Review

cd40l blocking mab (Enzo Biochem)

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Enzo Biochem cd40l blocking mab

(A) Bright field images (400×) of mature DC1 or DC2 co-cultured in the presence of SEB alone (left panels) or with CD4+ T cells (middle panels), or rhCD40L (right panels) for 24 h. The arrows highlight TNT-like extensions. (B) 24 h co-cultures of SEB Ag-presenting DC1 and CD4+ T cells in the presence of control mAb (top) or <t>CD40L</t> blocking mAb (bottom). (C) Representative Z-series projection image revealing a network of TNT-like membrane connections in fluorescently labeled DC1. (A-C) Images are representative of 6 independent experiments conducted on DC from 3 healthy donors. (D) Representative Z-series projection images of a CD40L-activated DC1 analyzed using the IMARIS ‘Surfaces’ program to determine total cell surface area (left panel), and IMARIS ‘FilamentTracer’ to trace the pathway of each ‘filament’ branch extending from a single origin (blue sphere) at the edge of the cell body (middle panel). Individual segments begin at an origin or branch point (orange spheres) and end at the next branch or terminal point (green spheres). Spatial ‘reach’ was defined as the shortest distance from a filament origin to the distal terminal point of that filament (right panel; white lines connecting 2 points). (E) Total cell surface area comparisons of resting and CD40L-activated DC1 and DC2. (F) Comparison of percentage ‘TNT positive’ resting DC1 and DC2 (media only), defined as those expressing ≥5 individual TNTs per cell that were each >5.0 μm in length. (G) Comparison of percentage ‘reticulation positive’ CD40L-activated DC1 and DC2, delineated as those displaying ≥5 filaments per cell that were each >10.0 μm in sum segment length. (H) ‘Maximum reach’ of filaments, defined as the shortest distance (μm) from the origin to the farthest terminal point of a filament. (E-H) Data were generated from randomly chosen image fields (20–30 per donor) and represented as mean ± SD of 3 healthy donors independently tested. P-values <0.0001, <0.001, <0.01 and <0.05 are represented by ****, ***, **, and *, respectively. See also Supplemental Fig. 1; Video 1; Video 2.

Cd40l Blocking Mab, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cd40l+blocking+mab/pmc04297732-102-2-5?v=Enzo+Biochem
Average 90 stars, based on 1 article reviews

cd40l blocking mab - by Bioz Stars, 2026-07

90/100 stars

Images

1) Product Images from "CD40L induces functional tunneling nanotube networks exclusively in dendritic cells programmed by mediators of type-1 immunity"

Article Title: CD40L induces functional tunneling nanotube networks exclusively in dendritic cells programmed by mediators of type-1 immunity

Journal: Journal of immunology (Baltimore, Md. : 1950)

doi: 10.4049/jimmunol.1401832

Figure Legend Snippet: (A) Bright field images (400×) of mature DC1 or DC2 co-cultured in the presence of SEB alone (left panels) or with CD4+ T cells (middle panels), or rhCD40L (right panels) for 24 h. The arrows highlight TNT-like extensions. (B) 24 h co-cultures of SEB Ag-presenting DC1 and CD4+ T cells in the presence of control mAb (top) or CD40L blocking mAb (bottom). (C) Representative Z-series projection image revealing a network of TNT-like membrane connections in fluorescently labeled DC1. (A-C) Images are representative of 6 independent experiments conducted on DC from 3 healthy donors. (D) Representative Z-series projection images of a CD40L-activated DC1 analyzed using the IMARIS ‘Surfaces’ program to determine total cell surface area (left panel), and IMARIS ‘FilamentTracer’ to trace the pathway of each ‘filament’ branch extending from a single origin (blue sphere) at the edge of the cell body (middle panel). Individual segments begin at an origin or branch point (orange spheres) and end at the next branch or terminal point (green spheres). Spatial ‘reach’ was defined as the shortest distance from a filament origin to the distal terminal point of that filament (right panel; white lines connecting 2 points). (E) Total cell surface area comparisons of resting and CD40L-activated DC1 and DC2. (F) Comparison of percentage ‘TNT positive’ resting DC1 and DC2 (media only), defined as those expressing ≥5 individual TNTs per cell that were each >5.0 μm in length. (G) Comparison of percentage ‘reticulation positive’ CD40L-activated DC1 and DC2, delineated as those displaying ≥5 filaments per cell that were each >10.0 μm in sum segment length. (H) ‘Maximum reach’ of filaments, defined as the shortest distance (μm) from the origin to the farthest terminal point of a filament. (E-H) Data were generated from randomly chosen image fields (20–30 per donor) and represented as mean ± SD of 3 healthy donors independently tested. P-values <0.0001, <0.001, <0.01 and <0.05 are represented by ****, ***, **, and *, respectively. See also Supplemental Fig. 1; Video 1; Video 2.

Techniques Used: Cell Culture, Blocking Assay, Labeling, Expressing, Generated

(A–C) 3D reconstruction images (600×) of differentially matured DC stimulated for 20 h with rhCD40L prior to labeling the cell surface with MHC class I mAb (green) and labeling nuclei (blue), followed by live-cell confocal imaging. Data are representative of 6 independent experiments conducted using DC from 3 healthy donors. (A) Membrane morphologies of mature CD40L-activated DC0, DC1, and DC2 propagated using LPS alone, LPS + IFN-γ, or LPS + PGE2, respectively. (B) Mature CD40L-treated DC0, DC1, and DC2 generated by the respective use of R848 alone, R848 + IFN-γ, or R848 + PGE2. (C) Cell morphologies of DC1 treated with media alone or CD40L. DC1 were generated using the αDC1 cytokine-based cocktail (first two panels on left), or induced by 48 h co-culture of iDC with either 2-signal activated NKh cells (middle) or SEB-activated CD8+ T cells (right).

Figure Legend Snippet: (A–C) 3D reconstruction images (600×) of differentially matured DC stimulated for 20 h with rhCD40L prior to labeling the cell surface with MHC class I mAb (green) and labeling nuclei (blue), followed by live-cell confocal imaging. Data are representative of 6 independent experiments conducted using DC from 3 healthy donors. (A) Membrane morphologies of mature CD40L-activated DC0, DC1, and DC2 propagated using LPS alone, LPS + IFN-γ, or LPS + PGE2, respectively. (B) Mature CD40L-treated DC0, DC1, and DC2 generated by the respective use of R848 alone, R848 + IFN-γ, or R848 + PGE2. (C) Cell morphologies of DC1 treated with media alone or CD40L. DC1 were generated using the αDC1 cytokine-based cocktail (first two panels on left), or induced by 48 h co-culture of iDC with either 2-signal activated NKh cells (middle) or SEB-activated CD8+ T cells (right).

Techniques Used: Labeling, Imaging, Generated, Co-Culture Assay

(A) Sequential still frames of DC1 actively reticulating from 4 to 7.6 h post-addition of rhCD40L, captured by live-cell, time-lapse DIC imaging (600×). See also Video 3. (B) Sequential frames of high resolution, time-lapse DIC imaging (600×) of live 8 h rhCD40L-stimulated DC1 showing endogenous cell structures resembling vesicles (arrows) trafficking between neighboring cells through CD40L-induced TNTs. See also Video 4. (C) Confocal reconstruction images (1000×) revealing early endosome- (green, arrows) and F-actin-containing TNTs (red) and nuclei (blue) in fixed CD40L-activated DC1. Data are representative of 3 donors independently tested.

Figure Legend Snippet: (A) Sequential still frames of DC1 actively reticulating from 4 to 7.6 h post-addition of rhCD40L, captured by live-cell, time-lapse DIC imaging (600×). See also Video 3. (B) Sequential frames of high resolution, time-lapse DIC imaging (600×) of live 8 h rhCD40L-stimulated DC1 showing endogenous cell structures resembling vesicles (arrows) trafficking between neighboring cells through CD40L-induced TNTs. See also Video 4. (C) Confocal reconstruction images (1000×) revealing early endosome- (green, arrows) and F-actin-containing TNTs (red) and nuclei (blue) in fixed CD40L-activated DC1. Data are representative of 3 donors independently tested.

Techniques Used: Imaging

(A–C) Cy5-labeled (blue), YG latex bead (40 nm)-containing ‘donor’ DC1 or DC2 were co-cultured with the respective non-bead-containing, Cy3-labeled (red) ‘recipient’ DC type for 20 h in the presence or absence of rhCD40L. Data are representative of 2 donors independently tested. (A) Montage of time-lapse, confocal imaging (600×) showing beads (green; arrows) moving rapidly through donor DC1 TNTs (blue) over a time span of 28 min. See also Video 5. (B) Montage tracing the pathway of beads (green; arrows) traveling from the cell body of donor DC1 (blue), through donor cell TNTs and into the connected recipient DC (red), where they finally collect in the recipient cell body (yellow, lower left arrows). (C) Flow cytometric analysis and quantification of intercellular exogenous bead transfer from donor to recipient DC types after treatment with media only or CD40L. (D) IFNγ-ELISPOT assays measuring Ag-specific recall responses of cultured T cells following their in vitro sensitization with FACS sorted, YG bead/Ag positive and negative recipient DC1. Data are represented as mean ± SE of 2 independent experiments, and * represents the p-value <0.05. See also Supplemental Fig. 3.

Figure Legend Snippet: (A–C) Cy5-labeled (blue), YG latex bead (40 nm)-containing ‘donor’ DC1 or DC2 were co-cultured with the respective non-bead-containing, Cy3-labeled (red) ‘recipient’ DC type for 20 h in the presence or absence of rhCD40L. Data are representative of 2 donors independently tested. (A) Montage of time-lapse, confocal imaging (600×) showing beads (green; arrows) moving rapidly through donor DC1 TNTs (blue) over a time span of 28 min. See also Video 5. (B) Montage tracing the pathway of beads (green; arrows) traveling from the cell body of donor DC1 (blue), through donor cell TNTs and into the connected recipient DC (red), where they finally collect in the recipient cell body (yellow, lower left arrows). (C) Flow cytometric analysis and quantification of intercellular exogenous bead transfer from donor to recipient DC types after treatment with media only or CD40L. (D) IFNγ-ELISPOT assays measuring Ag-specific recall responses of cultured T cells following their in vitro sensitization with FACS sorted, YG bead/Ag positive and negative recipient DC1. Data are represented as mean ± SE of 2 independent experiments, and * represents the p-value <0.05. See also Supplemental Fig. 3.

Techniques Used: Labeling, Cell Culture, Imaging, Enzyme-linked Immunospot, In Vitro

(A) DC1 monolayers that had been stimulated with rhCD40L for 10 h were exposed to EGFP-expressing Escherichia coli for 2 h. Subsequent time-lapse confocal resonant scanning microscopy shows individual bacterium (green) trafficking bi-directionally along CD40L-induced TNTs (arrows) between mature DC1. See also Video 6. (B) Donor DC1 (blue) pulsed with EGFP-expressing HIV-1-like particles (green) were co-cultured with recipient DC1 (red) in the presence of CD40L for 10 h prior to live-cell imaging. HIV-1-like particles localizing to a donor cell nanotube (blue; arrows) that has formed a connection with a recipient cell body. (C) At 20 h post-CD40L stimulation, HIV-1-like particles can be detected at the interface between blue donor cell TNTs and red recipient cells (arrows), prior to their subsequent transfer to recipient cell bodies. (A-C) Imaging data are representative of 3 donors independently tested.

Figure Legend Snippet: (A) DC1 monolayers that had been stimulated with rhCD40L for 10 h were exposed to EGFP-expressing Escherichia coli for 2 h. Subsequent time-lapse confocal resonant scanning microscopy shows individual bacterium (green) trafficking bi-directionally along CD40L-induced TNTs (arrows) between mature DC1. See also Video 6. (B) Donor DC1 (blue) pulsed with EGFP-expressing HIV-1-like particles (green) were co-cultured with recipient DC1 (red) in the presence of CD40L for 10 h prior to live-cell imaging. HIV-1-like particles localizing to a donor cell nanotube (blue; arrows) that has formed a connection with a recipient cell body. (C) At 20 h post-CD40L stimulation, HIV-1-like particles can be detected at the interface between blue donor cell TNTs and red recipient cells (arrows), prior to their subsequent transfer to recipient cell bodies. (A-C) Imaging data are representative of 3 donors independently tested.

Techniques Used: Expressing, Microscopy, Cell Culture, Live Cell Imaging, Imaging

Image Search Results

a Comparison of the total number of intrahepatic effector gBT-1 T cells at 14 days post-rAAV treatment in recipient mice treated with anti-CD4 depleting, anti-CD40L blocking Ab, or isotype control Ab. b Comparison of the total number of intrahepatic effector gBT-1 T cells at 15 days post-rAAV treatment of recipient mice treated with isotype control, or CD40L blocking Abs beginning day 0 or day 9 post-rAAV. c Comparison of the total number of intrahepatic effector gBT-1 T cells at 15 days post-rAAV treatment of CD4-depleted recipient mice administered agonist anti-CD40 mAb or isotype control 9-days post-rAAV. d Representative IF image of a portal tract cluster containing transferred gBT-1 and P25 T cells interacting with MHCII high cells in the liver 12-days post-rAAV treatment of recipient mice. Data are representative of two independent experiments. n = 4 ( a ) or n = 5 ( b ), n = 4 or 5 ( c ), n = 3 mice per group. Error bars indicate ± SEM. Analysed with one-way ANOVA with Tukey’s multiple comparisons test ( a , b ), or a two-tailed Student’s t test (c). Scale bars, 40 μm/20 μm ( d ). Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: A hepatic network of dendritic cells mediates CD4 T cell help outside lymphoid organs

doi: 10.1038/s41467-024-45612-5

Figure Lengend Snippet: a Comparison of the total number of intrahepatic effector gBT-1 T cells at 14 days post-rAAV treatment in recipient mice treated with anti-CD4 depleting, anti-CD40L blocking Ab, or isotype control Ab. b Comparison of the total number of intrahepatic effector gBT-1 T cells at 15 days post-rAAV treatment of recipient mice treated with isotype control, or CD40L blocking Abs beginning day 0 or day 9 post-rAAV. c Comparison of the total number of intrahepatic effector gBT-1 T cells at 15 days post-rAAV treatment of CD4-depleted recipient mice administered agonist anti-CD40 mAb or isotype control 9-days post-rAAV. d Representative IF image of a portal tract cluster containing transferred gBT-1 and P25 T cells interacting with MHCII high cells in the liver 12-days post-rAAV treatment of recipient mice. Data are representative of two independent experiments. n = 4 ( a ) or n = 5 ( b ), n = 4 or 5 ( c ), n = 3 mice per group. Error bars indicate ± SEM. Analysed with one-way ANOVA with Tukey’s multiple comparisons test ( a , b ), or a two-tailed Student’s t test (c). Scale bars, 40 μm/20 μm ( d ). Source data are provided as a Source Data file.

Article Snippet: To block CD40L in vivo, mice were injected i.v. with 500 μg MR1 anti-mouse CD40L blocking mAb (BioXcell, West Lebanon, USA) or polyclonal Armenian hamster IgG isotype control (BioXcell, West Lebanon, USA) in PBS at the indicated time points.

Techniques: Comparison, Blocking Assay, Control, Two Tailed Test

a – c Representative confocal IF images of the liver of XCR1 Venus/+ rAAV-treated recipient mice at 12-days post rAAV, showing large clusters of XCR1 + cDC1s, gBT-1 and P25 T cells in PT regions Scale bars 50 μm ( a ), 30 μm ( b ), 4 μm/2 μm ( c ). d , e Quantification of the average distance (μm) separating XCR1 + cells with gBT-1 T cells and ( d ), or with P25 T cells ( e ) in portal tracts (PT), peri-central vein (PCV) and sinusoidal (S) liver regions. Data points represent a single region of interest from n = 7 mice from 2 independent experiments. f , g Hepatic XCR1 + cDC1 isolated from recipient mice treated, 12-days prior, with rAAV gB-Ag85b or PBS, were assessed for their ability to activate and induce proliferation (assessed by CD44 upregulation and CTV dilution), of naive gBT-1 ( f ) or P25 ( g ) T cells co-cultured for 72 h. h , i Same as ( f , g ) but recipient mice were also treated with isotype control (black) or anti-CD40L blocking mAb (red) to assess the role of CD40/CD40L interactions. Representative flow plots ( h ), and proportion of proliferating CD44 high CTV low gBT-1 T cells ( i ). j Co-stimulatory molecule expression levels in hepatic cDC1s isolated from rAAV gB-Ag85b –treated mice that also received isotype control (black) or anti-CD4 depleting mAb (red) at 12-days post rAAV. k , l Quantification of Ki67 + gB T-1 T cell numbers forming clusters in PT ( k ) and PCV regions ( l ) at 12-days post-rAAV treatment in XCR1 +/+ (black) and XCR1 DTR/+ (red) recipient mice treated with DTx, 9-days prior. m , n Quantification of total numbers of intrahepatic ( m ) and splenic ( n ) effector gBT-1 cell in recipient mice treated with rAAV, 15-days prior. Comparison between diphtheria toxin treated XCR1 +/+ (black) and XCR1 DTR/+ (red) recipient mice at 9-days post-rAAV.Data representative of two independent experiments. n = 5 ( a – c ), n = 7 ( d , e ), n = 3 or 4 ( f , g ), n = 7 ( i ), n = 4 ( j ) n = 5 ( k , l ) and n = 6 ( m , n ) mice per group. Error bars indicate ± SEM. Analysed with a Kruskal-Wallis test with multiple comparisons ( d , e ), a two-tailed unpaired Student’s t test ( i , j ) or a two tailed unpaired Mann–Whitney U test ( k – n ); ns not statistically significant. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: A hepatic network of dendritic cells mediates CD4 T cell help outside lymphoid organs

doi: 10.1038/s41467-024-45612-5

Figure Lengend Snippet: a – c Representative confocal IF images of the liver of XCR1 Venus/+ rAAV-treated recipient mice at 12-days post rAAV, showing large clusters of XCR1 + cDC1s, gBT-1 and P25 T cells in PT regions Scale bars 50 μm ( a ), 30 μm ( b ), 4 μm/2 μm ( c ). d , e Quantification of the average distance (μm) separating XCR1 + cells with gBT-1 T cells and ( d ), or with P25 T cells ( e ) in portal tracts (PT), peri-central vein (PCV) and sinusoidal (S) liver regions. Data points represent a single region of interest from n = 7 mice from 2 independent experiments. f , g Hepatic XCR1 + cDC1 isolated from recipient mice treated, 12-days prior, with rAAV gB-Ag85b or PBS, were assessed for their ability to activate and induce proliferation (assessed by CD44 upregulation and CTV dilution), of naive gBT-1 ( f ) or P25 ( g ) T cells co-cultured for 72 h. h , i Same as ( f , g ) but recipient mice were also treated with isotype control (black) or anti-CD40L blocking mAb (red) to assess the role of CD40/CD40L interactions. Representative flow plots ( h ), and proportion of proliferating CD44 high CTV low gBT-1 T cells ( i ). j Co-stimulatory molecule expression levels in hepatic cDC1s isolated from rAAV gB-Ag85b –treated mice that also received isotype control (black) or anti-CD4 depleting mAb (red) at 12-days post rAAV. k , l Quantification of Ki67 + gB T-1 T cell numbers forming clusters in PT ( k ) and PCV regions ( l ) at 12-days post-rAAV treatment in XCR1 +/+ (black) and XCR1 DTR/+ (red) recipient mice treated with DTx, 9-days prior. m , n Quantification of total numbers of intrahepatic ( m ) and splenic ( n ) effector gBT-1 cell in recipient mice treated with rAAV, 15-days prior. Comparison between diphtheria toxin treated XCR1 +/+ (black) and XCR1 DTR/+ (red) recipient mice at 9-days post-rAAV.Data representative of two independent experiments. n = 5 ( a – c ), n = 7 ( d , e ), n = 3 or 4 ( f , g ), n = 7 ( i ), n = 4 ( j ) n = 5 ( k , l ) and n = 6 ( m , n ) mice per group. Error bars indicate ± SEM. Analysed with a Kruskal-Wallis test with multiple comparisons ( d , e ), a two-tailed unpaired Student’s t test ( i , j ) or a two tailed unpaired Mann–Whitney U test ( k – n ); ns not statistically significant. Source data are provided as a Source Data file.

Techniques: Isolation, Cell Culture, Control, Blocking Assay, Expressing, Comparison, Two Tailed Test, MANN-WHITNEY

Differential expression of Satb1 in TCR75 T cells from tolerant and rejecting mice. ( A ) Experimental design. Congenic TCR75 CD4 + T cells seeded 1 d before transplantation were sorted on day 35 posttransplantation from spleen and lymph nodes (LNs) or allografts of tolerant or rejecting mice and RNA-seq was performed. ( B ) Principal component analysis. ( C ) Heatmap of differentially expressed transcription factors (fold change ≥ 1.5; adjusted P ≤ 0.05). Color key is normalized log2 transformed counts per million (CPM). n = 3 (memory: TCR75 T cells from 1 mouse/replicate; tolerant: pooled from 5 mice/replicate, except for tolerant allograft n = 1 with TCR75 T cells pooled from 15 allografts). ( D and E ) TCR75 T cells were seeded into host B6 mice 1 d before transplantation with B/c heart ±donor splenocytes and anti-CD154. Mice were euthanized at induction of memory and tolerance (day 3 ± 1 and day 12 ± 2) and Satb1 protein expression (MFI) was measured by flow cytometry of TCR75-gated T cells in spleen ( D ) and allograft ( E ). Naive group in ( D and E ) is the same and shows Satb1 expression of splenic TCR75 T cells before transfer. ( F ) Congenic OTII T cells were seeded 1 d before transplantation of 2W-OVA-expressing skin grafts, and hosts were treated or not with anti-CD154 and 2W-OVA-expressing splenocytes. OTII cells were sorted from spleen on d35 posttransplantation and analyzed by flow cytometry for expression of Satb1. All results are displayed as mean +/− SEM. n = 3 to 9/group ( D ), n = 3 to 5/group ( E ) and n = 3 to 4/group ( F ). * P < 0.05; ** P < 0.01 (ANOVA, two-way for D , one-way for E and F , with pairwise comparisons).

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Reduced Satb1 expression predisposes CD4 + T conventional cells to Treg suppression and promotes transplant survival

doi: 10.1073/pnas.2205062119

Figure Lengend Snippet: Differential expression of Satb1 in TCR75 T cells from tolerant and rejecting mice. ( A ) Experimental design. Congenic TCR75 CD4 + T cells seeded 1 d before transplantation were sorted on day 35 posttransplantation from spleen and lymph nodes (LNs) or allografts of tolerant or rejecting mice and RNA-seq was performed. ( B ) Principal component analysis. ( C ) Heatmap of differentially expressed transcription factors (fold change ≥ 1.5; adjusted P ≤ 0.05). Color key is normalized log2 transformed counts per million (CPM). n = 3 (memory: TCR75 T cells from 1 mouse/replicate; tolerant: pooled from 5 mice/replicate, except for tolerant allograft n = 1 with TCR75 T cells pooled from 15 allografts). ( D and E ) TCR75 T cells were seeded into host B6 mice 1 d before transplantation with B/c heart ±donor splenocytes and anti-CD154. Mice were euthanized at induction of memory and tolerance (day 3 ± 1 and day 12 ± 2) and Satb1 protein expression (MFI) was measured by flow cytometry of TCR75-gated T cells in spleen ( D ) and allograft ( E ). Naive group in ( D and E ) is the same and shows Satb1 expression of splenic TCR75 T cells before transfer. ( F ) Congenic OTII T cells were seeded 1 d before transplantation of 2W-OVA-expressing skin grafts, and hosts were treated or not with anti-CD154 and 2W-OVA-expressing splenocytes. OTII cells were sorted from spleen on d35 posttransplantation and analyzed by flow cytometry for expression of Satb1. All results are displayed as mean +/− SEM. n = 3 to 9/group ( D ), n = 3 to 5/group ( E ) and n = 3 to 4/group ( F ). * P < 0.05; ** P < 0.01 (ANOVA, two-way for D , one-way for E and F , with pairwise comparisons).

Article Snippet: For induction of tolerance, heart recipients received an intravenous injection of splenocytes from 1/4 of donor spleen on day 0 and were treated with 600 μg of anti-CD154 (MR1) blocking mAb (Bio-X-cell) on days 0 (intravenous), 7 (intraperitoneal), and 14 (intraperitoneal) postheart transplantation.

Techniques: Quantitative Proteomics, Transplantation Assay, RNA Sequencing, Transformation Assay, Expressing, Flow Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD40L induces functional tunneling nanotube networks exclusively in dendritic cells programmed by mediators of type-1 immunity

doi: 10.4049/jimmunol.1401832

Figure Lengend Snippet: (A) Bright field images (400×) of mature DC1 or DC2 co-cultured in the presence of SEB alone (left panels) or with CD4+ T cells (middle panels), or rhCD40L (right panels) for 24 h. The arrows highlight TNT-like extensions. (B) 24 h co-cultures of SEB Ag-presenting DC1 and CD4+ T cells in the presence of control mAb (top) or CD40L blocking mAb (bottom). (C) Representative Z-series projection image revealing a network of TNT-like membrane connections in fluorescently labeled DC1. (A-C) Images are representative of 6 independent experiments conducted on DC from 3 healthy donors. (D) Representative Z-series projection images of a CD40L-activated DC1 analyzed using the IMARIS ‘Surfaces’ program to determine total cell surface area (left panel), and IMARIS ‘FilamentTracer’ to trace the pathway of each ‘filament’ branch extending from a single origin (blue sphere) at the edge of the cell body (middle panel). Individual segments begin at an origin or branch point (orange spheres) and end at the next branch or terminal point (green spheres). Spatial ‘reach’ was defined as the shortest distance from a filament origin to the distal terminal point of that filament (right panel; white lines connecting 2 points). (E) Total cell surface area comparisons of resting and CD40L-activated DC1 and DC2. (F) Comparison of percentage ‘TNT positive’ resting DC1 and DC2 (media only), defined as those expressing ≥5 individual TNTs per cell that were each >5.0 μm in length. (G) Comparison of percentage ‘reticulation positive’ CD40L-activated DC1 and DC2, delineated as those displaying ≥5 filaments per cell that were each >10.0 μm in sum segment length. (H) ‘Maximum reach’ of filaments, defined as the shortest distance (μm) from the origin to the farthest terminal point of a filament. (E-H) Data were generated from randomly chosen image fields (20–30 per donor) and represented as mean ± SD of 3 healthy donors independently tested. P-values <0.0001, <0.001, <0.01 and <0.05 are represented by ****, ***, **, and *, respectively. See also Supplemental Fig. 1; Video 1; Video 2.

Article Snippet: When used, CD40L blocking mAb (Enzo Life Sciences) or control mAb (BD Biosciences, San Jose, CA) were added to the cultures.

Techniques: Cell Culture, Blocking Assay, Labeling, Expressing, Generated

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD40L induces functional tunneling nanotube networks exclusively in dendritic cells programmed by mediators of type-1 immunity

doi: 10.4049/jimmunol.1401832

Figure Lengend Snippet: (A–C) 3D reconstruction images (600×) of differentially matured DC stimulated for 20 h with rhCD40L prior to labeling the cell surface with MHC class I mAb (green) and labeling nuclei (blue), followed by live-cell confocal imaging. Data are representative of 6 independent experiments conducted using DC from 3 healthy donors. (A) Membrane morphologies of mature CD40L-activated DC0, DC1, and DC2 propagated using LPS alone, LPS + IFN-γ, or LPS + PGE2, respectively. (B) Mature CD40L-treated DC0, DC1, and DC2 generated by the respective use of R848 alone, R848 + IFN-γ, or R848 + PGE2. (C) Cell morphologies of DC1 treated with media alone or CD40L. DC1 were generated using the αDC1 cytokine-based cocktail (first two panels on left), or induced by 48 h co-culture of iDC with either 2-signal activated NKh cells (middle) or SEB-activated CD8+ T cells (right).

Article Snippet: When used, CD40L blocking mAb (Enzo Life Sciences) or control mAb (BD Biosciences, San Jose, CA) were added to the cultures.

Techniques: Labeling, Imaging, Generated, Co-Culture Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD40L induces functional tunneling nanotube networks exclusively in dendritic cells programmed by mediators of type-1 immunity

doi: 10.4049/jimmunol.1401832

Figure Lengend Snippet: (A) Sequential still frames of DC1 actively reticulating from 4 to 7.6 h post-addition of rhCD40L, captured by live-cell, time-lapse DIC imaging (600×). See also Video 3. (B) Sequential frames of high resolution, time-lapse DIC imaging (600×) of live 8 h rhCD40L-stimulated DC1 showing endogenous cell structures resembling vesicles (arrows) trafficking between neighboring cells through CD40L-induced TNTs. See also Video 4. (C) Confocal reconstruction images (1000×) revealing early endosome- (green, arrows) and F-actin-containing TNTs (red) and nuclei (blue) in fixed CD40L-activated DC1. Data are representative of 3 donors independently tested.

Article Snippet: When used, CD40L blocking mAb (Enzo Life Sciences) or control mAb (BD Biosciences, San Jose, CA) were added to the cultures.

Techniques: Imaging

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD40L induces functional tunneling nanotube networks exclusively in dendritic cells programmed by mediators of type-1 immunity

doi: 10.4049/jimmunol.1401832

Figure Lengend Snippet: (A–C) Cy5-labeled (blue), YG latex bead (40 nm)-containing ‘donor’ DC1 or DC2 were co-cultured with the respective non-bead-containing, Cy3-labeled (red) ‘recipient’ DC type for 20 h in the presence or absence of rhCD40L. Data are representative of 2 donors independently tested. (A) Montage of time-lapse, confocal imaging (600×) showing beads (green; arrows) moving rapidly through donor DC1 TNTs (blue) over a time span of 28 min. See also Video 5. (B) Montage tracing the pathway of beads (green; arrows) traveling from the cell body of donor DC1 (blue), through donor cell TNTs and into the connected recipient DC (red), where they finally collect in the recipient cell body (yellow, lower left arrows). (C) Flow cytometric analysis and quantification of intercellular exogenous bead transfer from donor to recipient DC types after treatment with media only or CD40L. (D) IFNγ-ELISPOT assays measuring Ag-specific recall responses of cultured T cells following their in vitro sensitization with FACS sorted, YG bead/Ag positive and negative recipient DC1. Data are represented as mean ± SE of 2 independent experiments, and * represents the p-value <0.05. See also Supplemental Fig. 3.

Article Snippet: When used, CD40L blocking mAb (Enzo Life Sciences) or control mAb (BD Biosciences, San Jose, CA) were added to the cultures.

Techniques: Labeling, Cell Culture, Imaging, Enzyme-linked Immunospot, In Vitro

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD40L induces functional tunneling nanotube networks exclusively in dendritic cells programmed by mediators of type-1 immunity

doi: 10.4049/jimmunol.1401832

Figure Lengend Snippet: (A) DC1 monolayers that had been stimulated with rhCD40L for 10 h were exposed to EGFP-expressing Escherichia coli for 2 h. Subsequent time-lapse confocal resonant scanning microscopy shows individual bacterium (green) trafficking bi-directionally along CD40L-induced TNTs (arrows) between mature DC1. See also Video 6. (B) Donor DC1 (blue) pulsed with EGFP-expressing HIV-1-like particles (green) were co-cultured with recipient DC1 (red) in the presence of CD40L for 10 h prior to live-cell imaging. HIV-1-like particles localizing to a donor cell nanotube (blue; arrows) that has formed a connection with a recipient cell body. (C) At 20 h post-CD40L stimulation, HIV-1-like particles can be detected at the interface between blue donor cell TNTs and red recipient cells (arrows), prior to their subsequent transfer to recipient cell bodies. (A-C) Imaging data are representative of 3 donors independently tested.

Article Snippet: When used, CD40L blocking mAb (Enzo Life Sciences) or control mAb (BD Biosciences, San Jose, CA) were added to the cultures.

Techniques: Expressing, Microscopy, Cell Culture, Live Cell Imaging, Imaging

Anti-CD40L MAb 5c8 inhibits IFN-γ production in cocultures of PBL and M. tuberculosis-infected MN. In order to confirm the ability of 5c8 to adequately block CD40L-CD40 interactions in our culture system, we measured IFN-γ production by cocultured M. tuberculosis-infected MN and PBL both in medium alone and in the presence of several concentrations of anti-CD40L. As indicated, the addition of antiCD40L led to a dose-dependent decrease in measured IFN-γ. Mean IFN-γ levels were significantly lower than control levels in the presence of 1, 10, and 100 μg of 5c8/ml (P = 0.001, 0.021, and 0.007, respectively). Bars represent means and standard deviations of results from studies of 10 subjects.

Journal:

Article Title: CD40 Ligand (CD154) Does Not Contribute to Lymphocyte-Mediated Inhibition of Virulent Mycobacterium tuberculosis within Human Monocytes

doi: 10.1128/IAI.70.8.4716-4720.2002

Figure Lengend Snippet: Anti-CD40L MAb 5c8 inhibits IFN-γ production in cocultures of PBL and M. tuberculosis-infected MN. In order to confirm the ability of 5c8 to adequately block CD40L-CD40 interactions in our culture system, we measured IFN-γ production by cocultured M. tuberculosis-infected MN and PBL both in medium alone and in the presence of several concentrations of anti-CD40L. As indicated, the addition of antiCD40L led to a dose-dependent decrease in measured IFN-γ. Mean IFN-γ levels were significantly lower than control levels in the presence of 1, 10, and 100 μg of 5c8/ml (P = 0.001, 0.021, and 0.007, respectively). Bars represent means and standard deviations of results from studies of 10 subjects.

Article Snippet: To investigate this possibility, we studied the ability of blocking anti-CD40L monoclonal antibody (MAb) 5c8 (kindly provided by Biogen, Inc.) to inhibit lymphocyte-mediated limitation of the growth of H37Rv within MN.

Techniques: Infection, Blocking Assay, Control

Blocking anti-CD40L MAb 5c8 has no effect on the ability of PBL to limit the intracellular growth of M. tuberculosis H37Rv. The figure illustrates CFU of H37Rv within MN immediately following infection and at 1, 4, and 7 days. Infected MN were cocultured with PBL alone and with PBL plus 5c8 in concentrations of 1, 10, and 100 μg/ml. Anti-CD40L did not significantly alter the ability of PBL to limit the intracellular growth of M. tuberculosis at any of the time points. Results illustrate means and standard deviations of results for cells from 10 donors.

Journal:

Article Title: CD40 Ligand (CD154) Does Not Contribute to Lymphocyte-Mediated Inhibition of Virulent Mycobacterium tuberculosis within Human Monocytes

doi: 10.1128/IAI.70.8.4716-4720.2002

Figure Lengend Snippet: Blocking anti-CD40L MAb 5c8 has no effect on the ability of PBL to limit the intracellular growth of M. tuberculosis H37Rv. The figure illustrates CFU of H37Rv within MN immediately following infection and at 1, 4, and 7 days. Infected MN were cocultured with PBL alone and with PBL plus 5c8 in concentrations of 1, 10, and 100 μg/ml. Anti-CD40L did not significantly alter the ability of PBL to limit the intracellular growth of M. tuberculosis at any of the time points. Results illustrate means and standard deviations of results for cells from 10 donors.

Techniques: Blocking Assay, Infection

Anti-CD40L MAb does not inhibit killing of intracellular M. tuberculosis by antigen-specific CD4+ T cells. CD4+ T cells were isolated from PBMC that had been stimulated for 7 days by incubation with avirulent M. tuberculosis H37Ra. As illustrated, the addition of these cells in a 5:1 ratio relative to infected MN resulted in a significant reduction of intracellular M. tuberculosis at 1, 4, and 7 days of coculture. The addition of 5c8 in concentrations of 10 and 100 μg/ml had no effect on CD4+ T-cell-mediated killing of H37Rv.

Journal:

Article Title: CD40 Ligand (CD154) Does Not Contribute to Lymphocyte-Mediated Inhibition of Virulent Mycobacterium tuberculosis within Human Monocytes

doi: 10.1128/IAI.70.8.4716-4720.2002

Figure Lengend Snippet: Anti-CD40L MAb does not inhibit killing of intracellular M. tuberculosis by antigen-specific CD4+ T cells. CD4+ T cells were isolated from PBMC that had been stimulated for 7 days by incubation with avirulent M. tuberculosis H37Ra. As illustrated, the addition of these cells in a 5:1 ratio relative to infected MN resulted in a significant reduction of intracellular M. tuberculosis at 1, 4, and 7 days of coculture. The addition of 5c8 in concentrations of 10 and 100 μg/ml had no effect on CD4+ T-cell-mediated killing of H37Rv.

Techniques: Isolation, Incubation, Infection

Review

Structured Review

Images

1) Product Images from "CD40L induces functional tunneling nanotube networks exclusively in dendritic cells programmed by mediators of type-1 immunity"

Similar Products

Image Search Results