 cells alone (N.C.) or co-cultured (Coc.) with unstimulated (Unstim.) PBMCs or activated T cells obtained from human normal blood donors at a ratio of 1:10 (tumor cells: T cells) were analyzed forPD-L1 expression (B). The histograms represent the quantification of the illustrated flow cytometry curves. (C–D) PD-1 + proportions were determined among theCD4 + (C) andCD8 + T cell (D) populations. (E) Diagram of the conditioned media generation and subsequent experimental design. These experiments have been repeated three times. (F)MDA-MB-231 and SUM149PT cells were cultured with the generated conditioned media from (E) for3 days prior to PD-L1 expression analysis. The histograms represent the quantification of the illustrated flow cytometry curves. Errors bars represent SEM of four independent experiments. ns, p > 0.05; *, p
    Figure Legend Snippet: BET protein inhibition reduces the PD-1/PD-L1 axis in a T cell/triple-negative breast cell co-culture system. (A) Diagram of the co-culture experimental design. These experiments have been repeated three times. Atleast 1×10 5 viable tumor cell sand1×10 6 T cells were analysed by flow cytometry. (B–D) MDA-MB-231 cells alone (N.C.) or co-cultured (Coc.) with unstimulated (Unstim.) PBMCs or activated T cells obtained from human normal blood donors at a ratio of 1:10 (tumor cells: T cells) were analyzed forPD-L1 expression (B). The histograms represent the quantification of the illustrated flow cytometry curves. (C–D) PD-1 + proportions were determined among theCD4 + (C) andCD8 + T cell (D) populations. (E) Diagram of the conditioned media generation and subsequent experimental design. These experiments have been repeated three times. (F)MDA-MB-231 and SUM149PT cells were cultured with the generated conditioned media from (E) for3 days prior to PD-L1 expression analysis. The histograms represent the quantification of the illustrated flow cytometry curves. Errors bars represent SEM of four independent experiments. ns, p > 0.05; *, p

    Techniques Used: Inhibition, Co-Culture Assay, Flow Cytometry, Multiple Displacement Amplification, Cell Culture, Expressing, Generated

    4) Product Images from "The HIV-1 Env gp120 Inner Domain Shapes the Phe43 Cavity and the CD4 Binding Site"

    Article Title: The HIV-1 Env gp120 Inner Domain Shapes the Phe43 Cavity and the CD4 Binding Site

    Journal: mBio

    doi: 10.1128/mBio.00280-20

    Primary viruses harboring a T375 residue are highly susceptible to ADCC responses mediated by HIV + sera in the presence of CD4mc. Primary CD4 + T cells were infected with clade B primary HIV-1 and their variants (A to D) or with a panel of TF and chronic viruses from clades B and C (E to H). At 48 h postinfection, cell surface staining was performed with 10 different HIV + sera. The graphs shown represent (A and E) the compiled mean fluorescence intensities (MFI) obtained with 10 HIV + sera and (B and F) the fold increase in MFI in the presence of CD4mc calculated on the infected (p24 + ) population. Infected primary CD4 + T cells were also used as target cells, and autologous PBMCs were used as effector cells in a FACS-based ADCC assay. The graphs shown represent (C and G) the ADCC values and (D and H) the increases (in percentage points) in the levels of ADCC obtained in the presence of CD4mc with 10 HIV + sera. These results were obtained in at least 2 independent experiments. Error bars indicate means ± SEM. Statistical significance was tested using a paired t test or a Wilcoxon signed-rank test (A to E and G) or an unpaired t test or Mann-Whitney U test (F and H), based on statistical normality (*, P
    Figure Legend Snippet: Primary viruses harboring a T375 residue are highly susceptible to ADCC responses mediated by HIV + sera in the presence of CD4mc. Primary CD4 + T cells were infected with clade B primary HIV-1 and their variants (A to D) or with a panel of TF and chronic viruses from clades B and C (E to H). At 48 h postinfection, cell surface staining was performed with 10 different HIV + sera. The graphs shown represent (A and E) the compiled mean fluorescence intensities (MFI) obtained with 10 HIV + sera and (B and F) the fold increase in MFI in the presence of CD4mc calculated on the infected (p24 + ) population. Infected primary CD4 + T cells were also used as target cells, and autologous PBMCs were used as effector cells in a FACS-based ADCC assay. The graphs shown represent (C and G) the ADCC values and (D and H) the increases (in percentage points) in the levels of ADCC obtained in the presence of CD4mc with 10 HIV + sera. These results were obtained in at least 2 independent experiments. Error bars indicate means ± SEM. Statistical significance was tested using a paired t test or a Wilcoxon signed-rank test (A to E and G) or an unpaired t test or Mann-Whitney U test (F and H), based on statistical normality (*, P

    Techniques Used: Infection, Staining, Fluorescence, FACS, ADCC Assay, MANN-WHITNEY

    Phe43 cavity and inner domain changes enhance susceptibility of the CRF01_AE HIV-1 strain to ADCC responses mediated by HIV + sera in the presence of CD4mc. (A to C) Cell surface staining of primary CD4 + T cells infected with a CRF01_AE transmitted-founder (TF) strain (40061) and variants with 10 different HIV + sera. (A) Histograms depicting representative HIV + serum staining. (B and C) The graphs shown represent the compiled mean fluorescence intensities (MFI) obtained with 10 HIV + sera (B) and the fold increase in MFI in the presence of CD4mc calculated for the infected (p24 + ) population (C). (D and E) Infected primary CD4 + T cells were also used as target cells, and autologous PBMCs were used as effector cells in a FACS-based ADCC assay. The graphs shown represent (D) the ADCC values and (E) the increase (in percentage points) of ADCC levels obtained in the presence of CD4mc with 10 HIV + sera. These results were obtained in at least 2 independent experiments. Error bars indicate means ± SEM. Statistical significance was tested using a paired t test or a Wilcoxon rank test based on statistical normality (**, P
    Figure Legend Snippet: Phe43 cavity and inner domain changes enhance susceptibility of the CRF01_AE HIV-1 strain to ADCC responses mediated by HIV + sera in the presence of CD4mc. (A to C) Cell surface staining of primary CD4 + T cells infected with a CRF01_AE transmitted-founder (TF) strain (40061) and variants with 10 different HIV + sera. (A) Histograms depicting representative HIV + serum staining. (B and C) The graphs shown represent the compiled mean fluorescence intensities (MFI) obtained with 10 HIV + sera (B) and the fold increase in MFI in the presence of CD4mc calculated for the infected (p24 + ) population (C). (D and E) Infected primary CD4 + T cells were also used as target cells, and autologous PBMCs were used as effector cells in a FACS-based ADCC assay. The graphs shown represent (D) the ADCC values and (E) the increase (in percentage points) of ADCC levels obtained in the presence of CD4mc with 10 HIV + sera. These results were obtained in at least 2 independent experiments. Error bars indicate means ± SEM. Statistical significance was tested using a paired t test or a Wilcoxon rank test based on statistical normality (**, P

    Techniques Used: Staining, Infection, Fluorescence, FACS, ADCC Assay

    5) Product Images from "A New Family of Small-Molecule CD4-Mimetic Compounds Contacts Highly Conserved Aspartic Acid 368 of HIV-1 gp120 and Mediates Antibody-Dependent Cellular Cytotoxicity"

    Article Title: A New Family of Small-Molecule CD4-Mimetic Compounds Contacts Highly Conserved Aspartic Acid 368 of HIV-1 gp120 and Mediates Antibody-Dependent Cellular Cytotoxicity

    Journal: Journal of Virology

    doi: 10.1128/JVI.01325-19

    ( S )-MCG-IV-210 blocks uninfected bystander ADCC killing. Uninfected primary CD4 + T cells were stained with the cellular dye eFluor-450 and cocultured with unstained NL4-3.ADA.GFP WT virus-infected autologous cells for 72 h. (A) The ability of HIV + plasma to recognize uninfected bystander cells in the presence of 50 μM (+)-BNM-III-170 or ( S )-MCG-IV-210 was evaluated by FACS analysis. (B) These uninfected eFluor-450 + cells were also used as target cells for ADCC with autologous PBMCs and 5 HIV + plasma samples in the presence of 50 μM (+)-BNM-III-170 or ( S )-MCG-IV-210. Statistical significance was evaluated using a paired t test (*, P
    Figure Legend Snippet: ( S )-MCG-IV-210 blocks uninfected bystander ADCC killing. Uninfected primary CD4 + T cells were stained with the cellular dye eFluor-450 and cocultured with unstained NL4-3.ADA.GFP WT virus-infected autologous cells for 72 h. (A) The ability of HIV + plasma to recognize uninfected bystander cells in the presence of 50 μM (+)-BNM-III-170 or ( S )-MCG-IV-210 was evaluated by FACS analysis. (B) These uninfected eFluor-450 + cells were also used as target cells for ADCC with autologous PBMCs and 5 HIV + plasma samples in the presence of 50 μM (+)-BNM-III-170 or ( S )-MCG-IV-210. Statistical significance was evaluated using a paired t test (*, P

    Techniques Used: Staining, Infection, FACS

    ( S )-MCG-IV-210 stabilizes state 2A at the surface of infected cells and virions. (A to D) For cell surface staining with A32-AF647, primary CD4 T cells isolated from PBMCs were infected with HIV-1 CH58TF for 48 h. Cells were then incubated with A32-AF647 together with 5 μg/ml 17b (A to C) or 1:1,000-diluted HIV + plasma from infected individuals (D) in the presence of DMSO, 50 μM (+)-BNM-III-170, or 50 μM ( S )-MCG-IV-210 at 37°C. The mean fluorescence intensity (MFI) of A32-AF647 was measured by flow cytometry. (E) For the VCA, virus produced from HEK293T cells cotransfected with plasmids pNL4.3 Luc Env − , HIV-1 CH58TF , and VSV-G was incubated with or without 5 μg/ml 17b in the presence of DMSO, 50 μM (+)-BNM-III-170, or 50 μM ( S )-MCG-IV-210 at 37°C for 1 h. Virus was then applied to ELISA plates coated with Ab 2G12, 17b, A32, or C11 overnight at 4°C. Free virions were washed away, and HEK293T cells were added to the well. After 48 h, cells were lysed, and luciferase activity was measured. To compare the binding capacities of different Abs, the relative ratio of the luciferase activity to the luciferase activity of 2G12 was calculated. Data shown are the means ± SD from at least three independent experiments. Statistical significance was evaluated using a Mann-Whitney unpaired t test (C) or a Wilcoxon paired t test (D) (*, P
    Figure Legend Snippet: ( S )-MCG-IV-210 stabilizes state 2A at the surface of infected cells and virions. (A to D) For cell surface staining with A32-AF647, primary CD4 T cells isolated from PBMCs were infected with HIV-1 CH58TF for 48 h. Cells were then incubated with A32-AF647 together with 5 μg/ml 17b (A to C) or 1:1,000-diluted HIV + plasma from infected individuals (D) in the presence of DMSO, 50 μM (+)-BNM-III-170, or 50 μM ( S )-MCG-IV-210 at 37°C. The mean fluorescence intensity (MFI) of A32-AF647 was measured by flow cytometry. (E) For the VCA, virus produced from HEK293T cells cotransfected with plasmids pNL4.3 Luc Env − , HIV-1 CH58TF , and VSV-G was incubated with or without 5 μg/ml 17b in the presence of DMSO, 50 μM (+)-BNM-III-170, or 50 μM ( S )-MCG-IV-210 at 37°C for 1 h. Virus was then applied to ELISA plates coated with Ab 2G12, 17b, A32, or C11 overnight at 4°C. Free virions were washed away, and HEK293T cells were added to the well. After 48 h, cells were lysed, and luciferase activity was measured. To compare the binding capacities of different Abs, the relative ratio of the luciferase activity to the luciferase activity of 2G12 was calculated. Data shown are the means ± SD from at least three independent experiments. Statistical significance was evaluated using a Mann-Whitney unpaired t test (C) or a Wilcoxon paired t test (D) (*, P

    Techniques Used: Infection, Staining, Isolation, Incubation, Fluorescence, Flow Cytometry, Produced, Enzyme-linked Immunosorbent Assay, Luciferase, Activity Assay, Binding Assay, MANN-WHITNEY

    MCG analogs sensitize HIV-1-infected cells to ADCC and viral particles to neutralization by nnAbs. (A and B) Primary CD4 T cells isolated from PBMCs were infected with HIV-1 CH58TF for 48 h. For cell surface staining, 5 μg/ml 17b (A) or 1:1,000-diluted HIV + plasma ( n = 15) (B) was used in the presence of the different MCG analogs (50 μM), (+)-BNM-III-170, or an equivalent volume of the vehicle (DMSO). An Alexa Fluor 647-conjugated anti-human IgG secondary Ab was then used for fluorescence labeling. (C) For ADCC, infected cells were used as target cells in a FACS-based ADCC assay that measures the killing of infected (p24 + ). The assay determines susceptibility to ADCC mediated by a 1/1,000 dilution of plasma from 15 HIV-1-infected individuals in the presence of the different MCG analogs (50 μM), (+)-BNM-III-170 (50 μM), or an equivalent volume of the vehicle (DMSO). (D) The correlation between cell surface staining with HIV + plasma and ADCC was calculated using the Spearman rank correlation. (E) To evaluate the direct virus-neutralizing ability of the analogs, HIV-1 CH58TF was incubated with the indicated amounts of different compounds or DMSO for 1 h at 37°C and then added to TZM-bl cells. After incubation for 48 h at 37°C, luciferase activity was measured. Relative infectivity was calculated as the percentage of the value seen in the absence of the compound. (F) To measure the ability of ( S )-MCG-IV-210 to sensitize viral particles to neutralization by otherwise nonneutralizing Ab 17b, HIV-1 CH58TF was incubated with the indicated amounts of 17b in the presence of 0.5 μM (+)-BNM-III-170, 20 μM ( S )-MCG-IV-210, or DMSO for 1 h at 37°C and then added to TZM-bl cells. After incubation for 48 h at 37°C, luciferase activity was measured. Relative infectivity was calculated as the percentage of the value seen in the absence of the compound. Data shown are the means ± SD from at least three independent experiments. Statistical significance was evaluated using a Mann-Whitney unpaired t test (A) or a Wilcoxon paired t test (B and C) (**, P
    Figure Legend Snippet: MCG analogs sensitize HIV-1-infected cells to ADCC and viral particles to neutralization by nnAbs. (A and B) Primary CD4 T cells isolated from PBMCs were infected with HIV-1 CH58TF for 48 h. For cell surface staining, 5 μg/ml 17b (A) or 1:1,000-diluted HIV + plasma ( n = 15) (B) was used in the presence of the different MCG analogs (50 μM), (+)-BNM-III-170, or an equivalent volume of the vehicle (DMSO). An Alexa Fluor 647-conjugated anti-human IgG secondary Ab was then used for fluorescence labeling. (C) For ADCC, infected cells were used as target cells in a FACS-based ADCC assay that measures the killing of infected (p24 + ). The assay determines susceptibility to ADCC mediated by a 1/1,000 dilution of plasma from 15 HIV-1-infected individuals in the presence of the different MCG analogs (50 μM), (+)-BNM-III-170 (50 μM), or an equivalent volume of the vehicle (DMSO). (D) The correlation between cell surface staining with HIV + plasma and ADCC was calculated using the Spearman rank correlation. (E) To evaluate the direct virus-neutralizing ability of the analogs, HIV-1 CH58TF was incubated with the indicated amounts of different compounds or DMSO for 1 h at 37°C and then added to TZM-bl cells. After incubation for 48 h at 37°C, luciferase activity was measured. Relative infectivity was calculated as the percentage of the value seen in the absence of the compound. (F) To measure the ability of ( S )-MCG-IV-210 to sensitize viral particles to neutralization by otherwise nonneutralizing Ab 17b, HIV-1 CH58TF was incubated with the indicated amounts of 17b in the presence of 0.5 μM (+)-BNM-III-170, 20 μM ( S )-MCG-IV-210, or DMSO for 1 h at 37°C and then added to TZM-bl cells. After incubation for 48 h at 37°C, luciferase activity was measured. Relative infectivity was calculated as the percentage of the value seen in the absence of the compound. Data shown are the means ± SD from at least three independent experiments. Statistical significance was evaluated using a Mann-Whitney unpaired t test (A) or a Wilcoxon paired t test (B and C) (**, P

    Techniques Used: Infection, Neutralization, Isolation, Staining, Fluorescence, Labeling, FACS, ADCC Assay, Incubation, Luciferase, Activity Assay, MANN-WHITNEY

    6) Product Images from "Antimetabolite pemetrexed primes a favorable tumor microenvironment for immune checkpoint blockade therapy"

    Article Title: Antimetabolite pemetrexed primes a favorable tumor microenvironment for immune checkpoint blockade therapy

    Journal: Journal for Immunotherapy of Cancer

    doi: 10.1136/jitc-2020-001392

    Knockdown of thymidylate synthase (TS) induces programmed death-ligand 1 (PD-L1) expression in non-small-cell lung cancer (NSCLC) cells and decreases the production of interleukin-2 (IL-2) by activated T cells in the NSCLC and T cell coculture system. (A–D), CL1-5 or CL141 cells transfected with control-siRNA (siCtrl) or TS -siRNA (siTS) oligonucleotides were lysed and analyzed by qRT-PCR (A), flow cytometry (B), or immunoblotting (C, D) 72 hours after transfection. Data are shown as means and SD for three independent experiments (n=3). (E, F) CL1-5 or CL141 cells transfected with control-siRNA (siCtrl), TS-siRNA (siTS) and RelA-siRNA (siRelA) oligonucleotides in different combination were lysed and analyzed by qRT-PCR (E) or immunoblotting (F), 72 hours after transfection. (G, H) CL1-5 or CL141 cells were transfected with siCtrl or siTS siRNA oligonucleotides for 24 hours and followed by cocultured with Jurkat T-cells or PBMCs at different cancer to T cell ratios in the presence of the 1×T cell stimulation cocktail for additional 48 hours. (G) IL-2 levels were measured by ELISA. (H) The levels of CD69 and intracellular IL-2 produced by Jurkat T-cells or PBMCs were measured by flow cytometry. **P
    Figure Legend Snippet: Knockdown of thymidylate synthase (TS) induces programmed death-ligand 1 (PD-L1) expression in non-small-cell lung cancer (NSCLC) cells and decreases the production of interleukin-2 (IL-2) by activated T cells in the NSCLC and T cell coculture system. (A–D), CL1-5 or CL141 cells transfected with control-siRNA (siCtrl) or TS -siRNA (siTS) oligonucleotides were lysed and analyzed by qRT-PCR (A), flow cytometry (B), or immunoblotting (C, D) 72 hours after transfection. Data are shown as means and SD for three independent experiments (n=3). (E, F) CL1-5 or CL141 cells transfected with control-siRNA (siCtrl), TS-siRNA (siTS) and RelA-siRNA (siRelA) oligonucleotides in different combination were lysed and analyzed by qRT-PCR (E) or immunoblotting (F), 72 hours after transfection. (G, H) CL1-5 or CL141 cells were transfected with siCtrl or siTS siRNA oligonucleotides for 24 hours and followed by cocultured with Jurkat T-cells or PBMCs at different cancer to T cell ratios in the presence of the 1×T cell stimulation cocktail for additional 48 hours. (G) IL-2 levels were measured by ELISA. (H) The levels of CD69 and intracellular IL-2 produced by Jurkat T-cells or PBMCs were measured by flow cytometry. **P

    Techniques Used: Expressing, Transfection, Quantitative RT-PCR, Flow Cytometry, Cell Stimulation, Enzyme-linked Immunosorbent Assay, Produced

    Pemetrexed (PEM) and 5-fluorouracil (5-FU) suppress the production of interleukin-2 (IL-2) and interferon (IFN)-γ by activated T cells in the non-small-cell lung cancer (NSCLC) and T cell coculture system. (A–D) CL1-5 or CL141 cells were preincubated with 100 nM PEM, 5 µM 5-FU or the vehicle control (PBS) for 48 hours, and subsequently cocultured with activated Jurkat T-cells (A, B) or PBMCs (C, D) at different cancer to T cell ratios in the presence of 1× T cell stimulation cocktail for additional 48 hours. The levels of IL-2 and IFN-γ were measured by ELISA. Data are shown as means and SD for three independent experiments (n=3). (E–G) CL1-5 or CL141 cells were preincubated with 100 nM PEM, 5 µM 5-FU or the vehicle control (PBS) for 48 hours, and subsequently cocultured with activated Jurkat T-cells in the presence of 1× T cell stimulation cocktail for additional 48 hours. The levels of CD69 and intracecllular IL-2 produced by Jurkat T-cells were measured by flow cytometry. (E) Representative dot plots for the indicated cell percentages determined by flow cytometry. (F, G) Quantitative plots for the CD69 and intracellular IL-2 staining in CD45 + T-cells. (H) T-cell-meditated killing of PD-L1-expressing CL141 NSCLC cells. CL141 cells stably expressing nuclear RFP protein were pretreated with or without 50 nM PEM for 48 hours and cocultured with activated Jurkat T-cells with or without 10 µg/mL of anti-PD-L1 antibody for additional 48 hours. Relative cell viability was measured by RFP signaling after 48 hours of coincubation and the results were normalized at the zero-time point. Data are shown as means and s.e.m. for three independent experiments (n=3). *P
    Figure Legend Snippet: Pemetrexed (PEM) and 5-fluorouracil (5-FU) suppress the production of interleukin-2 (IL-2) and interferon (IFN)-γ by activated T cells in the non-small-cell lung cancer (NSCLC) and T cell coculture system. (A–D) CL1-5 or CL141 cells were preincubated with 100 nM PEM, 5 µM 5-FU or the vehicle control (PBS) for 48 hours, and subsequently cocultured with activated Jurkat T-cells (A, B) or PBMCs (C, D) at different cancer to T cell ratios in the presence of 1× T cell stimulation cocktail for additional 48 hours. The levels of IL-2 and IFN-γ were measured by ELISA. Data are shown as means and SD for three independent experiments (n=3). (E–G) CL1-5 or CL141 cells were preincubated with 100 nM PEM, 5 µM 5-FU or the vehicle control (PBS) for 48 hours, and subsequently cocultured with activated Jurkat T-cells in the presence of 1× T cell stimulation cocktail for additional 48 hours. The levels of CD69 and intracecllular IL-2 produced by Jurkat T-cells were measured by flow cytometry. (E) Representative dot plots for the indicated cell percentages determined by flow cytometry. (F, G) Quantitative plots for the CD69 and intracellular IL-2 staining in CD45 + T-cells. (H) T-cell-meditated killing of PD-L1-expressing CL141 NSCLC cells. CL141 cells stably expressing nuclear RFP protein were pretreated with or without 50 nM PEM for 48 hours and cocultured with activated Jurkat T-cells with or without 10 µg/mL of anti-PD-L1 antibody for additional 48 hours. Relative cell viability was measured by RFP signaling after 48 hours of coincubation and the results were normalized at the zero-time point. Data are shown as means and s.e.m. for three independent experiments (n=3). *P

    Techniques Used: Cell Stimulation, Enzyme-linked Immunosorbent Assay, Produced, Flow Cytometry, Staining, Expressing, Stable Transfection

    Related Articles

    Cell Culture:

    Article Title: DNA-PKcs controls calcineurin mediated IL-2 production in T lymphocytes
    Article Snippet: Dynabeads Human T-Activator CD3/CD28 was purchased from Thermo Fisherscientific (Waltham, MA). .. Cell culture Human peripheral blood mononuclear cells (PBMC,) and Jurkat cells were purchased from ATCC (PCS-800-011, Manassas, VA). ..

    Article Title: Tendency of K562 Chronic Myeloid Leukemia Cells Towards Cell Reprogramming
    Article Snippet: .. Cell Culture Human CML cell line K562 and human peripheral blood mononuclear cells (PBMCs) were obtained from ATCC and Lonza, respectively. .. Cells were cultured in RPMI 1640 supplemented with 10% fetal bovine serum, 50 U/mL penicillin, 50 µg/mL streptomycin, and 1% L‐glutamine at 37 °C in 5% CO2 .

    Article Title: Nucleotide ecto-enzyme metabolic pattern and spatial distribution in calcific aortic valve disease; its relation to pathological changes and clinical presentation
    Article Snippet: .. Isolated human peripheral blood mononuclear cells and monocyte/macrophage cells (SC line, ATCC, cat. CRL-9855) that were used at passage 4, were plated in 24-well cell culture plate at a density 0.2 × 106 per well in a total volume of 1 mL HBSS. .. Mice aortic roots were cleaned of surrounding tissues as described above and used for experiment.

    Gradient Centrifugation:

    Article Title: Critical Role for the NLRP3 Inflammasome in Mediating IL-1β Production in Shigella sonnei-Infected Macrophages
    Article Snippet: Cell Lines and Culture Mouse J774A.1 macrophages and human THP-1 monocytes were purchased from the American Type Culture Collection (Rockville, MD). .. THP-1 macrophages were differentiated from THP-1 monocytes by treatment with 50 nM PMA for 48 h. Human peripheral blood mononuclear cells (PBMCs) were separated from whole blood from healthy volunteers by density gradient centrifugation using Histopaque-1077 , and all experimental protocols were performed in accordance with the guidelines and regulations provided and accepted by the Institutional Review Board of the Tri-Service General Hospital, National Defense Medical Center and the volunteers' informed consent (TSGH-IRB-2-106-05-190 and TSGH-IRB-2-106-05-009). .. Mouse primary bone marrow derived macrophages (BMDM) were prepared from bone marrow collected from C57BL/6 mouse femur and tibia by differentiating in the M-CSF containing medium for 7 days.

    Labeling:

    Article Title: Nkx2‐5 Is Expressed in Atherosclerotic Plaques and Attenuates Development of Atherosclerosis in Apolipoprotein E–Deficient Mice
    Article Snippet: Cells were infected with Ad‐Nkx2‐5 or Ad‐EV, followed by treating with tumor necrosis factor alpha (TNFα; 10 ng/mL) for 6 hours. .. Fresh adult human PBMCs (PCS‐800‐011; ATCC) were labeled with calcein acetomethoxy (AM) dye: PBMCs were pelleted at 240g for 10 minutes, resuspended in 1 mL of culture medium with 2.5 μmol/L of calcein AM from the kit, and incubated at 37°C (5% CO2 ) for 30 minutes. .. PBMCs were then washed 3 times with HAEC media and added to HAEC cells (150 000 labeled PBMCs per chamber).

    Incubation:

    Article Title: Nkx2‐5 Is Expressed in Atherosclerotic Plaques and Attenuates Development of Atherosclerosis in Apolipoprotein E–Deficient Mice
    Article Snippet: Cells were infected with Ad‐Nkx2‐5 or Ad‐EV, followed by treating with tumor necrosis factor alpha (TNFα; 10 ng/mL) for 6 hours. .. Fresh adult human PBMCs (PCS‐800‐011; ATCC) were labeled with calcein acetomethoxy (AM) dye: PBMCs were pelleted at 240g for 10 minutes, resuspended in 1 mL of culture medium with 2.5 μmol/L of calcein AM from the kit, and incubated at 37°C (5% CO2 ) for 30 minutes. .. PBMCs were then washed 3 times with HAEC media and added to HAEC cells (150 000 labeled PBMCs per chamber).

    Activation Assay:

    Article Title: Protein Kinase D1: A New Component in Toll-Like Receptor 9 Signaling
    Article Snippet: Our results indicate that certain pathogenic DNA, such as CpG motif containing DNA, induces activation of one or more PKD family members in macrophages. .. Because, in addition to macrophages, other types of cells, including B cells and DCs, also express TLR9 (the CpG motif containing DNA receptor) and respond to CpG motif containing DNA, we further investigated whether CpG motif containing DNA can induce activation of PKD family members in B cells, pDCs, cDCs, and human PBMCs. ..

    other:

    Article Title: Potency of Combining Eucalyptus camaldulensis subsp. camaldulensis with Low-Dose Cisplatin in A549 Human Lung Adenocarcinomas and MCF-7 Breast Adenocarcinoma
    Article Snippet: Peripheral Blood Mononuclear Cells (PBMC)Human peripheral blood mononuclear cells (PBMCs) obtained from ATCC, Manassas, VA, USA, comprising lymphocytes (B-cells, T-cells, and NK-cells), monocytes, and dendritic cells, were frequently used for the evaluation of immune responses.

    Isolation:

    Article Title: Nucleotide ecto-enzyme metabolic pattern and spatial distribution in calcific aortic valve disease; its relation to pathological changes and clinical presentation
    Article Snippet: .. Isolated human peripheral blood mononuclear cells and monocyte/macrophage cells (SC line, ATCC, cat. CRL-9855) that were used at passage 4, were plated in 24-well cell culture plate at a density 0.2 × 106 per well in a total volume of 1 mL HBSS. .. Mice aortic roots were cleaned of surrounding tissues as described above and used for experiment.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    ATCC human peripheral blood mononuclear cells
    Stenotic aortic valve immune infiltrate is a smaller source of nucleotide-degrading ecto-nucleotidases but a larger of adenosine deaminase. Simplified protocol of stenotic aortic valve cell isolation, including endothelial <t>cells</t> (first step of isolation, vWF positive or CD31 high positive), interstitial cells (second step of isolation; Vimentin positive, Vim+) and immune cells (first and second step of isolation; CD45 positive, CD45+) ( a ). Flow cytometry analysis of CD45 positive cells (immune cells) as a percentage of total isolated cells after first step of isolation (cells located in the upper layers of the valve) and second step of isolation (cells located in the deeper layers of the valve) ( b ). The composition of stenotic aor tic valve immune infiltrate ( c ) expressed as a percentage (%) of total CD45+ cells, including T helper cells (CD45+, CD4+), T cytotoxic cells (CD45+, CD8+), B cells (CD45+, CD19+), monocytes/macrophages (CD45+, CD11b+, CD14+) and granulocytes (CD45+, CD11b int , CD14−). The rates of ATP hydrolysis, AMP hydrolysis and adenosine deamination on the surface of <t>human</t> monocyte/macrophages (SC; d ) and human <t>peripheral</t> <t>blood</t> <t>mononuclear</t> cells (PBMC = lymphocytes; e ) and in the presence of ecto-enzyme inhibitors. Results are shown as mean ± SEM; n = 5–9, * p
    Human Peripheral Blood Mononuclear Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human peripheral blood mononuclear cells/product/ATCC
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human peripheral blood mononuclear cells - by Bioz Stars, 2021-03
    95/100 stars
      Buy from Supplier

    Image Search Results


    Stenotic aortic valve immune infiltrate is a smaller source of nucleotide-degrading ecto-nucleotidases but a larger of adenosine deaminase. Simplified protocol of stenotic aortic valve cell isolation, including endothelial cells (first step of isolation, vWF positive or CD31 high positive), interstitial cells (second step of isolation; Vimentin positive, Vim+) and immune cells (first and second step of isolation; CD45 positive, CD45+) ( a ). Flow cytometry analysis of CD45 positive cells (immune cells) as a percentage of total isolated cells after first step of isolation (cells located in the upper layers of the valve) and second step of isolation (cells located in the deeper layers of the valve) ( b ). The composition of stenotic aor tic valve immune infiltrate ( c ) expressed as a percentage (%) of total CD45+ cells, including T helper cells (CD45+, CD4+), T cytotoxic cells (CD45+, CD8+), B cells (CD45+, CD19+), monocytes/macrophages (CD45+, CD11b+, CD14+) and granulocytes (CD45+, CD11b int , CD14−). The rates of ATP hydrolysis, AMP hydrolysis and adenosine deamination on the surface of human monocyte/macrophages (SC; d ) and human peripheral blood mononuclear cells (PBMC = lymphocytes; e ) and in the presence of ecto-enzyme inhibitors. Results are shown as mean ± SEM; n = 5–9, * p

    Journal: Clinical Research in Cardiology

    Article Title: Nucleotide ecto-enzyme metabolic pattern and spatial distribution in calcific aortic valve disease; its relation to pathological changes and clinical presentation

    doi: 10.1007/s00392-019-01495-x

    Figure Lengend Snippet: Stenotic aortic valve immune infiltrate is a smaller source of nucleotide-degrading ecto-nucleotidases but a larger of adenosine deaminase. Simplified protocol of stenotic aortic valve cell isolation, including endothelial cells (first step of isolation, vWF positive or CD31 high positive), interstitial cells (second step of isolation; Vimentin positive, Vim+) and immune cells (first and second step of isolation; CD45 positive, CD45+) ( a ). Flow cytometry analysis of CD45 positive cells (immune cells) as a percentage of total isolated cells after first step of isolation (cells located in the upper layers of the valve) and second step of isolation (cells located in the deeper layers of the valve) ( b ). The composition of stenotic aor tic valve immune infiltrate ( c ) expressed as a percentage (%) of total CD45+ cells, including T helper cells (CD45+, CD4+), T cytotoxic cells (CD45+, CD8+), B cells (CD45+, CD19+), monocytes/macrophages (CD45+, CD11b+, CD14+) and granulocytes (CD45+, CD11b int , CD14−). The rates of ATP hydrolysis, AMP hydrolysis and adenosine deamination on the surface of human monocyte/macrophages (SC; d ) and human peripheral blood mononuclear cells (PBMC = lymphocytes; e ) and in the presence of ecto-enzyme inhibitors. Results are shown as mean ± SEM; n = 5–9, * p

    Article Snippet: Isolated human peripheral blood mononuclear cells and monocyte/macrophage cells (SC line, ATCC, cat. CRL-9855) that were used at passage 4, were plated in 24-well cell culture plate at a density 0.2 × 106 per well in a total volume of 1 mL HBSS.

    Techniques: Cell Isolation, Isolation, Flow Cytometry

    Effects of human serum on entry of BCYE- and amoeba-grown L. pneumophila into THP-1 cells (A) and human PBMs (B). Entry is expressed relative to BCYE-grown AA100 in the absence of serum and is calculated as described in Materials and Methods. Serum was replaced by RPMI in the no-serum control. Error bars represent the standard deviations for triplicate samples in each experiment. The results shown are from a single representative experiment. All experiments were repeated at least three times.

    Journal: Infection and Immunity

    Article Title: Intracellular Growth in Acanthamoeba castellanii Affects Monocyte Entry Mechanisms and Enhances Virulence of Legionella pneumophila

    doi:

    Figure Lengend Snippet: Effects of human serum on entry of BCYE- and amoeba-grown L. pneumophila into THP-1 cells (A) and human PBMs (B). Entry is expressed relative to BCYE-grown AA100 in the absence of serum and is calculated as described in Materials and Methods. Serum was replaced by RPMI in the no-serum control. Error bars represent the standard deviations for triplicate samples in each experiment. The results shown are from a single representative experiment. All experiments were repeated at least three times.

    Article Snippet: The monocytes used in these experiments were either human peripheral blood monocytes (PBMs) or THP-1 cells (ATCC TIB202).

    Techniques:

    Cell viability of peripheral blood mononuclear cells (PBMC) after treatment for 24 h. ( A ) Cell viability of PBMC cells after treatment with the ethanolic extract of E. camaldulensis for 24 h. ( B ) Cell viability of PBMC cells after treatment with increasing concentrations of CDDP for 24 h. ( C ) Cell viability of PBMC cells after treatment with the aqueous extract of E. camaldulensis for 24 h. ( D ) Cell viability of PBMC 24 h after treatment with CDDP (4 µg/mL) combined with the aqueous extract (AE 75 µg/mL). ( E ) Cell viability of PBMC 24 h after treatment with CDDP (4 µg/mL) combined with the aqueous extract (AE 150 µg/mL) (* p

    Journal: Medicines

    Article Title: Potency of Combining Eucalyptus camaldulensis subsp. camaldulensis with Low-Dose Cisplatin in A549 Human Lung Adenocarcinomas and MCF-7 Breast Adenocarcinoma

    doi: 10.3390/medicines7080040

    Figure Lengend Snippet: Cell viability of peripheral blood mononuclear cells (PBMC) after treatment for 24 h. ( A ) Cell viability of PBMC cells after treatment with the ethanolic extract of E. camaldulensis for 24 h. ( B ) Cell viability of PBMC cells after treatment with increasing concentrations of CDDP for 24 h. ( C ) Cell viability of PBMC cells after treatment with the aqueous extract of E. camaldulensis for 24 h. ( D ) Cell viability of PBMC 24 h after treatment with CDDP (4 µg/mL) combined with the aqueous extract (AE 75 µg/mL). ( E ) Cell viability of PBMC 24 h after treatment with CDDP (4 µg/mL) combined with the aqueous extract (AE 150 µg/mL) (* p

    Article Snippet: Peripheral Blood Mononuclear Cells (PBMC)Human peripheral blood mononuclear cells (PBMCs) obtained from ATCC, Manassas, VA, USA, comprising lymphocytes (B-cells, T-cells, and NK-cells), monocytes, and dendritic cells, were frequently used for the evaluation of immune responses.

    Techniques:

    S. sonnei induces the secretion of IL-1β, IL-18, NLRP3, ASC, and active caspase-1 in macrophages. (A) J774A.1 macrophages, THP-1 macrophages, PBMCs or BMDM were primed with 1 μg/ml LPS for 4 h and then infected with S. sonnei for an additional 20 h. The levels of IL-1β in the supernatants were measured by ELISA. (B–E) J774A.1 macrophages or THP-1 macrophages were primed with 1 μg/ml LPS for 4 h followed and then infected with S. sonnei for an additional 20 h or stimulated with 5 mM ATP for an additional 0.5 h. The levels of IL-1β (B) , IL-18 (C) , caspase-1 (D) , NLRP3, and ASC (E) in the supernatants were measured by Western blotting. The ELISA data are expressed as the mean ± SD of four separate experiments. The Western blotting results are representative of three different experiments. * and *** indicate significant differences at the levels of p

    Journal: Frontiers in Immunology

    Article Title: Critical Role for the NLRP3 Inflammasome in Mediating IL-1β Production in Shigella sonnei-Infected Macrophages

    doi: 10.3389/fimmu.2020.01115

    Figure Lengend Snippet: S. sonnei induces the secretion of IL-1β, IL-18, NLRP3, ASC, and active caspase-1 in macrophages. (A) J774A.1 macrophages, THP-1 macrophages, PBMCs or BMDM were primed with 1 μg/ml LPS for 4 h and then infected with S. sonnei for an additional 20 h. The levels of IL-1β in the supernatants were measured by ELISA. (B–E) J774A.1 macrophages or THP-1 macrophages were primed with 1 μg/ml LPS for 4 h followed and then infected with S. sonnei for an additional 20 h or stimulated with 5 mM ATP for an additional 0.5 h. The levels of IL-1β (B) , IL-18 (C) , caspase-1 (D) , NLRP3, and ASC (E) in the supernatants were measured by Western blotting. The ELISA data are expressed as the mean ± SD of four separate experiments. The Western blotting results are representative of three different experiments. * and *** indicate significant differences at the levels of p

    Article Snippet: THP-1 macrophages were differentiated from THP-1 monocytes by treatment with 50 nM PMA for 48 h. Human peripheral blood mononuclear cells (PBMCs) were separated from whole blood from healthy volunteers by density gradient centrifugation using Histopaque-1077 , and all experimental protocols were performed in accordance with the guidelines and regulations provided and accepted by the Institutional Review Board of the Tri-Service General Hospital, National Defense Medical Center and the volunteers' informed consent (TSGH-IRB-2-106-05-190 and TSGH-IRB-2-106-05-009).

    Techniques: Infection, Enzyme-linked Immunosorbent Assay, Western Blot