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