pe anti human cd155  (Miltenyi Biotec)

Journal: Frontiers in Immunology

Article Title: Enhancing anti-tumor immunity through co-blocking PD-L1 and TIGIT by facilitating tumor-directed responses and additional VEGF inhibition

doi: 10.3389/fimmu.2025.1746155

Figure Lengend Snippet: Pattern of expression of PD-L1 and CD155 (PVR) and its correlation to blocking and immune landscape. (A) FACS plots show expression pattern of PD-L1 and CD155 by HT-1080 cells. (B) CTV-labelled PBMCs were cultured with mitomycin-treated HT-1080 cells in 96-well plates with indicated Abs at equal molar concentrations. Histograms show the numbers of proliferated cells under stimulated conditions. **** p < 0.0001 by one-way ANOVA multiple-comparisons test. (C–H). High-Content Analysis of human lung adenocarcinoma microenvironment. Multiplex immunofluorescence and quantitative analysis were used to profile the tumor (PanCK + ) and stromal (PanCK - ) compartments. (C) Representative High-Content analysis images illustrating the immune landscape in adenocarcinoma tissue. (D) Quantification of the percentage of cells expressing PD-L1 within the PanCK + tumor and PanCK - stromal compartments. Lines connect paired tumor and stroma samples from the same patient. The mean difference between compartments is plotted on the right axis for each graph. PD-L1 expression was then further expressed as Tumor Proportion Score (TPS), calculated as the ratio of PanCK + PD-L1 + cells to PanCK + tumor cells, and a Combined Positive Score (CPS), calculated as the ratio of all PD-L1 + cells to PanCK + tumor cells. (E) Quantification of the percentage of cells expressing CD155 within the PanCK + tumor and PanCK - stromal compartments. Lines connect paired tumor and stroma samples from the same patient. The mean difference between compartments is plotted on the right axis for each graph. (F) Co-expression analysis of PD-L1 and CD155 on PanCK + tumor cells. Each dot represents a patient sample. Dashed lines indicate the median density of the cohort for each marker, stratifying tumors into four subsets. (G) Graphs show Frequencies of CD4 + and CD8 + T cells, calculated as a percentage of total cells within the tissue; Frequencies of PD-1, TIGIT, and CD226 expression on CD4 + and CD8 + T cells, shown as a percentage of the parent T-cell population; and Linear regression analysis showing a significant positive correlation between the density (cells/mm 2 ) of TIGIT + and CD226 + cells within the CD4 + (left) and CD8 + (right) T cell populations. R 2 and P values are shown. (H) Correlation analysis between the density of total PD-L1 + cells (left) or total CD155 + cells (right) and the density of CD4 + (red line) or CD8 + (blue line) T cells.

Article Snippet: Cells were activated with PHA-M (Yeasen, Cat#: 40110ES08, lot: P2107890; 0.95 μg/mL final concentration) and co-treated with soluble CD155 (Sino Biological, Cat#10109-H02H; 8 μg/mL final concentration).

Techniques: Expressing, Blocking Assay, Cell Culture, High Content Screening, Multiplex Assay, Immunofluorescence, Marker

Journal: Science Advances

Article Title: HIV-1 manipulates CD96 on CD4 + T cells to subvert antiviral immunity

doi: 10.1126/sciadv.adx7485

Figure Lengend Snippet: Primary CD4 + T cells were infected with HIV-1 NL4-3–IRES–eGFP reporter virus containing the indicated deletions in vpr , nef , and vpu . At 48 hours postinfection, cells were harvested and stained for the indicated cell surface receptors with specific antibodies and analyzed by flow cytometry. ( A ) Representative dot plots of HIV-1–infected primary CD4 + T cells that were stained for CD96 cell surface expression. GFP was used to identify HIV-1–infected cells (green box), and modulation was directly compared to uninfected cells (blue box). ( B ) CD96, ( C ) CD226, ( D ) CD155, and ( E ) NTB-A MFIs were used to calculate the respective surface expression of HIV-1–infected cells compared to uninfected cells. ( F ) Infection of primary CD4 + T cells with HIV-1 NL4-3 and two HIV-1 strains isolated from PLWH during the chronic stage (CH058 and CH077) harboring inactivating mutations in vpu and nef and assessment of cell surface CD96 levels via flow cytometry at 3 days postinfection. Values from (B) nine (NL4-3, Nef − , Vpu − , and N − U − ), six (Vpr − ), and four (N − U − R − ) and for [(C) to (E)] four and two (N − U − R − ) independent experiments were plotted (means ± SD). (F) Data from three to five independent donors (means ± SD), statistical differences were assessed using a one-way analysis of variance (ANOVA) with Dunnett’s multiple comparison test. **** P < 0.0001, *** P < 0.001, ** P < 0.01, and * P < 0.05.

Article Snippet: PE anti-human CD155 , Flow cytometry , 1:50 in 1% FBS in PBS , Miltenyi Biotec , 130-105-905.

Techniques: Infection, Virus, Staining, Flow Cytometry, Expressing, Isolation, Comparison

Journal: Science Advances

Article Title: HIV-1 manipulates CD96 on CD4 + T cells to subvert antiviral immunity

doi: 10.1126/sciadv.adx7485

Figure Lengend Snippet: Primary CD4 + T cells were IL-2/PHA stimulated for 3 days and left untreated ( A ) or subjected to CRISPR-Cas9–mediated CD96 KO ( B ). Twenty-four hours later, cells were restimulated with anti-CD3/anti-CD28, and, 48 hours later, cells were analyzed for cytokine secretion and costained for CD96. Cytokine secretion of (A) CD96 Hi cells was compared to CD96 Lo cells and (B) that of CD96 KO cells to scr-RNA control cells ( n = 8, means ± SEM, two-way ANOVA with Sidak’s multiple comparison). ** P < 0.01 and * P < 0.05. ( C ) An hCD96-GFP fusion protein is predominantly localized on the cell membrane of medaka thymic T cells (movie S1) compared with GFP-only–expressing controls (movie S2). Scale bars, 30 μm. Average cell speed ( D ) and straightness ( E ) of GFP + cells in the thymus. N = total number of cells examined from >3 samples per condition (means ± SEM, two-tailed Mann-Whitney test). **** P < 0.0001. The asterisk (*) marks an autofluorescent pigment cell cluster. ( F ) Primary CD4 + T cells prestimulated with CD3/CD28 for 3 days were infected with HIV-1 NL4-3–IRES–eGFP or the N − /U − variant. Forty-eight hours postinfection, cells were seeded onto 96-well plates coated with CD96 ligands Nectin-1, CD155, and anti-CD96 or an isotype control. After 45 min at 37°C, nonadherent and adherent cells were collected separately and quantified by flow cytometry. Adhesion was calculated as the percentage of adherent cells relative to the total number of cells ( n = 5, means ± SEM, paired two-way ANOVA with Fisher’s least significant difference). ** P < 0.01 and * P < 0.05.

Article Snippet: PE anti-human CD155 , Flow cytometry , 1:50 in 1% FBS in PBS , Miltenyi Biotec , 130-105-905.

Techniques: CRISPR, Control, Comparison, Membrane, Expressing, Two Tailed Test, MANN-WHITNEY, Infection, Variant Assay, Flow Cytometry

Journal: Science Advances

Article Title: HIV-1 manipulates CD96 on CD4 + T cells to subvert antiviral immunity

doi: 10.1126/sciadv.adx7485

Figure Lengend Snippet: ( A ) PBMCs were stimulated with HCMV pp65 peptide and costimulated either alone or in combination with CD155 or anti-CD96 (each 5 μg/ml) and stained for IFN-γ 21 hours later. IFN-γ secretion was calculated as fold change relative to pp65 stimulation only (mock). ( n = 10, Kruskal-Wallis test with uncorrected Dunn’s test). * P < 0.05 and ** P < 0.01. Depicted are primary data from two donors. ( B ) Resting primary CD4 + T cells were infected with HIV-1 NL4-3–IRES–eGFP or the N − /U − variant and stimulated at 6 hours postinfection with CD3/CD28 antibodies. Two days postinfection, cells were restimulated with CD3/CD28 antibodies either alone or in combination with anti-CD96 or an isotype control. Three days later, IFN-γ secretion was assessed via flow cytometry ( n = 5, paired one-tailed t test). ( C ) Primary CD4 + T cells were stimulated with CD3/CD28 antibodies (0.5 μg/ml each) and mock treated or costimulated with coated CD96 antibody (2.5 μg/ml) or isotype (2.5 μg/ml). Three days poststimulation, supernatants were harvested for a cytokine array ( n = 3, one representative array shown). GM-CSF, granulocyte-macrophage colony-stimulating factor. ( D ) Heatmap illustrating relative abundance of analyzed cytokines normalized to the respective controls. [Mean values, n = 3; n = 2 for IL-6, macrophage colony-stimulating factor (M-CSF), and fibroblast growth factor 6 (FGF-6)]. Only cytokines with >2% relative abundance in at least one treatment condition were included. LIF, leukemia inhibitory factor; BDNF, brain-derived neurotrophic factor; SCF, stem cell factor; MIF, migration inhibition factor; EGF, epidermal growth factor; GDNF, glial cell line–derived neurotrophic factor. ( E ) X-fold change in cytokine secretion between CD96 antibody versus isotype-treated CD4 + T cells was calculated, and the waterfall plot depicts changes of >2 ( n = 3, ±SD, multiple unpaired t test with individual P values). ( F ) Pathway analyses using Enrichr (Kyoto Encyclopedia of Genes and Genomes 2021 database) and as input the nine significantly regulated genes from (E). Bar length and brightness correlates with significance ( q > 0.0001). For “IL-17 signaling,” key cytokines from (E) are indicated.

Article Snippet: PE anti-human CD155 , Flow cytometry , 1:50 in 1% FBS in PBS , Miltenyi Biotec , 130-105-905.

Techniques: Staining, Infection, Variant Assay, Control, Flow Cytometry, One-tailed Test, Derivative Assay, Migration, Inhibition

Journal: Diagnostics

Article Title: Prognostic and Predictive Significance of B7-H3 and CD155 Expression in Gastric Cancer Patients

doi: 10.3390/diagnostics15212695

Figure Lengend Snippet: Fold change expression levels of B7-H3 and CD155 across individual gastric cancer cases. A fold change greater than 1 was interpreted as positive or upregulated gene expression, where fold regulation was considered equivalent to the fold change value. A fold change greater than 2 was classified as moderate to high upregulation in gene expression.

Article Snippet: For B7-H3, a recombinant rabbit monoclonal IgG antibody (HUABIO, Woburn, MA, USA), clone: HA721245; dilution 1:5000) was applied, while for CD155, a recombinant rabbit monoclonal IgG antibody (Proteintech, Rosemont, IL, USA, clone: 241405F5; dilution 1:1500) was used.

Techniques: Expressing, Gene Expression

Journal: Diagnostics

Article Title: Prognostic and Predictive Significance of B7-H3 and CD155 Expression in Gastric Cancer Patients

doi: 10.3390/diagnostics15212695

Figure Lengend Snippet: Comparison of B7-H3 and CD155 fold-Change Expression According to Metastatic Status (M0 vs. M1). Fold change was calculated by dividing the normalized gene expression in the test sample by the normalized gene expression in the control sample. Fold regulation represents the biologically relevant direction and magnitude of change derived from the fold change value.

Article Snippet: For B7-H3, a recombinant rabbit monoclonal IgG antibody (HUABIO, Woburn, MA, USA), clone: HA721245; dilution 1:5000) was applied, while for CD155, a recombinant rabbit monoclonal IgG antibody (Proteintech, Rosemont, IL, USA, clone: 241405F5; dilution 1:1500) was used.

Techniques: Comparison, Expressing, Gene Expression, Control, Derivative Assay