human peripheral blood mononuclear cells  (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 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
    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/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, 2021-04
    95/100 stars

    Images

    1) Product Images from "Nucleotide ecto-enzyme metabolic pattern and spatial distribution in calcific aortic valve disease; its relation to pathological changes and clinical presentation"

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

    Journal: Clinical Research in Cardiology

    doi: 10.1007/s00392-019-01495-x

    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
    Figure Legend 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

    Techniques Used: Cell Isolation, Isolation, Flow Cytometry

    2) Product Images from "Role of H2-calponin in Regulating Macrophage Motility and Phagocytosis *"

    Article Title: Role of H2-calponin in Regulating Macrophage Motility and Phagocytosis *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M801163200

    Expression of h2-calponin in myeloid cells. A , Western blots using anti-h2-calponin polyclonal antibody RAH2 and mAb CP22 on total protein extracted from human peripheral mononuclear cells, human myelogenous leukemia cell line K562, human monocyte line THP-1, and in vitro differentiated human myeloid leukemia cell line HL-60 detected significant amounts of h2-calponin. B , no calponin expression was detectable in the human T lymphocyte line Jurkat, mouse T lymphocyte line BW5147, mouse plasmacytoma cell line T1165, and mouse myeloma cell line NS-1 by RAH2 Western blot.
    Figure Legend Snippet: Expression of h2-calponin in myeloid cells. A , Western blots using anti-h2-calponin polyclonal antibody RAH2 and mAb CP22 on total protein extracted from human peripheral mononuclear cells, human myelogenous leukemia cell line K562, human monocyte line THP-1, and in vitro differentiated human myeloid leukemia cell line HL-60 detected significant amounts of h2-calponin. B , no calponin expression was detectable in the human T lymphocyte line Jurkat, mouse T lymphocyte line BW5147, mouse plasmacytoma cell line T1165, and mouse myeloma cell line NS-1 by RAH2 Western blot.

    Techniques Used: Expressing, Western Blot, In Vitro

    Related Articles

    Incubation:

    Article Title: Salmonella Typhi-specific multifunctional CD8+ T cells play a dominant role in protection from typhoid fever in humans
    Article Snippet: Stimulator cells Autologous Epstein-Barr virus (EBV)-transformed lymphoblastoid cell line (B-EBV cells) and autologous blasts were generated from the PBMC of each participant isolated before challenge. .. B-EBV cells were obtained by incubation of the PBMC with EBV-containing supernatant from the B95-8 cell line (ATCC CRL1612) and cyclosporine (0.5 ug/mL; Sigma-Aldrich, Saint-Louis, MO) at 37 °C with 5 % CO2 for 2–3 weeks. .. PHA-activated blasts were prepared by incubating PBMC with 1 μg/ml PHA (Sigma-Aldrich, St. Louis, MO) in cRPMI for 24 h, followed by washing and culture in cRPMI containing 20 IU/ml recombinant human IL-2 (rhIL-2; Roche, Indianapolis, IN) for 7 days.

    Article Title: A “possible” involvement of TNF-alpha in apoptosis induction in peripheral blood lymphocytes of cats with feline infectious peritonitis
    Article Snippet: .. 2.6 Separation of CD8+ , CD4+ , and CD21+ cells To separate CD8+ cells, PBMC (1 × 107 cells) were incubated with 500 μl of purified monoclonal antibody (MAb) OKT8 (ATCC CRL8014) IgG at 4 °C for 30 min. MAb OKT8 recognizes the alpha-chain of human CD8 antigen and cross-reacts with feline CD8+ cells ( , ). .. The cells were washed three times with PBS containing 2 mM EDTA and 0.5% BSA, 40 μl of microbeads coated with rat anti-mouse IgG2a+b (Miltenyi Biotec, Germany) were added to the cells, and the mixture was incubated at 4 °C for 15 min. After washing with PBS containing 2 mM EDTA and 0.5% BSA, the cells were fractionated into CD8+ cells and CD8+ -depleted cells by a magnetic system using a MACS kit (Miltenyi Biotec, Germany).

    Isolation:

    Article Title: Activation of TAFI on the Surface of Streptococcus pyogenes Evokes Inflammatory Reactions by Modulating the Kallikrein/Kinin System
    Article Snippet: .. Human peripheral blood mononuclear cells (PBMCs) were isolated as previously described [ ]. .. Stimulation of PBMCs PBMCs were incubated with 1% (v/v) S. pyogenes (M41) supernatants (obtained from overnight cultures of single colonies in 40 ml TH medium) in RPMI 1640 medium (Invitrogen, Paisley, UK) in the presence of 2 mM l-glutamine for 24 h at 37°C.

    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.

    Purification:

    Article Title: A “possible” involvement of TNF-alpha in apoptosis induction in peripheral blood lymphocytes of cats with feline infectious peritonitis
    Article Snippet: .. 2.6 Separation of CD8+ , CD4+ , and CD21+ cells To separate CD8+ cells, PBMC (1 × 107 cells) were incubated with 500 μl of purified monoclonal antibody (MAb) OKT8 (ATCC CRL8014) IgG at 4 °C for 30 min. MAb OKT8 recognizes the alpha-chain of human CD8 antigen and cross-reacts with feline CD8+ cells ( , ). .. The cells were washed three times with PBS containing 2 mM EDTA and 0.5% BSA, 40 μl of microbeads coated with rat anti-mouse IgG2a+b (Miltenyi Biotec, Germany) were added to the cells, and the mixture was incubated at 4 °C for 15 min. After washing with PBS containing 2 mM EDTA and 0.5% BSA, the cells were fractionated into CD8+ cells and CD8+ -depleted cells by a magnetic system using a MACS kit (Miltenyi Biotec, Germany).

    Multiple Displacement Amplification:

    Article Title: An Fc engineering approach that modulates antibody-dependent cytokine release without altering cell-killing functions
    Article Snippet: Data were log transformed and fitted to a sigmoidal dose-response curve using GraphPad Prism v5. .. PBMC-based ADCR ADCR assays with PBMC effector cells and MDA-MB-231 target cells (from American Type Culture Collection) were performed using an effector: target ratio of 10:1 (200,000 PBMC: 20,000 target cells) in a 96-well plate in the presence or absence of 10 ng/ml IL-12 (R & D Systems, cat.no. .. 219-IL-005) with the indicated antibody concentrations.

    Chloramphenicol Acetyltransferase Assay:

    Article Title: An Fc engineering approach that modulates antibody-dependent cytokine release without altering cell-killing functions
    Article Snippet: Data were log transformed and fitted to a sigmoidal dose-response curve using GraphPad Prism v5. .. PBMC-based ADCR ADCR assays with PBMC effector cells and MDA-MB-231 target cells (from American Type Culture Collection) were performed using an effector: target ratio of 10:1 (200,000 PBMC: 20,000 target cells) in a 96-well plate in the presence or absence of 10 ng/ml IL-12 (R & D Systems, cat.no. .. 219-IL-005) with the indicated antibody concentrations.

    Cell Culture:

    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.

    Microarray:

    Article Title: Enterovirus-associated changes in blood transcriptomic profiles of children with genetic susceptibility to type 1 diabetes
    Article Snippet: .. Microarray data from human PBMCs infected in vitro The transcriptomics data from children at risk for type 1 diabetes were compared with microarray data (HumanHT-12 V3.0 BeadChip, Illumina, San Diego, CA, USA) from peripheral blood mononuclear cells (PBMCs) infected with enterovirus in vitro [ ], including three replicate samples of PBMCs infected with ATCC strain of echovirus 9 or wild-type Coxsackie B1 virus strains CDC10802 and CDC10796 for 48 h, and uninfected control PBMCs (see Dataset 1 published on https://www.btk.fi/1234-2/ ). ..

    Infection:

    Article Title: Enterovirus-associated changes in blood transcriptomic profiles of children with genetic susceptibility to type 1 diabetes
    Article Snippet: .. Microarray data from human PBMCs infected in vitro The transcriptomics data from children at risk for type 1 diabetes were compared with microarray data (HumanHT-12 V3.0 BeadChip, Illumina, San Diego, CA, USA) from peripheral blood mononuclear cells (PBMCs) infected with enterovirus in vitro [ ], including three replicate samples of PBMCs infected with ATCC strain of echovirus 9 or wild-type Coxsackie B1 virus strains CDC10802 and CDC10796 for 48 h, and uninfected control PBMCs (see Dataset 1 published on https://www.btk.fi/1234-2/ ). ..

    In Vitro:

    Article Title: Enterovirus-associated changes in blood transcriptomic profiles of children with genetic susceptibility to type 1 diabetes
    Article Snippet: .. Microarray data from human PBMCs infected in vitro The transcriptomics data from children at risk for type 1 diabetes were compared with microarray data (HumanHT-12 V3.0 BeadChip, Illumina, San Diego, CA, USA) from peripheral blood mononuclear cells (PBMCs) infected with enterovirus in vitro [ ], including three replicate samples of PBMCs infected with ATCC strain of echovirus 9 or wild-type Coxsackie B1 virus strains CDC10802 and CDC10796 for 48 h, and uninfected control PBMCs (see Dataset 1 published on https://www.btk.fi/1234-2/ ). ..

    Activation Assay:

    Article Title: Protein Kinase D1: A New Component in Toll-Like Receptor 9 Signaling
    Article Snippet: .. In addition, CpG-B DNA and EC DNA induced activation of PKD family proteins in human PBMCs. ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    ATCC blood mononuclear cells pbmcs
    Expression of the 77 genes present in our enterovirus-induced signature and upregulated in all three in vitro enterovirus infections in <t>microarray</t> data published by: ( a ) Kallionpää et al [ 15 ]; and ( b ) Ferreira et al [ 27 ]. Sums of child-specific z scores over the 77 genes were calculated for each of the 356 whole blood samples by Kallionpää et al [ 15 ] (GEO: GSE30211) and the 454 <t>PBMC</t> samples by Ferreira et al [ 27 ] (Array Express: E-MTAB-1724), as described in Methods, using the published pre-processed datasets and sample information based on personal communications with Ferreira et al. All probes ( a ) or the highest-intensity exons mapping to genes ( b ) overlapping with the 77 genes were summed. ( a ) Black, strongly enterovirus-positive blood samples; white, weakly enterovirus-positive blood samples; grey, enterovirus-negative blood samples. ( a , b ) PreSero, samples collected from before seroconversion from children with autoantibody positivity or type 1 diabetes (in a , n = 22; in b , n = 65); PostSero, samples collected after seroconversion from children with autoantibody positivity or type 1 diabetes (in a , n = 169; b n = 84), Aab − , samples collected from autoantibody-negative children (in a , n = 165; in b , n = 305). ( c ) Venn diagram showing the overlaps between the 77 genes present in the enterovirus-induced signature and upregulated in all three in vitro enterovirus infections; genes upregulated before or after seroconversion in autoantibody-positive children based on the results by Kallionpää et al [ 15 ] and the 225 interferon-inducible genes detected by Ferreira et al [ 27 ]
    Blood Mononuclear Cells Pbmcs, 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/blood mononuclear cells pbmcs/product/ATCC
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    blood mononuclear cells pbmcs - by Bioz Stars, 2021-04
    95/100 stars
      Buy from Supplier

    Image Search Results


    Expression of the 77 genes present in our enterovirus-induced signature and upregulated in all three in vitro enterovirus infections in microarray data published by: ( a ) Kallionpää et al [ 15 ]; and ( b ) Ferreira et al [ 27 ]. Sums of child-specific z scores over the 77 genes were calculated for each of the 356 whole blood samples by Kallionpää et al [ 15 ] (GEO: GSE30211) and the 454 PBMC samples by Ferreira et al [ 27 ] (Array Express: E-MTAB-1724), as described in Methods, using the published pre-processed datasets and sample information based on personal communications with Ferreira et al. All probes ( a ) or the highest-intensity exons mapping to genes ( b ) overlapping with the 77 genes were summed. ( a ) Black, strongly enterovirus-positive blood samples; white, weakly enterovirus-positive blood samples; grey, enterovirus-negative blood samples. ( a , b ) PreSero, samples collected from before seroconversion from children with autoantibody positivity or type 1 diabetes (in a , n = 22; in b , n = 65); PostSero, samples collected after seroconversion from children with autoantibody positivity or type 1 diabetes (in a , n = 169; b n = 84), Aab − , samples collected from autoantibody-negative children (in a , n = 165; in b , n = 305). ( c ) Venn diagram showing the overlaps between the 77 genes present in the enterovirus-induced signature and upregulated in all three in vitro enterovirus infections; genes upregulated before or after seroconversion in autoantibody-positive children based on the results by Kallionpää et al [ 15 ] and the 225 interferon-inducible genes detected by Ferreira et al [ 27 ]

    Journal: Diabetologia

    Article Title: Enterovirus-associated changes in blood transcriptomic profiles of children with genetic susceptibility to type 1 diabetes

    doi: 10.1007/s00125-017-4460-7

    Figure Lengend Snippet: Expression of the 77 genes present in our enterovirus-induced signature and upregulated in all three in vitro enterovirus infections in microarray data published by: ( a ) Kallionpää et al [ 15 ]; and ( b ) Ferreira et al [ 27 ]. Sums of child-specific z scores over the 77 genes were calculated for each of the 356 whole blood samples by Kallionpää et al [ 15 ] (GEO: GSE30211) and the 454 PBMC samples by Ferreira et al [ 27 ] (Array Express: E-MTAB-1724), as described in Methods, using the published pre-processed datasets and sample information based on personal communications with Ferreira et al. All probes ( a ) or the highest-intensity exons mapping to genes ( b ) overlapping with the 77 genes were summed. ( a ) Black, strongly enterovirus-positive blood samples; white, weakly enterovirus-positive blood samples; grey, enterovirus-negative blood samples. ( a , b ) PreSero, samples collected from before seroconversion from children with autoantibody positivity or type 1 diabetes (in a , n = 22; in b , n = 65); PostSero, samples collected after seroconversion from children with autoantibody positivity or type 1 diabetes (in a , n = 169; b n = 84), Aab − , samples collected from autoantibody-negative children (in a , n = 165; in b , n = 305). ( c ) Venn diagram showing the overlaps between the 77 genes present in the enterovirus-induced signature and upregulated in all three in vitro enterovirus infections; genes upregulated before or after seroconversion in autoantibody-positive children based on the results by Kallionpää et al [ 15 ] and the 225 interferon-inducible genes detected by Ferreira et al [ 27 ]

    Article Snippet: Microarray data from human PBMCs infected in vitro The transcriptomics data from children at risk for type 1 diabetes were compared with microarray data (HumanHT-12 V3.0 BeadChip, Illumina, San Diego, CA, USA) from peripheral blood mononuclear cells (PBMCs) infected with enterovirus in vitro [ ], including three replicate samples of PBMCs infected with ATCC strain of echovirus 9 or wild-type Coxsackie B1 virus strains CDC10802 and CDC10796 for 48 h, and uninfected control PBMCs (see Dataset 1 published on https://www.btk.fi/1234-2/ ).

    Techniques: Expressing, In Vitro, Microarray

    IgG1 antibodies mediate cytotoxicity and IFNγ cytokine secretion by PBMCs. ( A ) ADCC was performed using human PBMC as effector cells and MDA-MB-231 cells as targets with the indicated concentrations of anti-CD142 antibodies. Data are representative of 2 independent experiments (duplicate measurements per experiment). ( B–C ) ADCR assays were performed using PBMC effector cells and MDA-MB-231 target cells in the absence ( B ) or the presence ( C ) of IL-12. After 48-h supernatants were collected and IFNγ secretion was measured using ELISA. Data are representative of 2 independent experiments (duplicate measurements per experiment).

    Journal: mAbs

    Article Title: An Fc engineering approach that modulates antibody-dependent cytokine release without altering cell-killing functions

    doi: 10.1080/19420862.2015.1022692

    Figure Lengend Snippet: IgG1 antibodies mediate cytotoxicity and IFNγ cytokine secretion by PBMCs. ( A ) ADCC was performed using human PBMC as effector cells and MDA-MB-231 cells as targets with the indicated concentrations of anti-CD142 antibodies. Data are representative of 2 independent experiments (duplicate measurements per experiment). ( B–C ) ADCR assays were performed using PBMC effector cells and MDA-MB-231 target cells in the absence ( B ) or the presence ( C ) of IL-12. After 48-h supernatants were collected and IFNγ secretion was measured using ELISA. Data are representative of 2 independent experiments (duplicate measurements per experiment).

    Article Snippet: PBMC-based ADCR ADCR assays with PBMC effector cells and MDA-MB-231 target cells (from American Type Culture Collection) were performed using an effector: target ratio of 10:1 (200,000 PBMC: 20,000 target cells) in a 96-well plate in the presence or absence of 10 ng/ml IL-12 (R & D Systems, cat.no.

    Techniques: Multiple Displacement Amplification, Enzyme-linked Immunosorbent Assay

    Macrophage-mediated ADCR is dependent on interactions with FcγRI. ( A ) Macrophage 24-h tumor cell-killing is depicted for 4 independent donors (duplicate measurements per experiment). ( B ) At the end of the 24-h incubation in the tumor cell-killing assay, supernatants were collected and analyzed for cytokine levels. Dot plots on top represent the concentration of cytokines detected from a single donor (duplicate measurements per experiment), and the bar graphs below represent the mean +/− SEM fold-change in cytokine levels normalized to the isotype control from 4 independent PBMC donors.

    Journal: mAbs

    Article Title: An Fc engineering approach that modulates antibody-dependent cytokine release without altering cell-killing functions

    doi: 10.1080/19420862.2015.1022692

    Figure Lengend Snippet: Macrophage-mediated ADCR is dependent on interactions with FcγRI. ( A ) Macrophage 24-h tumor cell-killing is depicted for 4 independent donors (duplicate measurements per experiment). ( B ) At the end of the 24-h incubation in the tumor cell-killing assay, supernatants were collected and analyzed for cytokine levels. Dot plots on top represent the concentration of cytokines detected from a single donor (duplicate measurements per experiment), and the bar graphs below represent the mean +/− SEM fold-change in cytokine levels normalized to the isotype control from 4 independent PBMC donors.

    Article Snippet: PBMC-based ADCR ADCR assays with PBMC effector cells and MDA-MB-231 target cells (from American Type Culture Collection) were performed using an effector: target ratio of 10:1 (200,000 PBMC: 20,000 target cells) in a 96-well plate in the presence or absence of 10 ng/ml IL-12 (R & D Systems, cat.no.

    Techniques: Incubation, Concentration Assay

    ( See previous page ). Multiple cytokines are triggered by engagement of FcγRs on PBMCs in response to antibody-opsonized tumor cells. ADCR assays with PBMC effector cells were performed in the absence ( A ) or presence ( B ) of IL-12. After 48 h, supernatants were collected and analyzed by a quantitative Luminex Multi-Analyte Profile assay. Dot plots on top represent the concentration of cytokines detected from a single donor, and the bar graphs below represent the mean +/− SEM fold-change in cytokine levels normalized to the isotype control from 4 independent PBMC donors. ( C ) Shown are flow cytometric histograms of IFNγ secretion elicited by IgG1 (black) compared to negative control (gray) of NK Cells (CD56 pos , CD3 neg ), NKT cells (CD56 pos CD3 pos ), CD16 pos monocytes (CD14 pos CD16 pos ), CD16 neg monocytes (CD14 pos CD16 neg ), T cells (CD3 pos ). ( D ) NK and NKT cells were the primary producers of IFNγ in ADCR assays using PBMCs. ADCR assays were performed with PBMCs, MDA-MB-231-GFP target cells and 5 µg/ml of anti-CD142 antibody variants in the absence or presence of IL-12. Bar graphs summarize the percent of each population secreting IFNγ elicited by each of the antibody variants. Data are representative from 3 donors in 2 independent experiments.

    Journal: mAbs

    Article Title: An Fc engineering approach that modulates antibody-dependent cytokine release without altering cell-killing functions

    doi: 10.1080/19420862.2015.1022692

    Figure Lengend Snippet: ( See previous page ). Multiple cytokines are triggered by engagement of FcγRs on PBMCs in response to antibody-opsonized tumor cells. ADCR assays with PBMC effector cells were performed in the absence ( A ) or presence ( B ) of IL-12. After 48 h, supernatants were collected and analyzed by a quantitative Luminex Multi-Analyte Profile assay. Dot plots on top represent the concentration of cytokines detected from a single donor, and the bar graphs below represent the mean +/− SEM fold-change in cytokine levels normalized to the isotype control from 4 independent PBMC donors. ( C ) Shown are flow cytometric histograms of IFNγ secretion elicited by IgG1 (black) compared to negative control (gray) of NK Cells (CD56 pos , CD3 neg ), NKT cells (CD56 pos CD3 pos ), CD16 pos monocytes (CD14 pos CD16 pos ), CD16 neg monocytes (CD14 pos CD16 neg ), T cells (CD3 pos ). ( D ) NK and NKT cells were the primary producers of IFNγ in ADCR assays using PBMCs. ADCR assays were performed with PBMCs, MDA-MB-231-GFP target cells and 5 µg/ml of anti-CD142 antibody variants in the absence or presence of IL-12. Bar graphs summarize the percent of each population secreting IFNγ elicited by each of the antibody variants. Data are representative from 3 donors in 2 independent experiments.

    Article Snippet: PBMC-based ADCR ADCR assays with PBMC effector cells and MDA-MB-231 target cells (from American Type Culture Collection) were performed using an effector: target ratio of 10:1 (200,000 PBMC: 20,000 target cells) in a 96-well plate in the presence or absence of 10 ng/ml IL-12 (R & D Systems, cat.no.

    Techniques: Polyacrylamide Gel Electrophoresis, Luminex, Concentration Assay, Flow Cytometry, Negative Control, Multiple Displacement Amplification

    CpG-B DNA, not CpG-A DNA, induces phosphorylation of PKD proteins in murine splenic B cells, cDCs, and pDCs, and human PBMCs

    Journal:

    Article Title: Protein Kinase D1: A New Component in Toll-Like Receptor 9 Signaling

    doi:

    Figure Lengend Snippet: CpG-B DNA, not CpG-A DNA, induces phosphorylation of PKD proteins in murine splenic B cells, cDCs, and pDCs, and human PBMCs

    Article Snippet: In addition, CpG-B DNA and EC DNA induced activation of PKD family proteins in human PBMCs.

    Techniques:

    In vitro effects of the culture supernatant of macrophages from FIP cats on the expression of TNFR1 and TNFR2 mRNA in CD4 + , CD8 + , and CD21 + cells. CD4 + , CD8 + , and CD21 + cells were recovered from the PBMC of SPF cats using magnetic beads, and cultured in the presence of the culture supernatant of PEC from FIP cats. The levels of TNFR1 and TNFR2 mRNA expression were measured 0 and 2 h after culture. The levels of TNFR1 and TNFR2 mRNA expression were quantitatively analyzed in terms of the relative density value to the mRNA for the housekeeping gene GAPDH. ** p

    Journal: Veterinary Microbiology

    Article Title: A “possible” involvement of TNF-alpha in apoptosis induction in peripheral blood lymphocytes of cats with feline infectious peritonitis

    doi: 10.1016/j.vetmic.2006.08.033

    Figure Lengend Snippet: In vitro effects of the culture supernatant of macrophages from FIP cats on the expression of TNFR1 and TNFR2 mRNA in CD4 + , CD8 + , and CD21 + cells. CD4 + , CD8 + , and CD21 + cells were recovered from the PBMC of SPF cats using magnetic beads, and cultured in the presence of the culture supernatant of PEC from FIP cats. The levels of TNFR1 and TNFR2 mRNA expression were measured 0 and 2 h after culture. The levels of TNFR1 and TNFR2 mRNA expression were quantitatively analyzed in terms of the relative density value to the mRNA for the housekeeping gene GAPDH. ** p

    Article Snippet: 2.6 Separation of CD8+ , CD4+ , and CD21+ cells To separate CD8+ cells, PBMC (1 × 107 cells) were incubated with 500 μl of purified monoclonal antibody (MAb) OKT8 (ATCC CRL8014) IgG at 4 °C for 30 min. MAb OKT8 recognizes the alpha-chain of human CD8 antigen and cross-reacts with feline CD8+ cells ( , ).

    Techniques: In Vitro, Expressing, Magnetic Beads, Cell Culture

    Apoptosis-inducing activities of specimens from FIP cats in peripheral blood CD4 + , CD8 + , and CD21 + cells. The PBMC (2 × 10 6 ) of SPF cats and their subsets of lymphocytes CD4 + , CD8 + , and CD21 + cells (2 × 10 6 ) were cultured at 37 °C for 4 h in the presence of the ascitic fluid, PEC culture supernatant, or plasma of FIP cats, and apoptotic cells were detected by TUNEL. CD4 + , CD8 + , and CD21 + cells were recovered with magnetic beads. Medium (white), ascitic fluid of FIP cats (gray), culture supernatant of PEC from FIP cats (oblique), plasma of FIP cats (dot), plasma of FIP-infected non-FIP cats (black).

    Journal: Veterinary Microbiology

    Article Title: A “possible” involvement of TNF-alpha in apoptosis induction in peripheral blood lymphocytes of cats with feline infectious peritonitis

    doi: 10.1016/j.vetmic.2006.08.033

    Figure Lengend Snippet: Apoptosis-inducing activities of specimens from FIP cats in peripheral blood CD4 + , CD8 + , and CD21 + cells. The PBMC (2 × 10 6 ) of SPF cats and their subsets of lymphocytes CD4 + , CD8 + , and CD21 + cells (2 × 10 6 ) were cultured at 37 °C for 4 h in the presence of the ascitic fluid, PEC culture supernatant, or plasma of FIP cats, and apoptotic cells were detected by TUNEL. CD4 + , CD8 + , and CD21 + cells were recovered with magnetic beads. Medium (white), ascitic fluid of FIP cats (gray), culture supernatant of PEC from FIP cats (oblique), plasma of FIP cats (dot), plasma of FIP-infected non-FIP cats (black).

    Article Snippet: 2.6 Separation of CD8+ , CD4+ , and CD21+ cells To separate CD8+ cells, PBMC (1 × 107 cells) were incubated with 500 μl of purified monoclonal antibody (MAb) OKT8 (ATCC CRL8014) IgG at 4 °C for 30 min. MAb OKT8 recognizes the alpha-chain of human CD8 antigen and cross-reacts with feline CD8+ cells ( , ).

    Techniques: Cell Culture, TUNEL Assay, Magnetic Beads, Infection