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: Journal of Inflammation (London, England)

Article Title: Tissue-plasminogen activator effects on the phenotype of splenic myeloid cells in acute inflammation

doi: 10.1186/s12950-024-00375-0

Figure Lengend Snippet: References of flow cytometry antibodies used for immunophenotyping

Article Snippet: CD80 , REA983 , APC-Vio770 , 130–116-463 , human IgG1/REA293 , Miltenyi Biotec.

Techniques: Flow Cytometry, Clone Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Novel Pt(IV) complex OAP2 induces STING activation and pyroptosis via mitochondrial membrane remodeling for synergistic chemo-immunotherapy

doi: 10.1016/j.apsb.2023.11.032

Figure Lengend Snippet: The immune activation effect of OAP2 in vivo . (A–B) Analysis of mature DC cells in mouse spleen. Expression of CD80 and CD86 quantitatively detected by flow cytometry (A), and statistical analysis (B). (C–D) Analysis of mature DC cells in mouse tumor site. Expression of CD80 and CD86 quantitatively detected by flow cytometry (C), and statistical analysis (D). (E–F) Analysis of cytotoxic T cells in mouse tumor. Expression of CD4 and CD8 quantitatively detected by flow cytometry (E), and statistical analysis (F). (G–N) Tumor tissue cytokine expression content measured by ELISA ( n = 3).

Article Snippet: The following antibodies and kits were used in the flow cytometry and ELISA analyses: CoraLite® Plus 488 Anti-Mouse CD4, CoraLite®568 Anti-Mouse CD3, APC Anti-Mouse CD86, FITC Plus Anti-Mouse CD11c, and PE Anti-Mouse CD80 (B7-1) were purchased from Proteintech.

Techniques: Activation Assay, In Vivo, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay