monoclonal anti-cd40 Search Results


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  • N/A
    This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified recombinant Human CD40 extracellular domain
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    97
    Bio X Cell anti cd40
    Host conditioning with <t>anti-CD40</t> induces high-level T cell accumulation in the lymphoid organs and brains of SV11 mice at early time points. Groups of mice received either anti-CD40, WBI, or no conditioning regimen with TCR-IV T-cell ACT. Representative plots show MHC tetramer staining (mean±SEM) of TCR-IV T cells on days +4 and +5 in a spleen and c brain. Quantification of TCR-IV T-cell accumulation (mean±SEM) in b spleen and d brain of mice that received ACT with the indicated treatments. n=3 mice/group (except n=2 for day +4 control group). Data shown are from one experiment and representative of two independent experiments. Asterisks above connecting lines indicate significant differences between time points. Asterisks next to vertical brackets indicate significant differences between treatment groups. * p
    Anti Cd40, supplied by Bio X Cell, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd40/product/Bio X Cell
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cd40 - by Bioz Stars, 2021-04
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    95
    BioLegend anti cd40
    Effect of ICOSL on CD40L presentation and reception in SLB model for T FH cell- GC B cell interaction. a , Activated human T cells that express ICOS and CD40L were incubated with SLB containing ICAM-1 and UCHT1 (anti-CD3) as a basal condition with a ring of ICAM-1 surrounding a central cluster enriched in T cell receptor enriched extracellular vesicles by 15 minutes 26 . This condition resulted in low presentation of CD40L in punctate structures detected by anti-CD40L mAb that accumulated in the same central synapse with the TCR enriched extracellular vesicles. Addition of ICOSL the SLB resulted in strong central accumulation of fluorescent ICOSL with the TCR enriched extracellular vesicles, but no increase in CD40L presentation. Addition of <t>CD40</t> the SLB resulted in a significant increase in CD40L accumulation, which we refer to as reception because its receptor dependent. When ICOSL and CD40 were added the reception of CD40L was further significantly enhanced over the level observed with CD40 alone. Thus, ICOSL ligation in the centre of the immunological synapse increases CD40L reception. All levels are shown in gray scale except CD40L panels, for which the pseudocolor scale is indicated. Scale bar 5 µm. b , Human T FH cells were incubated with SLB containing ICAM-1 and UCHT1 (anti-CD3). Addition of ICOSL resulted in increased accumulation of CgB at the synapse centre. Addition of CD40 did not further increased CgB accumulation.
    Anti Cd40, supplied by BioLegend, 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/anti cd40/product/BioLegend
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cd40 - by Bioz Stars, 2021-04
    95/100 stars
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    93
    BioLegend cd40
    Effect of ICOSL on CD40L presentation and reception in SLB model for T FH cell- GC B cell interaction. a , Activated human T cells that express ICOS and CD40L were incubated with SLB containing ICAM-1 and UCHT1 (anti-CD3) as a basal condition with a ring of ICAM-1 surrounding a central cluster enriched in T cell receptor enriched extracellular vesicles by 15 minutes 26 . This condition resulted in low presentation of CD40L in punctate structures detected by anti-CD40L mAb that accumulated in the same central synapse with the TCR enriched extracellular vesicles. Addition of ICOSL the SLB resulted in strong central accumulation of fluorescent ICOSL with the TCR enriched extracellular vesicles, but no increase in CD40L presentation. Addition of <t>CD40</t> the SLB resulted in a significant increase in CD40L accumulation, which we refer to as reception because its receptor dependent. When ICOSL and CD40 were added the reception of CD40L was further significantly enhanced over the level observed with CD40 alone. Thus, ICOSL ligation in the centre of the immunological synapse increases CD40L reception. All levels are shown in gray scale except CD40L panels, for which the pseudocolor scale is indicated. Scale bar 5 µm. b , Human T FH cells were incubated with SLB containing ICAM-1 and UCHT1 (anti-CD3). Addition of ICOSL resulted in increased accumulation of CgB at the synapse centre. Addition of CD40 did not further increased CgB accumulation.
    Cd40, supplied by BioLegend, 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/cd40/product/BioLegend
    Average 93 stars, based on 1 article reviews
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    cd40 - by Bioz Stars, 2021-04
    93/100 stars
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    97
    Cell Signaling Technology Inc anti human antibodies
    Effect of ICOSL on CD40L presentation and reception in SLB model for T FH cell- GC B cell interaction. a , Activated human T cells that express ICOS and CD40L were incubated with SLB containing ICAM-1 and UCHT1 (anti-CD3) as a basal condition with a ring of ICAM-1 surrounding a central cluster enriched in T cell receptor enriched extracellular vesicles by 15 minutes 26 . This condition resulted in low presentation of CD40L in punctate structures detected by anti-CD40L mAb that accumulated in the same central synapse with the TCR enriched extracellular vesicles. Addition of ICOSL the SLB resulted in strong central accumulation of fluorescent ICOSL with the TCR enriched extracellular vesicles, but no increase in CD40L presentation. Addition of <t>CD40</t> the SLB resulted in a significant increase in CD40L accumulation, which we refer to as reception because its receptor dependent. When ICOSL and CD40 were added the reception of CD40L was further significantly enhanced over the level observed with CD40 alone. Thus, ICOSL ligation in the centre of the immunological synapse increases CD40L reception. All levels are shown in gray scale except CD40L panels, for which the pseudocolor scale is indicated. Scale bar 5 µm. b , Human T FH cells were incubated with SLB containing ICAM-1 and UCHT1 (anti-CD3). Addition of ICOSL resulted in increased accumulation of CgB at the synapse centre. Addition of CD40 did not further increased CgB accumulation.
    Anti Human Antibodies, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti human antibodies/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti human antibodies - by Bioz Stars, 2021-04
    97/100 stars
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    N/A
    This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified recombinant Human CD40 TNFRSF5 rh
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    Image Search Results


    Host conditioning with anti-CD40 induces high-level T cell accumulation in the lymphoid organs and brains of SV11 mice at early time points. Groups of mice received either anti-CD40, WBI, or no conditioning regimen with TCR-IV T-cell ACT. Representative plots show MHC tetramer staining (mean±SEM) of TCR-IV T cells on days +4 and +5 in a spleen and c brain. Quantification of TCR-IV T-cell accumulation (mean±SEM) in b spleen and d brain of mice that received ACT with the indicated treatments. n=3 mice/group (except n=2 for day +4 control group). Data shown are from one experiment and representative of two independent experiments. Asterisks above connecting lines indicate significant differences between time points. Asterisks next to vertical brackets indicate significant differences between treatment groups. * p

    Journal: Cancer immunology, immunotherapy : CII

    Article Title: Protection from tumor recurrence following adoptive immunotherapy varies with host conditioning regimen despite initial regression of autochthonous murine brain tumors

    doi: 10.1007/s00262-014-1635-7

    Figure Lengend Snippet: Host conditioning with anti-CD40 induces high-level T cell accumulation in the lymphoid organs and brains of SV11 mice at early time points. Groups of mice received either anti-CD40, WBI, or no conditioning regimen with TCR-IV T-cell ACT. Representative plots show MHC tetramer staining (mean±SEM) of TCR-IV T cells on days +4 and +5 in a spleen and c brain. Quantification of TCR-IV T-cell accumulation (mean±SEM) in b spleen and d brain of mice that received ACT with the indicated treatments. n=3 mice/group (except n=2 for day +4 control group). Data shown are from one experiment and representative of two independent experiments. Asterisks above connecting lines indicate significant differences between time points. Asterisks next to vertical brackets indicate significant differences between treatment groups. * p

    Article Snippet: Anti-CD40- and control-conditioned mice received 100 μg purified anti-CD40 (clone FGK45, BioXcell) or control rat IgG (Sigma) on days −1 and +1 by intraperitoneal (i.p.) injection.

    Techniques: Mouse Assay, Activated Clotting Time Assay, Staining

    Anti-CD40-enhanced ACT promotes initial regression of established tumors. a H E brain sections on day +10 post-ACT following conditioning with anti-CD40 (left), control IgG (middle), or WBI (right). Representative low-power images (top row, scale bar = 1mm) and high-power images (bottom row, scale bar = 50μm) that show established tumor refractory to therapy (middle column) or tumor stromal condensation indicative of tumor regression (left and right columns). b For each mouse, the largest cross-sectional tumor area (mm 2 ) observed in H E sections was plotted. Data is pooled from multiple experiments with a total of 6–10 mice/group. Statistical significance was determined using the Kruskal-Wallis test with Dunn’s multiple comparison test. ** p

    Journal: Cancer immunology, immunotherapy : CII

    Article Title: Protection from tumor recurrence following adoptive immunotherapy varies with host conditioning regimen despite initial regression of autochthonous murine brain tumors

    doi: 10.1007/s00262-014-1635-7

    Figure Lengend Snippet: Anti-CD40-enhanced ACT promotes initial regression of established tumors. a H E brain sections on day +10 post-ACT following conditioning with anti-CD40 (left), control IgG (middle), or WBI (right). Representative low-power images (top row, scale bar = 1mm) and high-power images (bottom row, scale bar = 50μm) that show established tumor refractory to therapy (middle column) or tumor stromal condensation indicative of tumor regression (left and right columns). b For each mouse, the largest cross-sectional tumor area (mm 2 ) observed in H E sections was plotted. Data is pooled from multiple experiments with a total of 6–10 mice/group. Statistical significance was determined using the Kruskal-Wallis test with Dunn’s multiple comparison test. ** p

    Article Snippet: Anti-CD40- and control-conditioned mice received 100 μg purified anti-CD40 (clone FGK45, BioXcell) or control rat IgG (Sigma) on days −1 and +1 by intraperitoneal (i.p.) injection.

    Techniques: Activated Clotting Time Assay, Mouse Assay

    Anti-CD40-enhanced ACT promotes increased survival but short-term surveillance against tumor recurrence. a Groups of mice received the indicated conditioning with or without ACT and were monitored for tumor recurrence. The percentage of surviving mice versus age is plotted. b Statistical differences in survival were calculated using the log-rank test. Data are pooled from multiple experiments with 5–8 mice/group.

    Journal: Cancer immunology, immunotherapy : CII

    Article Title: Protection from tumor recurrence following adoptive immunotherapy varies with host conditioning regimen despite initial regression of autochthonous murine brain tumors

    doi: 10.1007/s00262-014-1635-7

    Figure Lengend Snippet: Anti-CD40-enhanced ACT promotes increased survival but short-term surveillance against tumor recurrence. a Groups of mice received the indicated conditioning with or without ACT and were monitored for tumor recurrence. The percentage of surviving mice versus age is plotted. b Statistical differences in survival were calculated using the log-rank test. Data are pooled from multiple experiments with 5–8 mice/group.

    Article Snippet: Anti-CD40- and control-conditioned mice received 100 μg purified anti-CD40 (clone FGK45, BioXcell) or control rat IgG (Sigma) on days −1 and +1 by intraperitoneal (i.p.) injection.

    Techniques: Activated Clotting Time Assay, Mouse Assay

    Donor T cells fail to persist in anti-CD40-conditioned SV11 mice following acute tumor regression. Groups of SV11 mice received either anti-CD40, control IgG, or WBI conditioning prior to ACT with CD90.1 + TCR-IV T cells. On day +30, cells from a spleens, cLN (not shown), and b brains were stained for CD90.1 and CD8. Values on dot plots indicate percent CD90.1 + of total CD8 + cells (mean±SEM). c Total CD90.1 + cells in spleens, cLN, and brains on day +30 are plotted. Representative histograms of CD44, CD62L, and KLRG1 expression gated on live CD45.2 + CD8 + CD90.1 + TCR-IV T cells (open histogram) or CD45.2 + CD8 + CD90.1 − T cells (filled histogram, spleen only) are shown on day +30 in d spleen and e brain. Values indicate the percent of TCR-IV T cells within the indicated gate (mean±SEM). Samples with

    Journal: Cancer immunology, immunotherapy : CII

    Article Title: Protection from tumor recurrence following adoptive immunotherapy varies with host conditioning regimen despite initial regression of autochthonous murine brain tumors

    doi: 10.1007/s00262-014-1635-7

    Figure Lengend Snippet: Donor T cells fail to persist in anti-CD40-conditioned SV11 mice following acute tumor regression. Groups of SV11 mice received either anti-CD40, control IgG, or WBI conditioning prior to ACT with CD90.1 + TCR-IV T cells. On day +30, cells from a spleens, cLN (not shown), and b brains were stained for CD90.1 and CD8. Values on dot plots indicate percent CD90.1 + of total CD8 + cells (mean±SEM). c Total CD90.1 + cells in spleens, cLN, and brains on day +30 are plotted. Representative histograms of CD44, CD62L, and KLRG1 expression gated on live CD45.2 + CD8 + CD90.1 + TCR-IV T cells (open histogram) or CD45.2 + CD8 + CD90.1 − T cells (filled histogram, spleen only) are shown on day +30 in d spleen and e brain. Values indicate the percent of TCR-IV T cells within the indicated gate (mean±SEM). Samples with

    Article Snippet: Anti-CD40- and control-conditioned mice received 100 μg purified anti-CD40 (clone FGK45, BioXcell) or control rat IgG (Sigma) on days −1 and +1 by intraperitoneal (i.p.) injection.

    Techniques: Mouse Assay, Activated Clotting Time Assay, Staining, Expressing

    Effect of ICOSL on CD40L presentation and reception in SLB model for T FH cell- GC B cell interaction. a , Activated human T cells that express ICOS and CD40L were incubated with SLB containing ICAM-1 and UCHT1 (anti-CD3) as a basal condition with a ring of ICAM-1 surrounding a central cluster enriched in T cell receptor enriched extracellular vesicles by 15 minutes 26 . This condition resulted in low presentation of CD40L in punctate structures detected by anti-CD40L mAb that accumulated in the same central synapse with the TCR enriched extracellular vesicles. Addition of ICOSL the SLB resulted in strong central accumulation of fluorescent ICOSL with the TCR enriched extracellular vesicles, but no increase in CD40L presentation. Addition of CD40 the SLB resulted in a significant increase in CD40L accumulation, which we refer to as reception because its receptor dependent. When ICOSL and CD40 were added the reception of CD40L was further significantly enhanced over the level observed with CD40 alone. Thus, ICOSL ligation in the centre of the immunological synapse increases CD40L reception. All levels are shown in gray scale except CD40L panels, for which the pseudocolor scale is indicated. Scale bar 5 µm. b , Human T FH cells were incubated with SLB containing ICAM-1 and UCHT1 (anti-CD3). Addition of ICOSL resulted in increased accumulation of CgB at the synapse centre. Addition of CD40 did not further increased CgB accumulation.

    Journal: Nature

    Article Title: TFH-derived dopamine accelerates productive synapses in germinal centres

    doi: 10.1038/nature23013

    Figure Lengend Snippet: Effect of ICOSL on CD40L presentation and reception in SLB model for T FH cell- GC B cell interaction. a , Activated human T cells that express ICOS and CD40L were incubated with SLB containing ICAM-1 and UCHT1 (anti-CD3) as a basal condition with a ring of ICAM-1 surrounding a central cluster enriched in T cell receptor enriched extracellular vesicles by 15 minutes 26 . This condition resulted in low presentation of CD40L in punctate structures detected by anti-CD40L mAb that accumulated in the same central synapse with the TCR enriched extracellular vesicles. Addition of ICOSL the SLB resulted in strong central accumulation of fluorescent ICOSL with the TCR enriched extracellular vesicles, but no increase in CD40L presentation. Addition of CD40 the SLB resulted in a significant increase in CD40L accumulation, which we refer to as reception because its receptor dependent. When ICOSL and CD40 were added the reception of CD40L was further significantly enhanced over the level observed with CD40 alone. Thus, ICOSL ligation in the centre of the immunological synapse increases CD40L reception. All levels are shown in gray scale except CD40L panels, for which the pseudocolor scale is indicated. Scale bar 5 µm. b , Human T FH cells were incubated with SLB containing ICAM-1 and UCHT1 (anti-CD3). Addition of ICOSL resulted in increased accumulation of CgB at the synapse centre. Addition of CD40 did not further increased CgB accumulation.

    Article Snippet: For the experiment with dopamine receptor block, 2 x 105 sorted germinal centre B cells were stimulated with 5 μM of freshly prepared DA with or without 50nM of Haloperidol (Tocris) for 2 h. Incubation media was then replaced with fresh media containing anti-CD40 (1 μg/ml, BioLegend) and IL-21 (10 ng/ml, Peprotech), cells were incubated for 5 days and plasma cell differentiation was assessed.

    Techniques: Incubation, Ligation

    Dopamine is released from T FH cells upon cognate interactions. a-c Flow cytometric quantification of dopamine content in FSK-stimulated T FH cells after 30 min incubation with anti-CD3/CD28 beads (1:1) or autologous or allogeneic GC B cells (1:2) (n=3) ( b ) also showing changes in DA content in GC B cells (autologous or allogeneic) cultured separately (“nil”), or together with T FH cells (n=5); and with or without ICAM-1 (5 μg/ml) and LFA-1 (10 μg/ml) block ( c ) (n=3); Mann-Whitney test. d , Flow cytometric plots showing plasma cells (PCs), identified as CD27 hi CD38 hi , induced in cultures of GC B cells stimulated for five days with anti-CD40 (2 μg/ml), IL-21 (20 ng/ml) and different concentrations of freshly-prepared DA (n=5). e , Fold changes in PC differentiation from GC B cells stimulated for 2h with or without DA (5μM) and Haloperidol (Haldol, 50nM), and cultured in the presence of anti-CD40 (2 μg/ml) and IL-21 (20 ng/ml) for 5d; two tailed student t-test. a-c, e: Bars represent median values and each dot represents a single experiment conducted in triplicates (n=5). Two tailed student t-test; ns, not significant, *p ≤ 0.05, ***p ≤ 0.001.

    Journal: Nature

    Article Title: TFH-derived dopamine accelerates productive synapses in germinal centres

    doi: 10.1038/nature23013

    Figure Lengend Snippet: Dopamine is released from T FH cells upon cognate interactions. a-c Flow cytometric quantification of dopamine content in FSK-stimulated T FH cells after 30 min incubation with anti-CD3/CD28 beads (1:1) or autologous or allogeneic GC B cells (1:2) (n=3) ( b ) also showing changes in DA content in GC B cells (autologous or allogeneic) cultured separately (“nil”), or together with T FH cells (n=5); and with or without ICAM-1 (5 μg/ml) and LFA-1 (10 μg/ml) block ( c ) (n=3); Mann-Whitney test. d , Flow cytometric plots showing plasma cells (PCs), identified as CD27 hi CD38 hi , induced in cultures of GC B cells stimulated for five days with anti-CD40 (2 μg/ml), IL-21 (20 ng/ml) and different concentrations of freshly-prepared DA (n=5). e , Fold changes in PC differentiation from GC B cells stimulated for 2h with or without DA (5μM) and Haloperidol (Haldol, 50nM), and cultured in the presence of anti-CD40 (2 μg/ml) and IL-21 (20 ng/ml) for 5d; two tailed student t-test. a-c, e: Bars represent median values and each dot represents a single experiment conducted in triplicates (n=5). Two tailed student t-test; ns, not significant, *p ≤ 0.05, ***p ≤ 0.001.

    Article Snippet: For the experiment with dopamine receptor block, 2 x 105 sorted germinal centre B cells were stimulated with 5 μM of freshly prepared DA with or without 50nM of Haloperidol (Tocris) for 2 h. Incubation media was then replaced with fresh media containing anti-CD40 (1 μg/ml, BioLegend) and IL-21 (10 ng/ml, Peprotech), cells were incubated for 5 days and plasma cell differentiation was assessed.

    Techniques: Flow Cytometry, Incubation, Cell Culture, Blocking Assay, MANN-WHITNEY, Two Tailed Test

    Dopamine induces ICOSL upregulation on human GC B cells. a , Gating of GC B cells and fluorescence intensity of specified proteins 30 minutes after stimulation with DA (10μM) (n=3). b , Fold changes of surface ICOSL expression with medium control set as unit 1 (n=8). c , Survival of GC B cells after DA stimulation (n=8). d , Fold changes of surface ICOSL expression on GC B cells stimulated with DA (10μM), DA agonist SKF38393 (10nM), Haloperidol (50nM) and DA antagonist SKF83566 (10nM) for 30 min, with medium control set as unit 1 (n=5). e , f , Representative histograms ( e ) and quantification ( f ) of surface and intracellular ICOSL on naïve, memory and GC B cells (n=4); Mann-Whitney test. g , RNA counts per million (CPM) of indicated transcripts in human GC B cells stimulated with or without DA (5μM) for 2h (n=3). h , Fold changes of surface ICOSL expression on human GC B cells treated with cycloheximide (CHX, 10 μg/ml) and stimulated with DA (10μM) for 30 min. i, j, Fold changes of surface ICOSL expression on human GC B cells stimulated with DA (10μM), anti-CD40 (1 μg/ml) or recombinant CD40L (10 μg/ml) ( i ), IL-21 (10, 50 or 100 ng/ml) or IL-4 (10 νγ/ml) ( j ) for 30 min (n=5). b, d, h-j , Bars represent medians and each dot represents a single experiment conducted in triplicate (n=10); two tailed student t-test. ns, not significant, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 and ****p ≤ 0.0001.

    Journal: Nature

    Article Title: TFH-derived dopamine accelerates productive synapses in germinal centres

    doi: 10.1038/nature23013

    Figure Lengend Snippet: Dopamine induces ICOSL upregulation on human GC B cells. a , Gating of GC B cells and fluorescence intensity of specified proteins 30 minutes after stimulation with DA (10μM) (n=3). b , Fold changes of surface ICOSL expression with medium control set as unit 1 (n=8). c , Survival of GC B cells after DA stimulation (n=8). d , Fold changes of surface ICOSL expression on GC B cells stimulated with DA (10μM), DA agonist SKF38393 (10nM), Haloperidol (50nM) and DA antagonist SKF83566 (10nM) for 30 min, with medium control set as unit 1 (n=5). e , f , Representative histograms ( e ) and quantification ( f ) of surface and intracellular ICOSL on naïve, memory and GC B cells (n=4); Mann-Whitney test. g , RNA counts per million (CPM) of indicated transcripts in human GC B cells stimulated with or without DA (5μM) for 2h (n=3). h , Fold changes of surface ICOSL expression on human GC B cells treated with cycloheximide (CHX, 10 μg/ml) and stimulated with DA (10μM) for 30 min. i, j, Fold changes of surface ICOSL expression on human GC B cells stimulated with DA (10μM), anti-CD40 (1 μg/ml) or recombinant CD40L (10 μg/ml) ( i ), IL-21 (10, 50 or 100 ng/ml) or IL-4 (10 νγ/ml) ( j ) for 30 min (n=5). b, d, h-j , Bars represent medians and each dot represents a single experiment conducted in triplicate (n=10); two tailed student t-test. ns, not significant, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 and ****p ≤ 0.0001.

    Article Snippet: For the experiment with dopamine receptor block, 2 x 105 sorted germinal centre B cells were stimulated with 5 μM of freshly prepared DA with or without 50nM of Haloperidol (Tocris) for 2 h. Incubation media was then replaced with fresh media containing anti-CD40 (1 μg/ml, BioLegend) and IL-21 (10 ng/ml, Peprotech), cells were incubated for 5 days and plasma cell differentiation was assessed.

    Techniques: Fluorescence, Expressing, MANN-WHITNEY, Recombinant, Two Tailed Test

    Regulation of ICOSL upregulation in mouse and human B cells a, Fold changes of surface ICOSL expression on mouse GC B cells that were treated with anti-CD40 (10 μg/ml) and DA (0.5, 1, 5, 10μM) for 30 minutes, with medium control set as unit 1 (n=5). b, Representative histogram and quantification of surface and intracellular ICOSL on GC and non-GC B cells (n=5). **p ≤ 0.01; nonparametric Mann-Whitney test (U test). c, RNA counts per million of ICOSL, CD40, BCL6, IL21R, CD86, BAFFR and FAS mRNA in human memory B cells stimulated with or without DA (5μM) for 2h (n=3). d , Fold changes of surface ICOSL expression on mouse GC B cells that were treated with cycloheximide (CHX, 10 μg/ml) for 4h, with medium control set as unit 1. Bars represent median values and each dot represents a single mouse. e, Fold changes of surface ICOSL expression on mouse GC B cells that were stimulated with BAFF (100ng/ml), LPS (1 or 10 μg/ml), anti-CD40 (10 μg/ml) and anti-IgM (1 or 10 μg/ml) for 30 min and 4h. Unit 1 set on medium control. f, Fold changes of surface ICOSL expression on mouse GC B cells that were treated with actinomycin D (ActD, 5 μg/ml), anti-CD40 (10 μg/ml) for 4h, with medium control set as unit 1. Bars represent median and each dot represent a single mouse (n=5). d-f , ns, not significant, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 and ****p ≤ 0.0001; two tailed student t-test. g , Representative histogram of surface ICOSL expression on human GC B cells that were stimulated with DA (10μM) or anti-CD40 (1 μg/ml) for 30 min. h , Fold changes of surface ICOSL expression on human GC B cells stimulated with several concentrations of anti-CD40 for 4 and 8 hours, with medium control set as unit 1 (n=3). i , Bar plot showing survival of GC B in the presence of anti-CD40 (1 μg/ml) after 4 or 8 hours of stimulation (n=4). *p ≤ 0.05 and ***p ≤ 0.001; nonparametric Mann-Whitney test (U test).

    Journal: Nature

    Article Title: TFH-derived dopamine accelerates productive synapses in germinal centres

    doi: 10.1038/nature23013

    Figure Lengend Snippet: Regulation of ICOSL upregulation in mouse and human B cells a, Fold changes of surface ICOSL expression on mouse GC B cells that were treated with anti-CD40 (10 μg/ml) and DA (0.5, 1, 5, 10μM) for 30 minutes, with medium control set as unit 1 (n=5). b, Representative histogram and quantification of surface and intracellular ICOSL on GC and non-GC B cells (n=5). **p ≤ 0.01; nonparametric Mann-Whitney test (U test). c, RNA counts per million of ICOSL, CD40, BCL6, IL21R, CD86, BAFFR and FAS mRNA in human memory B cells stimulated with or without DA (5μM) for 2h (n=3). d , Fold changes of surface ICOSL expression on mouse GC B cells that were treated with cycloheximide (CHX, 10 μg/ml) for 4h, with medium control set as unit 1. Bars represent median values and each dot represents a single mouse. e, Fold changes of surface ICOSL expression on mouse GC B cells that were stimulated with BAFF (100ng/ml), LPS (1 or 10 μg/ml), anti-CD40 (10 μg/ml) and anti-IgM (1 or 10 μg/ml) for 30 min and 4h. Unit 1 set on medium control. f, Fold changes of surface ICOSL expression on mouse GC B cells that were treated with actinomycin D (ActD, 5 μg/ml), anti-CD40 (10 μg/ml) for 4h, with medium control set as unit 1. Bars represent median and each dot represent a single mouse (n=5). d-f , ns, not significant, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 and ****p ≤ 0.0001; two tailed student t-test. g , Representative histogram of surface ICOSL expression on human GC B cells that were stimulated with DA (10μM) or anti-CD40 (1 μg/ml) for 30 min. h , Fold changes of surface ICOSL expression on human GC B cells stimulated with several concentrations of anti-CD40 for 4 and 8 hours, with medium control set as unit 1 (n=3). i , Bar plot showing survival of GC B in the presence of anti-CD40 (1 μg/ml) after 4 or 8 hours of stimulation (n=4). *p ≤ 0.05 and ***p ≤ 0.001; nonparametric Mann-Whitney test (U test).

    Article Snippet: For the experiment with dopamine receptor block, 2 x 105 sorted germinal centre B cells were stimulated with 5 μM of freshly prepared DA with or without 50nM of Haloperidol (Tocris) for 2 h. Incubation media was then replaced with fresh media containing anti-CD40 (1 μg/ml, BioLegend) and IL-21 (10 ng/ml, Peprotech), cells were incubated for 5 days and plasma cell differentiation was assessed.

    Techniques: Expressing, MANN-WHITNEY, Mann-Whitney U-Test, Two Tailed Test

    Effects of ICOS ligation at the immunological synapse. a , Representative images of ICAM-1 ring (white) around CD40L (pseudocolor scale) in the presence or absence of CD40 and ICOSL at physiological densities on the supported lipid bilayer (SLB) containing ICAM-1 and UCHT1. Scale bar 5 µm. b , c , Plots represent CD40L MFI of individual activated human T ( b ) or T FH ( c ) cells forming synapses (n=3). d , Representative images of chromogranin B stain in the presence or absence of ICOSL at the immunological synapse. e , Plots represent CgB fluorescent intensity of individual activated T FH and non-T FH cells forming synapses (n=3). b,c,e , ns, not significant, (***p ≤ 0.001) and (****p ≤ 0.0001) nonparametric Mann-Whitney test (U test). f , Representative images of CgB + T FH cells (red) forming synapses with allogeneic B cells (green).

    Journal: Nature

    Article Title: TFH-derived dopamine accelerates productive synapses in germinal centres

    doi: 10.1038/nature23013

    Figure Lengend Snippet: Effects of ICOS ligation at the immunological synapse. a , Representative images of ICAM-1 ring (white) around CD40L (pseudocolor scale) in the presence or absence of CD40 and ICOSL at physiological densities on the supported lipid bilayer (SLB) containing ICAM-1 and UCHT1. Scale bar 5 µm. b , c , Plots represent CD40L MFI of individual activated human T ( b ) or T FH ( c ) cells forming synapses (n=3). d , Representative images of chromogranin B stain in the presence or absence of ICOSL at the immunological synapse. e , Plots represent CgB fluorescent intensity of individual activated T FH and non-T FH cells forming synapses (n=3). b,c,e , ns, not significant, (***p ≤ 0.001) and (****p ≤ 0.0001) nonparametric Mann-Whitney test (U test). f , Representative images of CgB + T FH cells (red) forming synapses with allogeneic B cells (green).

    Article Snippet: For the experiment with dopamine receptor block, 2 x 105 sorted germinal centre B cells were stimulated with 5 μM of freshly prepared DA with or without 50nM of Haloperidol (Tocris) for 2 h. Incubation media was then replaced with fresh media containing anti-CD40 (1 μg/ml, BioLegend) and IL-21 (10 ng/ml, Peprotech), cells were incubated for 5 days and plasma cell differentiation was assessed.

    Techniques: Ligation, Staining, MANN-WHITNEY, Mann-Whitney U-Test