Fig 1: ELISA-based determination of plasma sPD-L1 and sPD-L2 from the blood of patients with endometrial cancers: Average sPD-L1 (A) and sPD-L2 (B) from the blood (plasma) of age-matched normal subjects (solid black circles) and patients with endometrial cancers (peach triangles) are presented. Data from patients with ovarian cancers were included as an internal control, as indicated by teal triangles. The difference between the means is presented in the estimation plot. The p value (unpaired t-test) is <0.0001 for sPD-L1 (n = 19 for normal subjects and n = 11 for cancer patients). The p-value (unpaired t-test) is <0.05 for sPD-L2 (n = 20 for normal subjects and n = 11 for cancer patients). The rightmost column in the estimation plot represents the difference between the means of the two groups (in pg/mL units). Hence in Figure 7A, the plot represents the difference between Patient (Endo + Ovary) Plasma and Normal Plasma, and it is a positive value as the expression in the Patient (Endo + Ovary) Plasma was higher than that in Normal Plasma PD-L1. In (B), the plot represents the difference between Patient (Endo + Ovary) Plasma and Normal Plasma Average from 1/50, and it is a negative value as the expression in the patient (Endo + Ovary) plasma was lower than that in Normal Plasma PD-L2 Average from 1/50. Plasma sPD-L1 and sPD-L2 expression by ELISA and their ratios in the blood of patients with endometrial cancers are plotted individually (C).
Fig 2: PD-L1/PD-L2 and PD-1 status of epithelial cells and cells of the stromal microenvironment, SME (lymphocytes, L; macrophages, M;) from tumor-adjacent normal tissues: (A–F): Expression of PD-L1/PD-L2 and PD-1 by IHC of FFPE sections from epithelial cells of tumor-adjacent normal tissues. The expression of PD-L1/PD-L2 and PD-1 in mesenchymal cells of the stromal microenvironment, SME (lymphocytes, macrophages, and blood vessels) from FFPE sections by IHC is presented. The green square/rectangle is an epithelial cell; the green circle is a lymphocyte (L); the green freeform is a macrophage (M); the green pentagon represents mesenchyme. Insets show images of the selected cells at higher magnifications. Bars represent 20 µM.
Fig 3: PD-L1/PD-L2 and PD-1 status of tumor cells and cells of the tumor microenvironment, TME (lymphocytes, L; macrophages, M; and blood vessels, BV) from tumor tissues: (A): Plan of the study for obtaining PD-L1/PD-L2 and PD-1 status of tumor cells’ tumor-adjacent normal tissue, cells of the tumor microenvironment, TME (lymphocytes, L; macrophages, M; and blood vessels, BV), and cells of the stromal microenvironment, SME, of patients with endometrial cancers. (B–D): Expression of PD-L1/PD-L2 and PD-1 in tumor cells from IHC of FFPE sections. (E–I): Expression of PD-L1/PD-L2 and PD-1 in cells of the TME (lymphocytes, macrophages, and blood vessels) from FFPE of tumor tissues. The red square is a tumor cell; the red circle is a lymphocyte (L); the red arrow is a blood vessel (BV); the red freeform is a macrophage (M). Insets show images of the selected cells at higher magnifications. Bars represent 20 µM.
Fig 4: Expression of PD-L1 and PD-L2 mRNA in paired CAFs in primary culture from tumor and tumor-adjacent normal samples obtained from patients with endometrial cancers: PD-L1 (A) and PD-L2 (B) mRNA expression (relative ratios to GAPDH) of NCAFs and TCAFs were carried out by qRT-PCR. HUF and tumor cell lines were used as controls. Melting curves for the respective mRNAs and GAPDH are presented as insets. A heatmap of PD-L1 and PD-L2 mRNA expression is presented as ratios of TCAFs/NCAFs pairs (C) of different patients with endometrial cancers. The five-rating icon sets (bars) with a three-color scale (light blue as a minimum, yellow as the midpoint, and violet as the maximum) are used.
Fig 5: Conditional formatting by five-rating icon sets of paired T-TME (A) and N-SME (B) staining for PD-L1, PD-L2, and PD-1 by IHC on FFPE sections from Day0 that were used for sorting by grades 1, 2 and 3 of the disease. A. Conditional formatting of PD-L1, PD-L2, and PD-1 stains by five-rating icon sets of tumor cells (numbers in violet font represent Tumor Proportion Score, TPS (<%) in dark-orange filled rows) and cells of the TME (numbers in green font represent % of positive cells in the TME, including lymphocytes, macrophages, and around blood vessels, in lighter orange rows). Semi-quantification of TILs is represented in the figure. * represents lymphovascular invasion, and ** represents poorly differentiated regions of the tumor. (B). Conditional formatting of PD-L1, PD-L2, and PD-1 stains by five-rating icon sets of tumor-adjacent normal tissues (numbers in blue font represent positive epithelial cells (%) in dark green filled rows) and SME (numbers in black font represent % of positive cells in SME, including lymphocytes, macrophages, and around blood vessels, in lighter green rows).
Supplier Page from Abcam for Human PD-L2 ELISA Kit