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BioLegend anti mouse igg fitc pe
Anti Mouse Igg Fitc Pe, supplied by BioLegend, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti mouse igg fitc pe - by Bioz Stars, 2021-07
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Staining:

Article Title: OSU-T315 as an Interesting Lead Molecule for Novel B Cell-Specific Therapeutics
Article Snippet: Distinction between the homozygous and the heterozygous CD19-Cre mice was done by flow cytometry on murine peripheral blood ( ). .. Murine blood cells were stained with FITC-labelled anti-murine CD19 (Biolegend, ImTec Diagnostics N.V., Antwerp, Belgium) and PE/Cy5-labelled anti-murine CD45R/B220 (Biolegend, ImTec Diagnostics N.V., Antwerp, Belgium) and analyzed with the 3-color Becton Dickinson FACSCalibur apparatus. .. CD45R/B220, commonly used as a pan-B cell marker, was included in the double staining to allow visualization of B cells in the blood of mice with lowered (heterozygous CD19-Cre genotype) or completely deleted (homozygous CD19-Cre genotype) expression of CD19.

Article Title: Leptospira surface adhesin (Lsa21) induces Toll like receptor 2 and 4 mediated inflammatory responses in macrophages
Article Snippet: .. Then the cells were stained by sequential 1-h incubations with 1:100 dilution of mouse anti-Lsa21 (mouse serum) and FITC or Alexa Flour 647 conjugated rabbit anti-mouse IgG (Biolegend, USA), PE-conjugated rabbit anti-mouse TLR2 and APC conjugated rabbit anti-mouse TLR4. .. Between each staining step, the cells were washed three times with 1% BSA in PBS.

Article Title: Toll-like receptor 4 inhibition prevents autoimmune diabetes in NOD mice
Article Snippet: For antigen-specific activation assay, BDC2.5 splenocytes were stained with 2.5μM CFSE (Life Technologies, Carlsbad, USA) following the manufacturer’s instructions and were stimulated with 0.05, 0.2, 0.5 and 1 μg/ml of BDC2.5 mimotope, in presence or absence of 10μg/ml CLI-095 for 48 h. Proliferation was analysed by CFSE dilution. .. Flow cytometry Lymphocytes were stained with the following fluorochrome-conjugated antibodies as indicated in the manufacturer’s instructions: FITC anti-mouse CD25 antibody (3C7, Biolegend, San Diego, USA), PE/Cy7 anti-mouse CD69 antibody (H1.2F3, Biolegend, San Diego, USA), PE anti-mouse CD69 antibody (H1.2F3, Biolegend, San Diego, USA), PE/Cy7 anti-mouse CD8a antibody (53-6.7, Biolegend, San Diego, USA), PE anti-mouse CD8a antibody (53-6.7, Biolegend, San Diego, USA), APC/Cy7 anti-mouse CD8a antibody (53-6.7, Biolegend, San Diego, USA), APC anti-mouse CD8a antibody (53-6.7, Biolegend, San Diego, USA), PerCP anti-mouse CD4 antibody (RM4-5, Biolegend, San Diego, USA), PE/Dazzle™ 594 anti-mouse CD4 antibody (GK1.5, Biolegend, San Diego, USA), PE anti-mouse CD3 antibody (17A2, Biolegend, San Diego, USA), PE anti-mouse TCR β chain antibody (H57-597, Biolegend, San Diego, USA), PE anti-mouse CD62L antibody (MEL-14, Biolegend, San Diego, USA), were used. .. The following isotype control antibodies were used: FITC Rat IgG1 κ isotype Ctrl antibody (RTK2071, Biolegend, San Diego, USA), PE Rat IgG2b κ isotype Ctrl antibody (RTK4530, Biolegend, San Diego, USA), FITC Rat IgG2b κ isotype Ctrl antibody (RTK4530, Biolegend, San Diego, USA), PE Rat IgG2a κ isotype Ctrl antibody (RTK2758, Biolegend, San Diego, USA), PE Armenian Hamster IgG isotype Ctrl antibody (HTK888, Biolegend, San Diego, USA), PE/Cy7 Rat IgG1 κ isotype Ctrl antibody (RTK2071, Biolegend, San Diego, USA).

Flow Cytometry:

Article Title: Antibody-secreting plasma cells persist for decades in human intestine
Article Snippet: .. Flow cytometry and FACS Single-cell suspensions from lamina propria were analyzed by flow cytometry according to standard procedure using fluorochrome-conjugated antibodies targeting HLA-A2–PE (Abcam); CD20-FITC, CD45-APC-H7, and Ki67-FITC (BD); Bcl2–Alexa Fluor 488, BCMA-PE, CD3-APC, CD14-APC, CD19–Alexa Flour 488/PE-Cy7, CD20-PE, CD27-BV421/Pacific blue/APC-Cy7/PE-Cy7, CD28–Alexa Fluor 488, CD38-PE/APC-Cy7, CD45-BV510/APC-Cy7, CD56–Alexa Flour 488, CD95-PE, and HLA-DR–BV605/PerCP (BioLegend); CD138-FITC/PE-Cy7 and HLA-A3–FITC/APC (eBioscience); HLA-B7–PE (EMD Millipore); HLA-B8–PE (Miltenyi Biotec); IgA-FITC/PE, IgM-FITC/PE, and IgD-FITC/PE (SouthernBiotech); and Blimp-1–DyLight 488 (Thermo Fisher Scientific). .. For detection of intracellular antigens, cells were fixed in formaldehyde and permeabilized using a Foxp3 staining buffer set (eBioscience).

Article Title: Lipocalin-2 Functions as Inhibitor of Innate Resistance to Mycobacterium tuberculosis
Article Snippet: .. Total B-cell, T-cell, and neutrophil cell counts were estimated by flow cytometry on total lung homogenate 3 and 5 weeks post-challenge using anti-Ly6G APC (clone 1A8, Biolegend), anti-CD11b PE (clone M1/79, Biolegend), anti-CD11c PerCP-Cy5.5 (clone HL3, BD), anti-CD19 APC (clone 1D3, BD), anti-CD4 FITC (clone RM4-5, BD), and anti-CD8 PE (clone 53-6.7, BD). .. Flow cytometry was performed on BD Accuri C6 and analyzed with FlowJo 10.0.8 software (Tree Star Inc).

Article Title: Toll-like receptor 4 inhibition prevents autoimmune diabetes in NOD mice
Article Snippet: For antigen-specific activation assay, BDC2.5 splenocytes were stained with 2.5μM CFSE (Life Technologies, Carlsbad, USA) following the manufacturer’s instructions and were stimulated with 0.05, 0.2, 0.5 and 1 μg/ml of BDC2.5 mimotope, in presence or absence of 10μg/ml CLI-095 for 48 h. Proliferation was analysed by CFSE dilution. .. Flow cytometry Lymphocytes were stained with the following fluorochrome-conjugated antibodies as indicated in the manufacturer’s instructions: FITC anti-mouse CD25 antibody (3C7, Biolegend, San Diego, USA), PE/Cy7 anti-mouse CD69 antibody (H1.2F3, Biolegend, San Diego, USA), PE anti-mouse CD69 antibody (H1.2F3, Biolegend, San Diego, USA), PE/Cy7 anti-mouse CD8a antibody (53-6.7, Biolegend, San Diego, USA), PE anti-mouse CD8a antibody (53-6.7, Biolegend, San Diego, USA), APC/Cy7 anti-mouse CD8a antibody (53-6.7, Biolegend, San Diego, USA), APC anti-mouse CD8a antibody (53-6.7, Biolegend, San Diego, USA), PerCP anti-mouse CD4 antibody (RM4-5, Biolegend, San Diego, USA), PE/Dazzle™ 594 anti-mouse CD4 antibody (GK1.5, Biolegend, San Diego, USA), PE anti-mouse CD3 antibody (17A2, Biolegend, San Diego, USA), PE anti-mouse TCR β chain antibody (H57-597, Biolegend, San Diego, USA), PE anti-mouse CD62L antibody (MEL-14, Biolegend, San Diego, USA), were used. .. The following isotype control antibodies were used: FITC Rat IgG1 κ isotype Ctrl antibody (RTK2071, Biolegend, San Diego, USA), PE Rat IgG2b κ isotype Ctrl antibody (RTK4530, Biolegend, San Diego, USA), FITC Rat IgG2b κ isotype Ctrl antibody (RTK4530, Biolegend, San Diego, USA), PE Rat IgG2a κ isotype Ctrl antibody (RTK2758, Biolegend, San Diego, USA), PE Armenian Hamster IgG isotype Ctrl antibody (HTK888, Biolegend, San Diego, USA), PE/Cy7 Rat IgG1 κ isotype Ctrl antibody (RTK2071, Biolegend, San Diego, USA).

Article Title: Interaction between CD177 and platelet endothelial cell adhesion molecule-1 downregulates membrane-bound proteinase-3 (PR3) expression on neutrophils and attenuates neutrophil activation induced by PR3-ANCA
Article Snippet: For indirect enzyme-linked immunosorbent assay (ELISA), mouse anti-human CD177 antibody was purchased from Sino Biological Inc. and horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG was purchased from Abcam (Cambridge, UK). .. For flow cytometry analysis, phycoerythrin (PE)-conjugated mouse monoclonal antibody against human CD177 and the isotype control mouse IgG1 were purchased from BioLegend (San Diego, CA, USA). .. Fluorescein isothiocyanate (FITC)-conjugated mouse monoclonal antibody against human PR3 and the isotype control mouse IgG1 were purchased from Abcam.

Cytometry:

Article Title: Antibody-secreting plasma cells persist for decades in human intestine
Article Snippet: .. Flow cytometry and FACS Single-cell suspensions from lamina propria were analyzed by flow cytometry according to standard procedure using fluorochrome-conjugated antibodies targeting HLA-A2–PE (Abcam); CD20-FITC, CD45-APC-H7, and Ki67-FITC (BD); Bcl2–Alexa Fluor 488, BCMA-PE, CD3-APC, CD14-APC, CD19–Alexa Flour 488/PE-Cy7, CD20-PE, CD27-BV421/Pacific blue/APC-Cy7/PE-Cy7, CD28–Alexa Fluor 488, CD38-PE/APC-Cy7, CD45-BV510/APC-Cy7, CD56–Alexa Flour 488, CD95-PE, and HLA-DR–BV605/PerCP (BioLegend); CD138-FITC/PE-Cy7 and HLA-A3–FITC/APC (eBioscience); HLA-B7–PE (EMD Millipore); HLA-B8–PE (Miltenyi Biotec); IgA-FITC/PE, IgM-FITC/PE, and IgD-FITC/PE (SouthernBiotech); and Blimp-1–DyLight 488 (Thermo Fisher Scientific). .. For detection of intracellular antigens, cells were fixed in formaldehyde and permeabilized using a Foxp3 staining buffer set (eBioscience).

Article Title: Lipocalin-2 Functions as Inhibitor of Innate Resistance to Mycobacterium tuberculosis
Article Snippet: .. Total B-cell, T-cell, and neutrophil cell counts were estimated by flow cytometry on total lung homogenate 3 and 5 weeks post-challenge using anti-Ly6G APC (clone 1A8, Biolegend), anti-CD11b PE (clone M1/79, Biolegend), anti-CD11c PerCP-Cy5.5 (clone HL3, BD), anti-CD19 APC (clone 1D3, BD), anti-CD4 FITC (clone RM4-5, BD), and anti-CD8 PE (clone 53-6.7, BD). .. Flow cytometry was performed on BD Accuri C6 and analyzed with FlowJo 10.0.8 software (Tree Star Inc).

Article Title: Toll-like receptor 4 inhibition prevents autoimmune diabetes in NOD mice
Article Snippet: For antigen-specific activation assay, BDC2.5 splenocytes were stained with 2.5μM CFSE (Life Technologies, Carlsbad, USA) following the manufacturer’s instructions and were stimulated with 0.05, 0.2, 0.5 and 1 μg/ml of BDC2.5 mimotope, in presence or absence of 10μg/ml CLI-095 for 48 h. Proliferation was analysed by CFSE dilution. .. Flow cytometry Lymphocytes were stained with the following fluorochrome-conjugated antibodies as indicated in the manufacturer’s instructions: FITC anti-mouse CD25 antibody (3C7, Biolegend, San Diego, USA), PE/Cy7 anti-mouse CD69 antibody (H1.2F3, Biolegend, San Diego, USA), PE anti-mouse CD69 antibody (H1.2F3, Biolegend, San Diego, USA), PE/Cy7 anti-mouse CD8a antibody (53-6.7, Biolegend, San Diego, USA), PE anti-mouse CD8a antibody (53-6.7, Biolegend, San Diego, USA), APC/Cy7 anti-mouse CD8a antibody (53-6.7, Biolegend, San Diego, USA), APC anti-mouse CD8a antibody (53-6.7, Biolegend, San Diego, USA), PerCP anti-mouse CD4 antibody (RM4-5, Biolegend, San Diego, USA), PE/Dazzle™ 594 anti-mouse CD4 antibody (GK1.5, Biolegend, San Diego, USA), PE anti-mouse CD3 antibody (17A2, Biolegend, San Diego, USA), PE anti-mouse TCR β chain antibody (H57-597, Biolegend, San Diego, USA), PE anti-mouse CD62L antibody (MEL-14, Biolegend, San Diego, USA), were used. .. The following isotype control antibodies were used: FITC Rat IgG1 κ isotype Ctrl antibody (RTK2071, Biolegend, San Diego, USA), PE Rat IgG2b κ isotype Ctrl antibody (RTK4530, Biolegend, San Diego, USA), FITC Rat IgG2b κ isotype Ctrl antibody (RTK4530, Biolegend, San Diego, USA), PE Rat IgG2a κ isotype Ctrl antibody (RTK2758, Biolegend, San Diego, USA), PE Armenian Hamster IgG isotype Ctrl antibody (HTK888, Biolegend, San Diego, USA), PE/Cy7 Rat IgG1 κ isotype Ctrl antibody (RTK2071, Biolegend, San Diego, USA).

FACS:

Article Title: Antibody-secreting plasma cells persist for decades in human intestine
Article Snippet: .. Flow cytometry and FACS Single-cell suspensions from lamina propria were analyzed by flow cytometry according to standard procedure using fluorochrome-conjugated antibodies targeting HLA-A2–PE (Abcam); CD20-FITC, CD45-APC-H7, and Ki67-FITC (BD); Bcl2–Alexa Fluor 488, BCMA-PE, CD3-APC, CD14-APC, CD19–Alexa Flour 488/PE-Cy7, CD20-PE, CD27-BV421/Pacific blue/APC-Cy7/PE-Cy7, CD28–Alexa Fluor 488, CD38-PE/APC-Cy7, CD45-BV510/APC-Cy7, CD56–Alexa Flour 488, CD95-PE, and HLA-DR–BV605/PerCP (BioLegend); CD138-FITC/PE-Cy7 and HLA-A3–FITC/APC (eBioscience); HLA-B7–PE (EMD Millipore); HLA-B8–PE (Miltenyi Biotec); IgA-FITC/PE, IgM-FITC/PE, and IgD-FITC/PE (SouthernBiotech); and Blimp-1–DyLight 488 (Thermo Fisher Scientific). .. For detection of intracellular antigens, cells were fixed in formaldehyde and permeabilized using a Foxp3 staining buffer set (eBioscience).

Labeling:

Article Title: Divergent expression patterns of IL-4 and IL-13 define unique functions in allergic immunity
Article Snippet: Flow cytometric analysis of GATA-3 and Bcl-6 IL-4KN2/+ IL-13Smart/+ , IL-4KN2/4get , or IL-44get/+ IL-13Smart/+ mice were perfused with 20 ml PBS, and mediastinal lymph nodes and lung were collected. .. Single-cell suspensions were prepared and labeled with antibodies against surface molecules as listed: huCD4 (RPA-T4, PE-Cy7, FITC, or PE-anti-human CD4, eBioscience), huCD2, (PE or APC-α-CD2, Caltag), CD278 (C398.4A, APC-anti-ICOS, Biolegend), CD4 (RM4–5, APC-780-anti-CD4, eBioscience), CD8 (53-6.7, PerCP-Cy5.5-anti-CD8, BD Pharmingen), CD19 (ID3, Percp-Cy5.5-anti-CD19, BD Pharmingen), CD49b (DX5, PE-Cy7-anti-CD49b, eBioscience). .. After surface staining, cells were stained with Violet Live/Dead fixable stain (Invitrogen) to exclude dead cells.

Recombinase Polymerase Amplification:

Article Title: Divergent expression patterns of IL-4 and IL-13 define unique functions in allergic immunity
Article Snippet: Flow cytometric analysis of GATA-3 and Bcl-6 IL-4KN2/+ IL-13Smart/+ , IL-4KN2/4get , or IL-44get/+ IL-13Smart/+ mice were perfused with 20 ml PBS, and mediastinal lymph nodes and lung were collected. .. Single-cell suspensions were prepared and labeled with antibodies against surface molecules as listed: huCD4 (RPA-T4, PE-Cy7, FITC, or PE-anti-human CD4, eBioscience), huCD2, (PE or APC-α-CD2, Caltag), CD278 (C398.4A, APC-anti-ICOS, Biolegend), CD4 (RM4–5, APC-780-anti-CD4, eBioscience), CD8 (53-6.7, PerCP-Cy5.5-anti-CD8, BD Pharmingen), CD19 (ID3, Percp-Cy5.5-anti-CD19, BD Pharmingen), CD49b (DX5, PE-Cy7-anti-CD49b, eBioscience). .. After surface staining, cells were stained with Violet Live/Dead fixable stain (Invitrogen) to exclude dead cells.

Construct:

Article Title: Effective Depletion of Pre-existing Anti-AAV Antibodies Requires Broad Immune Targeting
Article Snippet: Flow Cytometry Splenocytes were assayed by flow cytometry to identify immune cell subsets and their status. .. Overlapping panels were constructed using antibodies against mouse CD3 (fluorescein isothiocyanate [FITC], 145-2C11, BD Pharmingen), CD4 (BV786, GK1.5 BD Horizon), CD19 (PerCP/Cy5.5, 6D5, BioLegend), CD62L (BV711, MEL-14, BioLegend), CD44 (PE, IM7, BioLegend), CD27 (BV421, M-T271, BioLegend), PD1 (Pe/Dazzle594, 29F.1A12, BioLegend), CXCR5 (biotin, 2G8, BD Biosciences), GL7 (Alexa647, Gl7, BioLegend), IgD (PeCy7, 11-26c.2a, BioLegend), and IgG (BV605, poly4053, BioLegend). .. For CXCR5 staining, cells were first stained with a biotinylated primary CXCR5 followed by streptavidin-APC (Molecular Probes).

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  • 99
    BioLegend pe labeled anti murine cd8
    Transfecting bystander cells leads to activation of antigen-specific naïve <t>CD8</t> + T cells. SIINFEKL-specific OT-1 T cells were co-cultured with DCs and transfected fibroblasts and the amount of IFN-γ secretion was determined by ELISA. Experiments
    Pe Labeled Anti Murine Cd8, supplied by BioLegend, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pe labeled anti murine cd8/product/BioLegend
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pe labeled anti murine cd8 - by Bioz Stars, 2021-07
    99/100 stars
      Buy from Supplier

    99
    BioLegend pe labeled anti ifn γ
    Immunomonitoring. A . Percentages of MDSCs by two criteria. B . Percentages of Tregs. C . Changes of tumor-reactive <t>IFN-γ</t> + cells (% of CD4 + or CD8 + T cells). Assay was performed as described in Methods section. D . The concentration of IL-8 in sera measured by a cytofluorometry-based ELISA system at different time points during treatment of the 8 patients.
    Pe Labeled Anti Ifn γ, supplied by BioLegend, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pe labeled anti ifn γ/product/BioLegend
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pe labeled anti ifn γ - by Bioz Stars, 2021-07
    99/100 stars
      Buy from Supplier

    99
    BioLegend pe labeled anti cd25
    Lung infection and analysis of blood mononuclear cells. A ) CT scan of the lungs of the patient, showing an area of consolidation of 43×60 mm in right lower lobe parenchyma (indicated by arrows). B ) Lung biopsy (sliced in two pieces) with visible cavitated lesions with mucoid material. C ) Hematoxylin/eosin staining of the lung biopsy showing the infiltration of inflammatory cells with predominance of histiocytes and multinucleated giant cells containing PAS-positive spherical bodies of 5 to 10 μm in diameter (indicated by arrows). The morphological characteristics of these PAS-positive spherical bodies are compatible with Cryptococcus neoformans infection. D ) Absolute numbers of total NK cells, CD3 − CD56 dim cells and CD3 − CD56 bright cells in blood of 6 healthy normal donors (N) and in blood from two draws from different dates (indicated in parentheses) from the patient (P). E ) PBMCs from 9 different normal donors (N) and from different blood draws from different dates (indicated in parentheses) from the patient (P) were stained with anti-CD3 and anti-CD56 mAbs, and the percentage of NK cells (CD3 − CD56 + cells), CD3 − CD56 dim and CD3 − CD56 bright cells within the whole mononuclear cell population (PBMCs) were calculated and depicted as dot plots. Also, the relative abundance of CD3 − CD56 bright and CD3 − CD56 dim cells was calculated and depicted (CD3 − CD56 bright are shown in black bars; CD3 − CD56 dim cells are shown in white bars). Each sample of the patient was tested at least twice, and individual values obtained are shown as a dot in the graphs. Interquartile ranges (IQR) are indicated in the graphs of panels D and E. F ) Representative dot plots of lymphoid cells gated according to their FSC and SSC parameters. The numbers in each region correspond to the percentage of CD3 − CD56 dim and CD3 − CD56 bright cells within the lymphoid population (gated according to their FSC and SSC parameters). G ) Representative dot plots to show the percentage of CD3 − CD56 − CD16 + NK cells within the CD3 − lymphoid cell population in a healthy normal control (N) and in one sample of the patient (P). The numbers in each quadrant correspond to the percentage of CD16 + CD56 − , CD16 + CD56 dim and CD16 − CD56 bright NK cells. Right graph: data from 3 healthy normal controls and 3 blood samples from different dates from the patient (P). Percentages depicted correspond to the percentage of CD3 − CD56 − CD16 + cells within the total NK cell population (CD16 + CD56 − + CD16 + CD56 dim + CD16 − CD56 bright cells). H ) PBMCs from normal donors (N) and from the patient (P) were stained with anti-CD3, anti-CD14 and anti-HLA class I mAbs to assess HLA class I expression (continuous line) on T cells (CD3 + cells) and monocytes (CD14 + cells). Data presented correspond to representative histograms. Dashed lines: IC mAb. The numbers inserted in the graphs correspond to the MFI. I ) PBMCs from normal donors (N) and from the patient (P) were stained with anti-CD4, <t>anti-CD25</t> and anti-Foxp3 to assess the percentage of Tregs. The dot plots correspond to Foxp3 vs . CD25 in CD4 + cells gated from the FSC vs. SSC plots. The percentages in each quadrant are shown. The right graph shows the percentage of Tregs in PBMCs from 6 healthy normal controls and in two blood samples from different dates (indicated in parentheses) from the patient (P). One representative experiment from 3 independent analyses is shown in C and D. PBMCs from blood samples obtained in February 2011 were used for panels F, H and I.
    Pe Labeled Anti Cd25, supplied by BioLegend, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pe labeled anti cd25/product/BioLegend
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pe labeled anti cd25 - by Bioz Stars, 2021-07
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      Buy from Supplier

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    Transfecting bystander cells leads to activation of antigen-specific naïve CD8 + T cells. SIINFEKL-specific OT-1 T cells were co-cultured with DCs and transfected fibroblasts and the amount of IFN-γ secretion was determined by ELISA. Experiments

    Journal: Journal of Controlled Release

    Article Title: Polymer-Mediated DNA Vaccine Delivery via Bystander Cells Requires a Proper Balance between Transfection Efficiency and Cytotoxicity

    doi: 10.1016/j.jconrel.2011.08.037

    Figure Lengend Snippet: Transfecting bystander cells leads to activation of antigen-specific naïve CD8 + T cells. SIINFEKL-specific OT-1 T cells were co-cultured with DCs and transfected fibroblasts and the amount of IFN-γ secretion was determined by ELISA. Experiments

    Article Snippet: An aliquot of cells was stained with PE-labeled anti-murine CD8 (BioLegend) and PerCP Cy 5.5-labeled anti-murine CD90 (CD90.1) (BioLegend).

    Techniques: Activation Assay, Cell Culture, Transfection, Enzyme-linked Immunosorbent Assay

    Schematic illustration of a co-culture system for the study of polymer-mediated DNA vaccine delivery to bystander cells. The process involves bystander cells (fibroblasts), dendritic cells (DCs), and antigen-specific CD8 + T cells, mixed sequentially and

    Journal: Journal of Controlled Release

    Article Title: Polymer-Mediated DNA Vaccine Delivery via Bystander Cells Requires a Proper Balance between Transfection Efficiency and Cytotoxicity

    doi: 10.1016/j.jconrel.2011.08.037

    Figure Lengend Snippet: Schematic illustration of a co-culture system for the study of polymer-mediated DNA vaccine delivery to bystander cells. The process involves bystander cells (fibroblasts), dendritic cells (DCs), and antigen-specific CD8 + T cells, mixed sequentially and

    Article Snippet: An aliquot of cells was stained with PE-labeled anti-murine CD8 (BioLegend) and PerCP Cy 5.5-labeled anti-murine CD90 (CD90.1) (BioLegend).

    Techniques: Co-Culture Assay

    Immunomonitoring. A . Percentages of MDSCs by two criteria. B . Percentages of Tregs. C . Changes of tumor-reactive IFN-γ + cells (% of CD4 + or CD8 + T cells). Assay was performed as described in Methods section. D . The concentration of IL-8 in sera measured by a cytofluorometry-based ELISA system at different time points during treatment of the 8 patients.

    Journal: Journal for Immunotherapy of Cancer

    Article Title: A pilot study of autologous tumor lysate-loaded dendritic cell vaccination combined with sunitinib for metastatic renal cell carcinoma

    doi: 10.1186/s40425-014-0030-4

    Figure Lengend Snippet: Immunomonitoring. A . Percentages of MDSCs by two criteria. B . Percentages of Tregs. C . Changes of tumor-reactive IFN-γ + cells (% of CD4 + or CD8 + T cells). Assay was performed as described in Methods section. D . The concentration of IL-8 in sera measured by a cytofluorometry-based ELISA system at different time points during treatment of the 8 patients.

    Article Snippet: After incubation for 45 min, the cells were washed with cold PBS and treated with Fixable viability dye eFluor 450 (eBioscience, San Diego, CA), PE-labeled anti-IFN-γ (detection reagent), Alexa Fluor 647-labeled anti-human CD3 (Biolegend, San Diego, CA), PC5-labeled anti-human CD8 (Beckman Coulter, Fullerton, CA), and PECy7-labeled anti-human CD4 (Biolegend) mAbs.

    Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay

    Activation of α7 nAChR decreases NK cell-mediated cytotoxicity and IFN-γ production. A , expression of MICA in MB49 cells. MB49 were transduced with a retroviral vector encoding MICA*008 ( left panel ) or with empty vector ( right panel ).

    Journal: The Journal of Biological Chemistry

    Article Title: Expression and Functional Role of α7 Nicotinic Receptor in Human Cytokine-stimulated Natural Killer (NK) Cells *

    doi: 10.1074/jbc.M115.710574

    Figure Lengend Snippet: Activation of α7 nAChR decreases NK cell-mediated cytotoxicity and IFN-γ production. A , expression of MICA in MB49 cells. MB49 were transduced with a retroviral vector encoding MICA*008 ( left panel ) or with empty vector ( right panel ).

    Article Snippet: The following monoclonal antibodies (mAbs) against human molecules were used: FITC- and PE-labeled anti-CD3 (UCHT-1, Southern Biotech); PE/Cy5- and PE/Cy7-labeled anti-CD56 (N901, Beckman Coulter); PE-labeled anti-IFN-γ (4S.B3, Biolegend); PE-labeled anti-NKG2D mAb (clone 1D11, Biolegend); PE-labeled anti-NKp46 (clone 9E2; Biolegend); FITC-labeled anti-CD226 (DNAM-1, clone DX11, BD Pharmingen); FITC anti-CD14 (clone HCD14, Biolegend); FITC anti-HLA-DR (clone L243, Biolegend); PE anti-CD86 (clone IT2.2, Biolegend); FITC anti-CD83 (clone HB15e, Biolegend); Alexa 488-labeled anti-MICA (clone 159227, R & D Systems); isotype-matched negative control mAb (IC mAb, eBioscience). α-BTX and Alexa Fluor 488-labeled α-BTX were from Molecular Probes; PNU-120596 and PNU-282987 were from Santa Cruz Biotechnology; EGTA was from Sigma; and BAPTA/AM was from Calbiochem (San Diego, CA). pMSCV and pMSCV/MICA*008 retroviral DNA (negative control or encoding MICA*008, respectively) were kindly provided by Dr. Alessandra Zingoni and Dr. Angela Santoni from the Laboratory of Molecular Immunology and Immunopathology, Department of Molecular Medicine, Sapienza University of Rome, Italy.

    Techniques: Activation Assay, Expressing, Transduction, Plasmid Preparation

    Lung infection and analysis of blood mononuclear cells. A ) CT scan of the lungs of the patient, showing an area of consolidation of 43×60 mm in right lower lobe parenchyma (indicated by arrows). B ) Lung biopsy (sliced in two pieces) with visible cavitated lesions with mucoid material. C ) Hematoxylin/eosin staining of the lung biopsy showing the infiltration of inflammatory cells with predominance of histiocytes and multinucleated giant cells containing PAS-positive spherical bodies of 5 to 10 μm in diameter (indicated by arrows). The morphological characteristics of these PAS-positive spherical bodies are compatible with Cryptococcus neoformans infection. D ) Absolute numbers of total NK cells, CD3 − CD56 dim cells and CD3 − CD56 bright cells in blood of 6 healthy normal donors (N) and in blood from two draws from different dates (indicated in parentheses) from the patient (P). E ) PBMCs from 9 different normal donors (N) and from different blood draws from different dates (indicated in parentheses) from the patient (P) were stained with anti-CD3 and anti-CD56 mAbs, and the percentage of NK cells (CD3 − CD56 + cells), CD3 − CD56 dim and CD3 − CD56 bright cells within the whole mononuclear cell population (PBMCs) were calculated and depicted as dot plots. Also, the relative abundance of CD3 − CD56 bright and CD3 − CD56 dim cells was calculated and depicted (CD3 − CD56 bright are shown in black bars; CD3 − CD56 dim cells are shown in white bars). Each sample of the patient was tested at least twice, and individual values obtained are shown as a dot in the graphs. Interquartile ranges (IQR) are indicated in the graphs of panels D and E. F ) Representative dot plots of lymphoid cells gated according to their FSC and SSC parameters. The numbers in each region correspond to the percentage of CD3 − CD56 dim and CD3 − CD56 bright cells within the lymphoid population (gated according to their FSC and SSC parameters). G ) Representative dot plots to show the percentage of CD3 − CD56 − CD16 + NK cells within the CD3 − lymphoid cell population in a healthy normal control (N) and in one sample of the patient (P). The numbers in each quadrant correspond to the percentage of CD16 + CD56 − , CD16 + CD56 dim and CD16 − CD56 bright NK cells. Right graph: data from 3 healthy normal controls and 3 blood samples from different dates from the patient (P). Percentages depicted correspond to the percentage of CD3 − CD56 − CD16 + cells within the total NK cell population (CD16 + CD56 − + CD16 + CD56 dim + CD16 − CD56 bright cells). H ) PBMCs from normal donors (N) and from the patient (P) were stained with anti-CD3, anti-CD14 and anti-HLA class I mAbs to assess HLA class I expression (continuous line) on T cells (CD3 + cells) and monocytes (CD14 + cells). Data presented correspond to representative histograms. Dashed lines: IC mAb. The numbers inserted in the graphs correspond to the MFI. I ) PBMCs from normal donors (N) and from the patient (P) were stained with anti-CD4, anti-CD25 and anti-Foxp3 to assess the percentage of Tregs. The dot plots correspond to Foxp3 vs . CD25 in CD4 + cells gated from the FSC vs. SSC plots. The percentages in each quadrant are shown. The right graph shows the percentage of Tregs in PBMCs from 6 healthy normal controls and in two blood samples from different dates (indicated in parentheses) from the patient (P). One representative experiment from 3 independent analyses is shown in C and D. PBMCs from blood samples obtained in February 2011 were used for panels F, H and I.

    Journal: PLoS ONE

    Article Title: Human Natural Killer Cell Maturation Defect Supports In Vivo CD56bright to CD56dim Lineage Development

    doi: 10.1371/journal.pone.0051677

    Figure Lengend Snippet: Lung infection and analysis of blood mononuclear cells. A ) CT scan of the lungs of the patient, showing an area of consolidation of 43×60 mm in right lower lobe parenchyma (indicated by arrows). B ) Lung biopsy (sliced in two pieces) with visible cavitated lesions with mucoid material. C ) Hematoxylin/eosin staining of the lung biopsy showing the infiltration of inflammatory cells with predominance of histiocytes and multinucleated giant cells containing PAS-positive spherical bodies of 5 to 10 μm in diameter (indicated by arrows). The morphological characteristics of these PAS-positive spherical bodies are compatible with Cryptococcus neoformans infection. D ) Absolute numbers of total NK cells, CD3 − CD56 dim cells and CD3 − CD56 bright cells in blood of 6 healthy normal donors (N) and in blood from two draws from different dates (indicated in parentheses) from the patient (P). E ) PBMCs from 9 different normal donors (N) and from different blood draws from different dates (indicated in parentheses) from the patient (P) were stained with anti-CD3 and anti-CD56 mAbs, and the percentage of NK cells (CD3 − CD56 + cells), CD3 − CD56 dim and CD3 − CD56 bright cells within the whole mononuclear cell population (PBMCs) were calculated and depicted as dot plots. Also, the relative abundance of CD3 − CD56 bright and CD3 − CD56 dim cells was calculated and depicted (CD3 − CD56 bright are shown in black bars; CD3 − CD56 dim cells are shown in white bars). Each sample of the patient was tested at least twice, and individual values obtained are shown as a dot in the graphs. Interquartile ranges (IQR) are indicated in the graphs of panels D and E. F ) Representative dot plots of lymphoid cells gated according to their FSC and SSC parameters. The numbers in each region correspond to the percentage of CD3 − CD56 dim and CD3 − CD56 bright cells within the lymphoid population (gated according to their FSC and SSC parameters). G ) Representative dot plots to show the percentage of CD3 − CD56 − CD16 + NK cells within the CD3 − lymphoid cell population in a healthy normal control (N) and in one sample of the patient (P). The numbers in each quadrant correspond to the percentage of CD16 + CD56 − , CD16 + CD56 dim and CD16 − CD56 bright NK cells. Right graph: data from 3 healthy normal controls and 3 blood samples from different dates from the patient (P). Percentages depicted correspond to the percentage of CD3 − CD56 − CD16 + cells within the total NK cell population (CD16 + CD56 − + CD16 + CD56 dim + CD16 − CD56 bright cells). H ) PBMCs from normal donors (N) and from the patient (P) were stained with anti-CD3, anti-CD14 and anti-HLA class I mAbs to assess HLA class I expression (continuous line) on T cells (CD3 + cells) and monocytes (CD14 + cells). Data presented correspond to representative histograms. Dashed lines: IC mAb. The numbers inserted in the graphs correspond to the MFI. I ) PBMCs from normal donors (N) and from the patient (P) were stained with anti-CD4, anti-CD25 and anti-Foxp3 to assess the percentage of Tregs. The dot plots correspond to Foxp3 vs . CD25 in CD4 + cells gated from the FSC vs. SSC plots. The percentages in each quadrant are shown. The right graph shows the percentage of Tregs in PBMCs from 6 healthy normal controls and in two blood samples from different dates (indicated in parentheses) from the patient (P). One representative experiment from 3 independent analyses is shown in C and D. PBMCs from blood samples obtained in February 2011 were used for panels F, H and I.

    Article Snippet: The following mAbs against human molecules were used: FITC-, PE- and SPRD-labeled anti-CD3 (UCHT-1, Southern Biotech); PE/Cy7-labeled anti-CD3 (UCHT-1, Biolegend); FITC-labeled anti-CD14 (HCD14 Biolegend); PE-labeled anti-HLA class I (W6/32, Biolegend); PE/Cy5-labeled anti-CD56 (N901, Beckman Coulter); FITC-labeled anti-CD16 (eBioCB16, eBioscience); FITC-labeled anti-CD57 (HCD57, Biolegend); FITC- or PE-labeled anti-CD62L (DREG-56, Southern Biotech); FITC-labeled anti-perforin (pfp, dG9, Biolegend); PE-labeled anti-IFN-γ (4S.B3, Biolegend); FITC-labeled anti-CD107a (1D4B, BD Biosciences); PE-labeled anti-CD25 (BC96, Biolegend); FITC-labeled anti-CD69 (FN50, BD); PE-labeled anti-NKG2D mAb (clone 1D11, Biolegend); PE-labeled anti-NKp46 (clone 9E2; Biolegend); FITC-labeled anti-CD226 (DNAM-1, clone DX11, BD Pharmingen); PE-labeled anti-CD244 (2B4, clone C1.7, Biolegend); PE-labeled anti CD94 (clone DX22; Biolegend); APC-labeled anti-NKG2A (clone 131411, R & D); mAb isotype-matched control mAb (IC, eBioscience).

    Techniques: Infection, Computed Tomography, Staining, Expressing