Journal: Immunity

Article Title: PD-L1:CD80 Cis -Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways

doi: 10.1016/j.immuni.2019.11.003

Figure Lengend Snippet: (A) Representative flow-cytometry histograms of PD-1-huFc staining of the indicated types of Raji cells. Bound PD-1-huFc was labeled by AF647 anti-human IgG Fc, the mean fluorescence intensity (MFI) of which was plotted against PD-1-huFc concentration (means ± SEM, n = 3). In gray is the staining of Raji (CD80−PD-L1-mCherry+) by isolated huFc domain.

Article Snippet: For , SLB was functionalized by a mixture of 5 nM pMHC-I-His, 2 nM mouse ICAM–His (Sino Biological, 50440-M08H) and either 3 nM mouse PD-L1–His (Sino Biological, 50010-M08H), 3 nM mouse PD-L1–His plus 9 nM mouse CD80–His (Sino Biological, 50446-M08H), or 3 nM mouse PD-L1–His plus 9 nM mouse CD86–His (Sino Biological, 50068-M08H).

Techniques: Flow Cytometry, Staining, Labeling, Fluorescence, Concentration Assay, Isolation

Journal: Immunity

Article Title: PD-L1:CD80 Cis -Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways

doi: 10.1016/j.immuni.2019.11.003

Figure Lengend Snippet: (A) Representative flow-cytometry histograms of CD28-huFc staining of the indicated types of Raji cells. Bound CD28-huFc was labeled by AF647 anti-human IgG Fc, the MFI of which was plotted against (CD28-huFc. Shown in gray are Raji (CD80+CD86−) cells stained by isolated huFc domain. Means ± SEM, n = 3.

Article Snippet: For , SLB was functionalized by a mixture of 5 nM pMHC-I-His, 2 nM mouse ICAM–His (Sino Biological, 50440-M08H) and either 3 nM mouse PD-L1–His (Sino Biological, 50010-M08H), 3 nM mouse PD-L1–His plus 9 nM mouse CD80–His (Sino Biological, 50446-M08H), or 3 nM mouse PD-L1–His plus 9 nM mouse CD86–His (Sino Biological, 50068-M08H).

Techniques: Flow Cytometry, Staining, Labeling, Isolation

Journal: Immunity

Article Title: PD-L1:CD80 Cis -Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways

doi: 10.1016/j.immuni.2019.11.003

Figure Lengend Snippet: (A) A Jurkat-Raji co-culture assay analyzing how PD-L1 interferes with CTLA-4-mediated CD80 depletion. Cartoons on the left depict the co-cultured cells. On the immediate right are representative flow-cytometry histograms of CD80 expression (anti-CD80 allophycocyanin) on Raji cells before (0 h) and after co-culture (0.5 h). Further on the right are representative confocal images for the Jurkat-Raji conjugate (scale bars, 10 μm). Rightmost is a bar graph showing CD80 MFI of Raji at 0.5 h, normalized to CD80 MFI at 0 h (mean ± SEM, n = 4).

Article Snippet: For , SLB was functionalized by a mixture of 5 nM pMHC-I-His, 2 nM mouse ICAM–His (Sino Biological, 50440-M08H) and either 3 nM mouse PD-L1–His (Sino Biological, 50010-M08H), 3 nM mouse PD-L1–His plus 9 nM mouse CD80–His (Sino Biological, 50446-M08H), or 3 nM mouse PD-L1–His plus 9 nM mouse CD86–His (Sino Biological, 50068-M08H).

Techniques: Co-culture Assay, Cell Culture, Flow Cytometry, Expressing, Co-Culture Assay

Journal: Immunity

Article Title: PD-L1:CD80 Cis -Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways

doi: 10.1016/j.immuni.2019.11.003

Figure Lengend Snippet: (A) Representative TIRF images of PD-L1 LUVs captured by PD-1 SLB, CD80 SLB, or CD86 SLB; each LUV is registered as a green spot. Bar graph summarizes the fluorescence intensity (FI) of the LUV channel under indicated conditions, normalized to the intensity of the condition with PD-1 SLBs. Data are means ± SEM, n = 3. Scale bars, 5 μm.

Article Snippet: For , SLB was functionalized by a mixture of 5 nM pMHC-I-His, 2 nM mouse ICAM–His (Sino Biological, 50440-M08H) and either 3 nM mouse PD-L1–His (Sino Biological, 50010-M08H), 3 nM mouse PD-L1–His plus 9 nM mouse CD80–His (Sino Biological, 50446-M08H), or 3 nM mouse PD-L1–His plus 9 nM mouse CD86–His (Sino Biological, 50068-M08H).

Techniques: Fluorescence

Journal: Immunity

Article Title: PD-L1:CD80 Cis -Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways

doi: 10.1016/j.immuni.2019.11.003

Figure Lengend Snippet: (A) Representative flow-cytometry histograms of CTLA-4-huFc staining of the indicated types of Raji cells. Bound CTLA-4-huFc was labeled by AF647 anti-human IgG Fc, the MFI of which was plotted against (CTLA-4-huFc). Shown in gray are Raji (CD80+CD86−) cells stained by isolated huFc domain. Means ± SEM, n ≥ 3.

Article Snippet: For , SLB was functionalized by a mixture of 5 nM pMHC-I-His, 2 nM mouse ICAM–His (Sino Biological, 50440-M08H) and either 3 nM mouse PD-L1–His (Sino Biological, 50010-M08H), 3 nM mouse PD-L1–His plus 9 nM mouse CD80–His (Sino Biological, 50446-M08H), or 3 nM mouse PD-L1–His plus 9 nM mouse CD86–His (Sino Biological, 50068-M08H).

Techniques: Flow Cytometry, Staining, Labeling, Isolation

Journal: Immunity

Article Title: PD-L1:CD80 Cis -Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways

doi: 10.1016/j.immuni.2019.11.003

Figure Lengend Snippet: (A) On the left are flow-cytometry histograms of CD80 and CD86 surface levels on DCs and macrophages (Macs) isolated from tumor tissues of 4T1 implanted BALB/C female mice treated with anti-PD-L1 (magenta traces), anti-PD-1 (cyan traces), or control IgG (black traces). On the right are bar graphs summarizing the MFI of CD80 and CD86 staining under the indicated conditions. Data are shown as mean ± SEM, n ≥ 3 mice.

Article Snippet: For , SLB was functionalized by a mixture of 5 nM pMHC-I-His, 2 nM mouse ICAM–His (Sino Biological, 50440-M08H) and either 3 nM mouse PD-L1–His (Sino Biological, 50010-M08H), 3 nM mouse PD-L1–His plus 9 nM mouse CD80–His (Sino Biological, 50446-M08H), or 3 nM mouse PD-L1–His plus 9 nM mouse CD86–His (Sino Biological, 50068-M08H).

Techniques: Flow Cytometry, Isolation, Staining

Journal: Immunity

Article Title: PD-L1:CD80 Cis -Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways

doi: 10.1016/j.immuni.2019.11.003

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: For , SLB was functionalized by a mixture of 5 nM pMHC-I-His, 2 nM mouse ICAM–His (Sino Biological, 50440-M08H) and either 3 nM mouse PD-L1–His (Sino Biological, 50010-M08H), 3 nM mouse PD-L1–His plus 9 nM mouse CD80–His (Sino Biological, 50446-M08H), or 3 nM mouse PD-L1–His plus 9 nM mouse CD86–His (Sino Biological, 50068-M08H).

Techniques: Recombinant, Isolation, Enzyme-linked Immunosorbent Assay, Cell Isolation, Software, Imaging

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Characterization of immune cell subtypes in three commonly used mouse strains reveals gender and strain-specific variations

doi: 10.1038/s41374-018-0137-1

Figure Lengend Snippet: Details of antibodies used in the study

Article Snippet: CD80 , APC , Miltenyi , 16–10A1 , 130–102-584 , 3.75 pL.

Techniques: Concentration Assay

Journal:

Article Title: CTLA-4 regulates allergen response by modulating GATA-3 protein level per cell

doi: 10.1111/j.1365-2567.2007.02537.x

Figure Lengend Snippet: Effects of in vitro CTLA-4 blockade on GATA-3 protein level/cell and frequency of IL-4-producing cells. Naive CD4+ T cells purified from the spleens of normal mice were induced to differentiate with anti-CD3 mAb, a recombinant form of CD80-Fc, IL-4 and anti-IFN-γ mAb for 3 days. Anti-CTLA-4 mAb was added in soluble form at increasing concentrations (0·1–10 μg/ml). (a) Mean fluorescence intensity for GATA-3 staining, (b) percentage of IL-4+ cells.

Article Snippet: 18 CD4 cell culture For in vitro experiments, splenic naive CD4 cells were purified from non-immunized BALB/c mice by immuno-magnetic cell sorting as previously described 19 and cultured in 24-well plates precoated with anti-CD3ε mAb (clone 145-2C11; 10 μg/ml) and/or recombinant mouse B7-1(CD80)/Fc chimeric protein (R & D Systems, cat. 740-B1; 0·33 μg/ml).

Techniques: In Vitro, Purification, Recombinant, Fluorescence, Staining

Journal: Pediatric nephrology (Berlin, Germany)

Article Title: Glomerular endothelial cells and podocytes can express CD80 in patients with minimal change disease during relapse

doi: 10.1007/s00467-020-04541-3

Figure Lengend Snippet: (A) Immunostaining with a mouse monoclonal anti human CD80 (red) was performed on paraffin embedded sections from patients with MCD and FSGS. Positive and negative controls are shown (upper panel). Upregulation of glomerular CD80 was noted in most of MCD but few FSGS patients (lower panel). (B) CD80 (green, upper panel) colocalized with caveolin 1 in an endothelial specific pattern (arrows). To less extent, CD80 (red, lower panel) colocalized with nephrin (arrowheads). (C) Specificity of immunostaining was confirmed by silver staining. No signal detected in kidney tissue sample from same MCD patient in relapse when primary antibody was omitted. (D) Electron microscopy (EM) from kidney tissue from patient with MCD is shown. CD80 colocalized with glomerular endothelial cells and rarely podocytes. Original EM images captured at 6000x (far right) and 30000x. Podocyte (P), capillary lumen (CL), endothelial cell (EC), urinary space (U), glomerular basement membrane (GBM). Scale bars: 100 μm.

Article Snippet: *Immunofluorescence on fresh tissue (when available) using a polyclonal CD80 goat antibody (R&D systems) *Immunoperoxidase staining on paraffin-embedded tissue using a mouse monoclonal CD80 antibody (R&D systems) *Adults *CD80+ in interstitial inflammatory cells and injured epithelial cells Lee 23 28 Primary MCD, relapse 0/28 podocyte CD80+ 21/28 tubular CD80+ *Immunohistochemistry on paraffin-embedded tissue using a mouse monoclonal CD80 antibody (R&D systems) *Adults Open in a separate window CD80 expression in kidney tissue from patients with idiopathic nephrotic syndrome The first major finding in this study was to identify CD80 on the surface of glomerular endothelial cells as well as podocytes in a subset of MCD patients.

Techniques: Immunostaining, Silver Staining, Electron Microscopy, Membrane

Journal: Pediatric nephrology (Berlin, Germany)

Article Title: Glomerular endothelial cells and podocytes can express CD80 in patients with minimal change disease during relapse

doi: 10.1007/s00467-020-04541-3

Figure Lengend Snippet: (A) Immunostaining was performed on paraffin embedded sections from mice treated with PBS (controls) and LPS. Positive and negative controls are shown. CD80 was upregulated in glomeruli following LPS. Scattered infiltrating inflammatory and/or parietal cells were also observed after LPS (upper panel, arrowheads). (B) By immunostaining, CD80 (green) colocalized to some extent with synaptopodin (podocyte marker, red) and lined the capillary lumen co-localizing with ICAM1 (red), used as marker of endothelial cell activation. (C) Using in situ hybridization, we confirmed the presence of CD80 RNA molecules in spleen (positive control) and glomeruli from LPS mice (24 hours). PPIB (housekeeping gene coding for peptidylprolyl isomerase B) probe was also used as positive control. Omission of CD80 probes on spleen and kidney were used as negative controls. (D) CD80 and ICAM1 mRNA were upregulated in LPS-treated mice, compared to controls, as early as 6 hours following LPS. Scale bars: 100 μm.

Article Snippet: *Immunofluorescence on fresh tissue (when available) using a polyclonal CD80 goat antibody (R&D systems) *Immunoperoxidase staining on paraffin-embedded tissue using a mouse monoclonal CD80 antibody (R&D systems) *Adults *CD80+ in interstitial inflammatory cells and injured epithelial cells Lee 23 28 Primary MCD, relapse 0/28 podocyte CD80+ 21/28 tubular CD80+ *Immunohistochemistry on paraffin-embedded tissue using a mouse monoclonal CD80 antibody (R&D systems) *Adults Open in a separate window CD80 expression in kidney tissue from patients with idiopathic nephrotic syndrome The first major finding in this study was to identify CD80 on the surface of glomerular endothelial cells as well as podocytes in a subset of MCD patients.

Techniques: Immunostaining, Marker, Activation Assay, In Situ Hybridization, Positive Control

Journal: Pediatric nephrology (Berlin, Germany)

Article Title: Glomerular endothelial cells and podocytes can express CD80 in patients with minimal change disease during relapse

doi: 10.1007/s00467-020-04541-3

Figure Lengend Snippet: CD80 expression in kidney tissue from patients with idiopathic nephrotic syndrome

Article Snippet: *Immunofluorescence on fresh tissue (when available) using a polyclonal CD80 goat antibody (R&D systems) *Immunoperoxidase staining on paraffin-embedded tissue using a mouse monoclonal CD80 antibody (R&D systems) *Adults *CD80+ in interstitial inflammatory cells and injured epithelial cells Lee 23 28 Primary MCD, relapse 0/28 podocyte CD80+ 21/28 tubular CD80+ *Immunohistochemistry on paraffin-embedded tissue using a mouse monoclonal CD80 antibody (R&D systems) *Adults Open in a separate window CD80 expression in kidney tissue from patients with idiopathic nephrotic syndrome The first major finding in this study was to identify CD80 on the surface of glomerular endothelial cells as well as podocytes in a subset of MCD patients.

Techniques: Expressing, Immunohistochemistry, Immunoperoxidase Staining, Immunofluorescence

Journal: Veterinary Research

Article Title: Interactions between natural killer cells and dendritic cells favour T helper1-type responses to BCG in calves

doi: 10.1186/s13567-016-0367-4

Figure Lengend Snippet: BCG is taken up by bovine monocyte-derived DCs resulting in increased the expression of MHC class II, CD40 and CD80 and production of the Th1 polarising cytokine IL-12. Monocyte-derived DCs were cultured for 3 days and infected with FITC-labelled BCG (MOI 5) for 42 h. FACS plots from one representative animal ( A ) illustrate the percentage uptake of BCG-FITC by uninfected DCs (blue histogram) and BCG-infected DCs (pink histogram). Gates were set using uninfected DCs. Pooled data from four calves ( A ) illustrate the average percentage uptake of BCG-FITC ± SD by uninfected DCs (lighter bar) and BCG-infected DCs (darker bar). Uninfected and BCG-infected DCs were labelled with mAbs for MHC class II, CD40 and CD80 and analysed by flow cytometry. FACS plots from one representative animal show the expression of MHC class II ( B ), CD40 ( C ) and CD80 ( D ) by uninfected (blue histograms) and BCG-infected DCs (pink histograms). Positive cells were identified based on FMO controls (green histograms). Pooled data from four calves indicates the average MFI ± SD of MHC class II ( B ), CD40 ( C ) and CD80 ( D ) expression by uninfected (lighter bars) and BCG-infected DCs (darker bars). Supernatants were retrieved from uninfected and BCG-infected DCs and assayed for IL-12 production by ELISA. Pooled data from five calves represent the average levels ± SD of IL-12 BU/mL produced by uninfected (lighter bars) and BCG-infected DCs (darker bars) ( E ). Data were normally distributed ( p > 0.05) and significance between uninfected and BCG-infected DCs was assessed using a 2-sample t test. p < 0.05*, p < 0.001***.

Article Snippet: Mouse anti-bovine MHC class II-PE (CC158, IgG2a, Bio-Rad AbD Serotec), mouse anti-bovine CD40-FITC (IL-A156, IgG1, Bio-Rad AbD Serotec) and mouse anti-bovine CD80 (IL-A159, Bio-Rad AbD Serotec) indirectly conjugated to goat anti-mouse IgG1-AF647 (Life Technologies, UK) were used to determine DC expression of MHC class II, CD40 and CD80 respectively.

Techniques: Derivative Assay, Expressing, Cell Culture, Infection, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Produced