anti pd 1 mab  (Valiant)

 
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    Name:
    Anti Desmoplakin 1 2 mouse monoclonal antibody
    Description:
    The anti desmoplakin 1 2 is a mouse IgG1 immunoglobulin It is purified by affinity chromatography The antigen is bovine desmoplakins 1 2
    Catalog Number:
    08695421
    Price:
    718.05
    Category:
    Life Sciences Antibodies Primary Antibodies
    Applications:
    Immunofluorescence assays (IFA) ,Immunohistochemistry, Immunoblot
    Size:
    50 µL
    Buy from Supplier


    Structured Review

    Valiant anti pd 1 mab
    Effect of <t>PD-1</t> blockade on T-cell function. (a) PBMC from BLV + cattle were cultured with Flag peptide as negative control and gp51 peptide mix. IFN-γ production was measured by ELISA ( n = 13). (b, c, d) PBMC were cultured with rat IgG control or anti-PD-1 mAb (5D2; 20 μg/mL) in the presence of gp51 peptide mix. IFN-γ and IL-10 production was measured by ELISA ( b ; n = 22, d; n = 26). Positive correlation between increasing rate of IFN-γ production and percentages of PD-1 + cells in CD4 + T cells corresponding to Figure 4 a ( c ; n = 22). Correlation statistics were analyzed using the Spearman correlation. (e) The proliferative responses were evaluated by detection of CFSE lo cells in IgM - lymphocytes by flow cytometry ( n = 12). Representative dot plots of CFSE-staining in lymphocytes other than B cells are shown. Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P
    The anti desmoplakin 1 2 is a mouse IgG1 immunoglobulin It is purified by affinity chromatography The antigen is bovine desmoplakins 1 2
    https://www.bioz.com/result/anti pd 1 mab/product/Valiant
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti pd 1 mab - by Bioz Stars, 2021-04
    93/100 stars

    Images

    1) Product Images from "Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vitro"

    Article Title: Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vitro

    Journal: Veterinary Research

    doi: 10.1186/1297-9716-44-59

    Effect of PD-1 blockade on T-cell function. (a) PBMC from BLV + cattle were cultured with Flag peptide as negative control and gp51 peptide mix. IFN-γ production was measured by ELISA ( n = 13). (b, c, d) PBMC were cultured with rat IgG control or anti-PD-1 mAb (5D2; 20 μg/mL) in the presence of gp51 peptide mix. IFN-γ and IL-10 production was measured by ELISA ( b ; n = 22, d; n = 26). Positive correlation between increasing rate of IFN-γ production and percentages of PD-1 + cells in CD4 + T cells corresponding to Figure 4 a ( c ; n = 22). Correlation statistics were analyzed using the Spearman correlation. (e) The proliferative responses were evaluated by detection of CFSE lo cells in IgM - lymphocytes by flow cytometry ( n = 12). Representative dot plots of CFSE-staining in lymphocytes other than B cells are shown. Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P
    Figure Legend Snippet: Effect of PD-1 blockade on T-cell function. (a) PBMC from BLV + cattle were cultured with Flag peptide as negative control and gp51 peptide mix. IFN-γ production was measured by ELISA ( n = 13). (b, c, d) PBMC were cultured with rat IgG control or anti-PD-1 mAb (5D2; 20 μg/mL) in the presence of gp51 peptide mix. IFN-γ and IL-10 production was measured by ELISA ( b ; n = 22, d; n = 26). Positive correlation between increasing rate of IFN-γ production and percentages of PD-1 + cells in CD4 + T cells corresponding to Figure 4 a ( c ; n = 22). Correlation statistics were analyzed using the Spearman correlation. (e) The proliferative responses were evaluated by detection of CFSE lo cells in IgM - lymphocytes by flow cytometry ( n = 12). Representative dot plots of CFSE-staining in lymphocytes other than B cells are shown. Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P

    Techniques Used: Cell Function Assay, Cell Culture, Negative Control, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Cytometry, Staining

    Recognition of PD-1-expressing lymphocytes by anti-PD-1 mAb. (a) Gating strategies for CD4 + T cells, CD8 + T cells and IgM + B cells in bovine lymphocytes. (b) Representative histograms obtained by flow cytometry of PD-1 expression in CD4 + T cells, CD8 + T cells and IgM + B cells isolated from three BLV - cattle. Freshly isolated PBMC were stained with anti-PD-1 (5D2), CD4, CD8, and IgM Ab. PBMC were cultivated with PBS (No stimulation), PMA/ionomycin, and PWM for 24, 48, and 72 h, and stained in a similar way. (c) Proportions of PD-1 positive cells in CD4 + T cells, CD8 + T cells and IgM + B cells. Statistical comparisons between percentages of PD-1 positive cells stimulated with PBS and PMA/ionomycin or PWM were made using two way ANOVA. Differences were considered statistically significant at P
    Figure Legend Snippet: Recognition of PD-1-expressing lymphocytes by anti-PD-1 mAb. (a) Gating strategies for CD4 + T cells, CD8 + T cells and IgM + B cells in bovine lymphocytes. (b) Representative histograms obtained by flow cytometry of PD-1 expression in CD4 + T cells, CD8 + T cells and IgM + B cells isolated from three BLV - cattle. Freshly isolated PBMC were stained with anti-PD-1 (5D2), CD4, CD8, and IgM Ab. PBMC were cultivated with PBS (No stimulation), PMA/ionomycin, and PWM for 24, 48, and 72 h, and stained in a similar way. (c) Proportions of PD-1 positive cells in CD4 + T cells, CD8 + T cells and IgM + B cells. Statistical comparisons between percentages of PD-1 positive cells stimulated with PBS and PMA/ionomycin or PWM were made using two way ANOVA. Differences were considered statistically significant at P

    Techniques Used: Expressing, Flow Cytometry, Cytometry, Isolation, Staining

    Enhancement of cytokine production by anti-PD-1 mAb in bovine lymphocytes. PBMC were cultivated with rat IgG control or four types of anti-PD-1 mAb (20 μg/mL) in the absence ( a ; n = 8 or 14) or presence ( b ; n = 16) of PMA/ionomycin. IFN-γ production was measured by ELISA. Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P
    Figure Legend Snippet: Enhancement of cytokine production by anti-PD-1 mAb in bovine lymphocytes. PBMC were cultivated with rat IgG control or four types of anti-PD-1 mAb (20 μg/mL) in the absence ( a ; n = 8 or 14) or presence ( b ; n = 16) of PMA/ionomycin. IFN-γ production was measured by ELISA. Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Expression analysis of bovine PD-1 in BLV-infected cattle. (a, b) Percentages of PD-1-expressing CD4 + and CD8 + T cells in PBMC (a) and LN (b) isolated from BLV - cattle (CD4 + T cells; n = 20 and 31, CD8 + T cells; n = 16 and 31), BLV + cattle (CD4 + T cells; n = 35 and 15, CD8 + T cells; n = 28 and 15) and BCBL (CD4 + T cells; n = 7 and 6, CD8 + T cells; n = 7 and 6). (c) An example of gating strategies for PD-1 expression in CD4 + T cells and CD8 + T cells isolated from lymph node of BCBL and BLV - cattle. Values in the quadrant indicate the percentage of the cells in lymphocytes other than IgM + B cells. (d) Percentages of PD-L1-expressing gp51 + ( n = 5) and gp51 - ( n = 5) in IgM + B cells from BLV + and BLV - cattle ( n = 9). Representative dot plots of PD-L1 staining in gp51 + or gp51 - B cells are also shown. Statistical comparisons were made using one-way ANOVA with Tukey’s test. Differences were considered statistically significant at P
    Figure Legend Snippet: Expression analysis of bovine PD-1 in BLV-infected cattle. (a, b) Percentages of PD-1-expressing CD4 + and CD8 + T cells in PBMC (a) and LN (b) isolated from BLV - cattle (CD4 + T cells; n = 20 and 31, CD8 + T cells; n = 16 and 31), BLV + cattle (CD4 + T cells; n = 35 and 15, CD8 + T cells; n = 28 and 15) and BCBL (CD4 + T cells; n = 7 and 6, CD8 + T cells; n = 7 and 6). (c) An example of gating strategies for PD-1 expression in CD4 + T cells and CD8 + T cells isolated from lymph node of BCBL and BLV - cattle. Values in the quadrant indicate the percentage of the cells in lymphocytes other than IgM + B cells. (d) Percentages of PD-L1-expressing gp51 + ( n = 5) and gp51 - ( n = 5) in IgM + B cells from BLV + and BLV - cattle ( n = 9). Representative dot plots of PD-L1 staining in gp51 + or gp51 - B cells are also shown. Statistical comparisons were made using one-way ANOVA with Tukey’s test. Differences were considered statistically significant at P

    Techniques Used: Expressing, Infection, Isolation, Staining

    Effect of PD-1 blockade on gp51 expression and B-cell activation. (a , b, c) Percentages of gp51-expressing cells ( a ; n = 15) and mean fluorescence index (MFI) of WC4 (CD19 like molecule) ( b ; n = 12) and CD80 ( c ; n = 6) in IgM + B cells were evaluated by flow cytometry in PBMC treated with rat IgG control or anti-PD-1 mAb (20 μg/mL). Representative contour plots showing gp51 expression (right panels) in PBMC treated with rat IgG control (upper panels) or anti-PD-1 mAb (lower panels) is shown in (a) . No staining was observed in PBMC stained with isotype control for anti-gp51 mAb (left panels). (d) Expression of BAFF mRNA was evaluated by real-time PCR ( n = 12). The results are indicated as relative change to control (no antibody treatment) when the amount of BAFF mRNA expression is divided by GAPDH mRNA expression. (e) Percentages of apoptotic cells in IgM + B cells were measured by flow cytometry. Apoptotic B cells were identified as annexin-V + 7-AAD - cells ( n = 11). (f) Percentages of gp51-expressing cells were evaluated by flow cytometry in isolated B cells cultivated with rat IgG control or anti-PD-1 mAb ( n = 11). Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P
    Figure Legend Snippet: Effect of PD-1 blockade on gp51 expression and B-cell activation. (a , b, c) Percentages of gp51-expressing cells ( a ; n = 15) and mean fluorescence index (MFI) of WC4 (CD19 like molecule) ( b ; n = 12) and CD80 ( c ; n = 6) in IgM + B cells were evaluated by flow cytometry in PBMC treated with rat IgG control or anti-PD-1 mAb (20 μg/mL). Representative contour plots showing gp51 expression (right panels) in PBMC treated with rat IgG control (upper panels) or anti-PD-1 mAb (lower panels) is shown in (a) . No staining was observed in PBMC stained with isotype control for anti-gp51 mAb (left panels). (d) Expression of BAFF mRNA was evaluated by real-time PCR ( n = 12). The results are indicated as relative change to control (no antibody treatment) when the amount of BAFF mRNA expression is divided by GAPDH mRNA expression. (e) Percentages of apoptotic cells in IgM + B cells were measured by flow cytometry. Apoptotic B cells were identified as annexin-V + 7-AAD - cells ( n = 11). (f) Percentages of gp51-expressing cells were evaluated by flow cytometry in isolated B cells cultivated with rat IgG control or anti-PD-1 mAb ( n = 11). Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P

    Techniques Used: Expressing, Activation Assay, Fluorescence, Flow Cytometry, Cytometry, Staining, Real-time Polymerase Chain Reaction, Isolation

    Recognition of PD-1-expressing cells by anti-PD-1 mAb. (a) Flow cytometric analysis of surface expression of bovine PD-1. Cos-7 expressing PD-1 (black line) and Cos-7 transfected with the control vector (dashed line) were stained with four types of anti-PD-1 mAb (2C12, 2H7, 3G2, and 5D2) and isotype control (Rat IgG). (b) Western blotting analysis of bovine PD-1 expression in CHO-DG44 cells stably expressing bovine PD-1. Three types of anti-PD-1 mAb recognized PD-1 (triangle) at about 68 kDa. Anti-actin antibody and anti-myc antibody was used as a loading control and a positive control.
    Figure Legend Snippet: Recognition of PD-1-expressing cells by anti-PD-1 mAb. (a) Flow cytometric analysis of surface expression of bovine PD-1. Cos-7 expressing PD-1 (black line) and Cos-7 transfected with the control vector (dashed line) were stained with four types of anti-PD-1 mAb (2C12, 2H7, 3G2, and 5D2) and isotype control (Rat IgG). (b) Western blotting analysis of bovine PD-1 expression in CHO-DG44 cells stably expressing bovine PD-1. Three types of anti-PD-1 mAb recognized PD-1 (triangle) at about 68 kDa. Anti-actin antibody and anti-myc antibody was used as a loading control and a positive control.

    Techniques Used: Expressing, Flow Cytometry, Transfection, Plasmid Preparation, Staining, Western Blot, Stable Transfection, Positive Control

    Related Articles

    Western Blot:

    Article Title: c-FLIP Mediates Resistance of Hodgkin/Reed-Sternberg Cells to Death Receptor-induced Apoptosis
    Article Snippet: .. For Western blot analysis the following primary antibodies were used: monoclonal anti–c-FLIPS/L (Dave-2; Qbiogene and G-11; Santa Cruz Biotechnology, Inc.), polyclonal anti-IκBα (C-21), polyclonal anti-CD95 (C-20), polyclonal anti-p65 (A; all from Santa Cruz Biotechnology, Inc.), monoclonal anti-FADD (Clone-1 and Clone A66-2; BD Biosciences), monoclonal anti–tubulin α (MCA78A; Serotec), monoclonal anti–C-8 (C-15; provided by P. Krammer), and polyclonal anti–Bcl-xL (Transduction Laboratories). .. Filters were incubated with horseradish peroxidase–conjugated secondary antibodies.

    Transduction:

    Article Title: c-FLIP Mediates Resistance of Hodgkin/Reed-Sternberg Cells to Death Receptor-induced Apoptosis
    Article Snippet: .. For Western blot analysis the following primary antibodies were used: monoclonal anti–c-FLIPS/L (Dave-2; Qbiogene and G-11; Santa Cruz Biotechnology, Inc.), polyclonal anti-IκBα (C-21), polyclonal anti-CD95 (C-20), polyclonal anti-p65 (A; all from Santa Cruz Biotechnology, Inc.), monoclonal anti-FADD (Clone-1 and Clone A66-2; BD Biosciences), monoclonal anti–tubulin α (MCA78A; Serotec), monoclonal anti–C-8 (C-15; provided by P. Krammer), and polyclonal anti–Bcl-xL (Transduction Laboratories). .. Filters were incubated with horseradish peroxidase–conjugated secondary antibodies.

    Incubation:

    Article Title: Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vitro
    Article Snippet: The samples were separated on 12% SDS-polyacrylamide gels and transferred onto a polyvinylidene difluoride membrane (Merck Millipore, MA, USA). .. After being blocked with 3% skim milk in phosphate-buffered saline (PBS, pH 7.2) containing 0.05% Tween 20 (PBS-T), the membranes were incubated at room temperature for 2 h with anti-PD-1 mAb (2C12 and 3G2: 3 μg/mL, 2H7 and 5D2: 1 μg/mL), followed by washing and incubation with horse radish peroxidase (HRP)-conjugated anti-rat IgG (MP Biomedicals, CA, USA). .. The membrane was also probed with anti-Actin antibody (Merck Millipore; clone C4) and anti-myc tag antibody (Abcam, Cambridge, UK; goat polyclonal antibody) as loading control and positive control.

    Purification:

    Article Title: The delivery site of a monovalent influenza vaccine within the respiratory tract impacts on the immune response
    Article Snippet: Total antibody ELISA Total IgG and IgA titres in BALs or nasal washes were determined by ELISA as above, substituting influenza HA with goat anti-mouse IgG (Jackson ImmunoResearch) or goat anti-mouse IgA (Sigma). .. Serial dilutions of BALs or nasal washes, and standard curves constituted from mouse IgG (Chrompure, Jackson ImmunoResearch) or purified mouse myeloma protein IgA (MP Biomedicals, Aurora, OH) were added to the plates. ..

    Activated Clotting Time Assay:

    Article Title: RacGAP50C directs perinuclear ?-tubulin localization to organize the uniform microtubule array required for Drosophila myotube extension
    Article Snippet: For Phalloidin labeling, embryos were fixed for 6 minutes in 33% formaldehyde and 10% EGTA, and de-vitellinized by hand. .. Antibodies were used as follows: mouse anti-Muscle myosin [1:20, MAb FMM5C8 ( )], rabbit anti-β-galactosidase (1:1000, MP Biomedicals), mouse anti-αPS2 integrin, mouse anti-Even skipped and mouse anti-α-Spectrin (1:10) (all from the Developmental Studies Hybridoma Bank), guinea pig anti-Runt (1:2000, preabsorbed, gift from J. Reinitz, Stony Brook University, Stony Brook, NY, USA), rabbit anti-Vestigial (1:50, gift from S. Carroll, HHMI, University of Wisconsin, Madison, WI, USA), rabbit anti-RacGAP50C (1:500, gift from D. Glover, University of Cambridge, Cambridge, UK), rat anti-RacGAP50C (1:500, gift from R. Saint, Australian National University, Canberra, ACT, Australia), rabbit anti-β3-tubulin (1:5000, gift from D. Buttgereit, Philipps-Universitat Marburg, Marburg, Germany), mouse anti-Roundabout (1:10), mouse anti-γ-tubulin (1:500, Sigma, clone GTU-88), rabbit anti-Mef2 (1:2000, gift from B. Paterson, National Cancer Institute, NIH, Bethesda, MD, USA), Alexa Fluor 555-conjugated rabbit anti-GFP (1:250), Alexa Fluor 488-conjugated Phalloidin (1:500), Alexa Fluor 488-conjugated goat anti-rabbit, -mouse and -guinea pig (1:500), Alexa Fluor 555-conjugated goat anti-rabbit and -mouse (1:500) (all Alexa Fluors from Molecular Probes), and Cy3-conjugated goat anti-mouse (1:500, Jackson Laboratories). ..

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    Valiant anti pd 1 mab
    Effect of <t>PD-1</t> blockade on T-cell function. (a) PBMC from BLV + cattle were cultured with Flag peptide as negative control and gp51 peptide mix. IFN-γ production was measured by ELISA ( n = 13). (b, c, d) PBMC were cultured with rat IgG control or anti-PD-1 mAb (5D2; 20 μg/mL) in the presence of gp51 peptide mix. IFN-γ and IL-10 production was measured by ELISA ( b ; n = 22, d; n = 26). Positive correlation between increasing rate of IFN-γ production and percentages of PD-1 + cells in CD4 + T cells corresponding to Figure 4 a ( c ; n = 22). Correlation statistics were analyzed using the Spearman correlation. (e) The proliferative responses were evaluated by detection of CFSE lo cells in IgM - lymphocytes by flow cytometry ( n = 12). Representative dot plots of CFSE-staining in lymphocytes other than B cells are shown. Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P
    Anti Pd 1 Mab, supplied by Valiant, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti pd 1 mab/product/Valiant
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti pd 1 mab - by Bioz Stars, 2021-04
    93/100 stars
      Buy from Supplier

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    Effect of PD-1 blockade on T-cell function. (a) PBMC from BLV + cattle were cultured with Flag peptide as negative control and gp51 peptide mix. IFN-γ production was measured by ELISA ( n = 13). (b, c, d) PBMC were cultured with rat IgG control or anti-PD-1 mAb (5D2; 20 μg/mL) in the presence of gp51 peptide mix. IFN-γ and IL-10 production was measured by ELISA ( b ; n = 22, d; n = 26). Positive correlation between increasing rate of IFN-γ production and percentages of PD-1 + cells in CD4 + T cells corresponding to Figure 4 a ( c ; n = 22). Correlation statistics were analyzed using the Spearman correlation. (e) The proliferative responses were evaluated by detection of CFSE lo cells in IgM - lymphocytes by flow cytometry ( n = 12). Representative dot plots of CFSE-staining in lymphocytes other than B cells are shown. Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P

    Journal: Veterinary Research

    Article Title: Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vitro

    doi: 10.1186/1297-9716-44-59

    Figure Lengend Snippet: Effect of PD-1 blockade on T-cell function. (a) PBMC from BLV + cattle were cultured with Flag peptide as negative control and gp51 peptide mix. IFN-γ production was measured by ELISA ( n = 13). (b, c, d) PBMC were cultured with rat IgG control or anti-PD-1 mAb (5D2; 20 μg/mL) in the presence of gp51 peptide mix. IFN-γ and IL-10 production was measured by ELISA ( b ; n = 22, d; n = 26). Positive correlation between increasing rate of IFN-γ production and percentages of PD-1 + cells in CD4 + T cells corresponding to Figure 4 a ( c ; n = 22). Correlation statistics were analyzed using the Spearman correlation. (e) The proliferative responses were evaluated by detection of CFSE lo cells in IgM - lymphocytes by flow cytometry ( n = 12). Representative dot plots of CFSE-staining in lymphocytes other than B cells are shown. Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P

    Article Snippet: After being blocked with 3% skim milk in phosphate-buffered saline (PBS, pH 7.2) containing 0.05% Tween 20 (PBS-T), the membranes were incubated at room temperature for 2 h with anti-PD-1 mAb (2C12 and 3G2: 3 μg/mL, 2H7 and 5D2: 1 μg/mL), followed by washing and incubation with horse radish peroxidase (HRP)-conjugated anti-rat IgG (MP Biomedicals, CA, USA).

    Techniques: Cell Function Assay, Cell Culture, Negative Control, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Cytometry, Staining

    Recognition of PD-1-expressing lymphocytes by anti-PD-1 mAb. (a) Gating strategies for CD4 + T cells, CD8 + T cells and IgM + B cells in bovine lymphocytes. (b) Representative histograms obtained by flow cytometry of PD-1 expression in CD4 + T cells, CD8 + T cells and IgM + B cells isolated from three BLV - cattle. Freshly isolated PBMC were stained with anti-PD-1 (5D2), CD4, CD8, and IgM Ab. PBMC were cultivated with PBS (No stimulation), PMA/ionomycin, and PWM for 24, 48, and 72 h, and stained in a similar way. (c) Proportions of PD-1 positive cells in CD4 + T cells, CD8 + T cells and IgM + B cells. Statistical comparisons between percentages of PD-1 positive cells stimulated with PBS and PMA/ionomycin or PWM were made using two way ANOVA. Differences were considered statistically significant at P

    Journal: Veterinary Research

    Article Title: Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vitro

    doi: 10.1186/1297-9716-44-59

    Figure Lengend Snippet: Recognition of PD-1-expressing lymphocytes by anti-PD-1 mAb. (a) Gating strategies for CD4 + T cells, CD8 + T cells and IgM + B cells in bovine lymphocytes. (b) Representative histograms obtained by flow cytometry of PD-1 expression in CD4 + T cells, CD8 + T cells and IgM + B cells isolated from three BLV - cattle. Freshly isolated PBMC were stained with anti-PD-1 (5D2), CD4, CD8, and IgM Ab. PBMC were cultivated with PBS (No stimulation), PMA/ionomycin, and PWM for 24, 48, and 72 h, and stained in a similar way. (c) Proportions of PD-1 positive cells in CD4 + T cells, CD8 + T cells and IgM + B cells. Statistical comparisons between percentages of PD-1 positive cells stimulated with PBS and PMA/ionomycin or PWM were made using two way ANOVA. Differences were considered statistically significant at P

    Article Snippet: After being blocked with 3% skim milk in phosphate-buffered saline (PBS, pH 7.2) containing 0.05% Tween 20 (PBS-T), the membranes were incubated at room temperature for 2 h with anti-PD-1 mAb (2C12 and 3G2: 3 μg/mL, 2H7 and 5D2: 1 μg/mL), followed by washing and incubation with horse radish peroxidase (HRP)-conjugated anti-rat IgG (MP Biomedicals, CA, USA).

    Techniques: Expressing, Flow Cytometry, Cytometry, Isolation, Staining

    Enhancement of cytokine production by anti-PD-1 mAb in bovine lymphocytes. PBMC were cultivated with rat IgG control or four types of anti-PD-1 mAb (20 μg/mL) in the absence ( a ; n = 8 or 14) or presence ( b ; n = 16) of PMA/ionomycin. IFN-γ production was measured by ELISA. Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P

    Journal: Veterinary Research

    Article Title: Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vitro

    doi: 10.1186/1297-9716-44-59

    Figure Lengend Snippet: Enhancement of cytokine production by anti-PD-1 mAb in bovine lymphocytes. PBMC were cultivated with rat IgG control or four types of anti-PD-1 mAb (20 μg/mL) in the absence ( a ; n = 8 or 14) or presence ( b ; n = 16) of PMA/ionomycin. IFN-γ production was measured by ELISA. Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P

    Article Snippet: After being blocked with 3% skim milk in phosphate-buffered saline (PBS, pH 7.2) containing 0.05% Tween 20 (PBS-T), the membranes were incubated at room temperature for 2 h with anti-PD-1 mAb (2C12 and 3G2: 3 μg/mL, 2H7 and 5D2: 1 μg/mL), followed by washing and incubation with horse radish peroxidase (HRP)-conjugated anti-rat IgG (MP Biomedicals, CA, USA).

    Techniques: Enzyme-linked Immunosorbent Assay

    Expression analysis of bovine PD-1 in BLV-infected cattle. (a, b) Percentages of PD-1-expressing CD4 + and CD8 + T cells in PBMC (a) and LN (b) isolated from BLV - cattle (CD4 + T cells; n = 20 and 31, CD8 + T cells; n = 16 and 31), BLV + cattle (CD4 + T cells; n = 35 and 15, CD8 + T cells; n = 28 and 15) and BCBL (CD4 + T cells; n = 7 and 6, CD8 + T cells; n = 7 and 6). (c) An example of gating strategies for PD-1 expression in CD4 + T cells and CD8 + T cells isolated from lymph node of BCBL and BLV - cattle. Values in the quadrant indicate the percentage of the cells in lymphocytes other than IgM + B cells. (d) Percentages of PD-L1-expressing gp51 + ( n = 5) and gp51 - ( n = 5) in IgM + B cells from BLV + and BLV - cattle ( n = 9). Representative dot plots of PD-L1 staining in gp51 + or gp51 - B cells are also shown. Statistical comparisons were made using one-way ANOVA with Tukey’s test. Differences were considered statistically significant at P

    Journal: Veterinary Research

    Article Title: Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vitro

    doi: 10.1186/1297-9716-44-59

    Figure Lengend Snippet: Expression analysis of bovine PD-1 in BLV-infected cattle. (a, b) Percentages of PD-1-expressing CD4 + and CD8 + T cells in PBMC (a) and LN (b) isolated from BLV - cattle (CD4 + T cells; n = 20 and 31, CD8 + T cells; n = 16 and 31), BLV + cattle (CD4 + T cells; n = 35 and 15, CD8 + T cells; n = 28 and 15) and BCBL (CD4 + T cells; n = 7 and 6, CD8 + T cells; n = 7 and 6). (c) An example of gating strategies for PD-1 expression in CD4 + T cells and CD8 + T cells isolated from lymph node of BCBL and BLV - cattle. Values in the quadrant indicate the percentage of the cells in lymphocytes other than IgM + B cells. (d) Percentages of PD-L1-expressing gp51 + ( n = 5) and gp51 - ( n = 5) in IgM + B cells from BLV + and BLV - cattle ( n = 9). Representative dot plots of PD-L1 staining in gp51 + or gp51 - B cells are also shown. Statistical comparisons were made using one-way ANOVA with Tukey’s test. Differences were considered statistically significant at P

    Article Snippet: After being blocked with 3% skim milk in phosphate-buffered saline (PBS, pH 7.2) containing 0.05% Tween 20 (PBS-T), the membranes were incubated at room temperature for 2 h with anti-PD-1 mAb (2C12 and 3G2: 3 μg/mL, 2H7 and 5D2: 1 μg/mL), followed by washing and incubation with horse radish peroxidase (HRP)-conjugated anti-rat IgG (MP Biomedicals, CA, USA).

    Techniques: Expressing, Infection, Isolation, Staining

    Effect of PD-1 blockade on gp51 expression and B-cell activation. (a , b, c) Percentages of gp51-expressing cells ( a ; n = 15) and mean fluorescence index (MFI) of WC4 (CD19 like molecule) ( b ; n = 12) and CD80 ( c ; n = 6) in IgM + B cells were evaluated by flow cytometry in PBMC treated with rat IgG control or anti-PD-1 mAb (20 μg/mL). Representative contour plots showing gp51 expression (right panels) in PBMC treated with rat IgG control (upper panels) or anti-PD-1 mAb (lower panels) is shown in (a) . No staining was observed in PBMC stained with isotype control for anti-gp51 mAb (left panels). (d) Expression of BAFF mRNA was evaluated by real-time PCR ( n = 12). The results are indicated as relative change to control (no antibody treatment) when the amount of BAFF mRNA expression is divided by GAPDH mRNA expression. (e) Percentages of apoptotic cells in IgM + B cells were measured by flow cytometry. Apoptotic B cells were identified as annexin-V + 7-AAD - cells ( n = 11). (f) Percentages of gp51-expressing cells were evaluated by flow cytometry in isolated B cells cultivated with rat IgG control or anti-PD-1 mAb ( n = 11). Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P

    Journal: Veterinary Research

    Article Title: Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vitro

    doi: 10.1186/1297-9716-44-59

    Figure Lengend Snippet: Effect of PD-1 blockade on gp51 expression and B-cell activation. (a , b, c) Percentages of gp51-expressing cells ( a ; n = 15) and mean fluorescence index (MFI) of WC4 (CD19 like molecule) ( b ; n = 12) and CD80 ( c ; n = 6) in IgM + B cells were evaluated by flow cytometry in PBMC treated with rat IgG control or anti-PD-1 mAb (20 μg/mL). Representative contour plots showing gp51 expression (right panels) in PBMC treated with rat IgG control (upper panels) or anti-PD-1 mAb (lower panels) is shown in (a) . No staining was observed in PBMC stained with isotype control for anti-gp51 mAb (left panels). (d) Expression of BAFF mRNA was evaluated by real-time PCR ( n = 12). The results are indicated as relative change to control (no antibody treatment) when the amount of BAFF mRNA expression is divided by GAPDH mRNA expression. (e) Percentages of apoptotic cells in IgM + B cells were measured by flow cytometry. Apoptotic B cells were identified as annexin-V + 7-AAD - cells ( n = 11). (f) Percentages of gp51-expressing cells were evaluated by flow cytometry in isolated B cells cultivated with rat IgG control or anti-PD-1 mAb ( n = 11). Statistical comparisons between rat IgG control and anti-PD-1 mAb were made using the Wilcoxon matched-pairs test. Differences were considered statistically significant at P

    Article Snippet: After being blocked with 3% skim milk in phosphate-buffered saline (PBS, pH 7.2) containing 0.05% Tween 20 (PBS-T), the membranes were incubated at room temperature for 2 h with anti-PD-1 mAb (2C12 and 3G2: 3 μg/mL, 2H7 and 5D2: 1 μg/mL), followed by washing and incubation with horse radish peroxidase (HRP)-conjugated anti-rat IgG (MP Biomedicals, CA, USA).

    Techniques: Expressing, Activation Assay, Fluorescence, Flow Cytometry, Cytometry, Staining, Real-time Polymerase Chain Reaction, Isolation

    Recognition of PD-1-expressing cells by anti-PD-1 mAb. (a) Flow cytometric analysis of surface expression of bovine PD-1. Cos-7 expressing PD-1 (black line) and Cos-7 transfected with the control vector (dashed line) were stained with four types of anti-PD-1 mAb (2C12, 2H7, 3G2, and 5D2) and isotype control (Rat IgG). (b) Western blotting analysis of bovine PD-1 expression in CHO-DG44 cells stably expressing bovine PD-1. Three types of anti-PD-1 mAb recognized PD-1 (triangle) at about 68 kDa. Anti-actin antibody and anti-myc antibody was used as a loading control and a positive control.

    Journal: Veterinary Research

    Article Title: Blockade of bovine PD-1 increases T cell function and inhibits bovine leukemia virus expression in B cells in vitro

    doi: 10.1186/1297-9716-44-59

    Figure Lengend Snippet: Recognition of PD-1-expressing cells by anti-PD-1 mAb. (a) Flow cytometric analysis of surface expression of bovine PD-1. Cos-7 expressing PD-1 (black line) and Cos-7 transfected with the control vector (dashed line) were stained with four types of anti-PD-1 mAb (2C12, 2H7, 3G2, and 5D2) and isotype control (Rat IgG). (b) Western blotting analysis of bovine PD-1 expression in CHO-DG44 cells stably expressing bovine PD-1. Three types of anti-PD-1 mAb recognized PD-1 (triangle) at about 68 kDa. Anti-actin antibody and anti-myc antibody was used as a loading control and a positive control.

    Article Snippet: After being blocked with 3% skim milk in phosphate-buffered saline (PBS, pH 7.2) containing 0.05% Tween 20 (PBS-T), the membranes were incubated at room temperature for 2 h with anti-PD-1 mAb (2C12 and 3G2: 3 μg/mL, 2H7 and 5D2: 1 μg/mL), followed by washing and incubation with horse radish peroxidase (HRP)-conjugated anti-rat IgG (MP Biomedicals, CA, USA).

    Techniques: Expressing, Flow Cytometry, Transfection, Plasmid Preparation, Staining, Western Blot, Stable Transfection, Positive Control