Fig 1: Immunohistochemical staining against PD-1, PD-L1 and PD-L2. Representative images of FFPE samples immunostained with PD-1 (a-d), PD-L1 (e-h) and (i-l), original magnification: 200×. A, E and I represented negative expression of PD-1, PD-L1 and PD-L2, respectively. B-D, F-H and J-L represented different positive intensities (1+, 2+ and 3+) of PD-1, PD-L1 and PD-L2, respectively
Fig 2: The representative images of immune signatures. (A) The intratumor infiltration of CD45+ cells (CD45 is a general lymphocyte marker), CD8+ cells (CD8 is a cytotoxic T lymphocyte marker), CD45RO+ cells (CD45RO is a memory T lymphocyte marker), PD1+ cells (PD1 is an immunosuppressive lymphocyte marker) within TME. (B) PD-L1 expression on tumor cells and intraepithelial TILs within TME. (C) PD-L2 was partially expressed in intraepithelial TILs within TME. Scale bar = 50 µm.
Fig 3: High tumor PD-L2 is remarkably associated with favorable 5-year OS in patients who receive postoperative chemotherapy. (A) High tumor PD-L2 was associated with better 5-year OS in advanced stage colon carcinoma patients (n = 228, p = 0.0323). CT: chemotherapy. (B) High tumor PD-L2 levels were remarkably associated with favorable 5-year OS in advanced stage colon carcinoma patients who received postoperative chemotherapy (n = 405, p < 0.001). (C) Patients with high tumor PD-L2 level was significantly associated with better 5-year OS in stage III colon carcinoma patients who received adjuvant chemotherapy. CT: chemotherapy (n = 276, p = 0.0492). (D) The multivariate COX regression model showed tumor PD-L2 is an independent factor in 5-year OS of stage III COAD patients who received post-operative chemotherapy. The hazard ratios (HR) and 95% confidence interval (CI) of different factors in the COX regression model. (E) Patients with high tumor PD-L2 level were tendency to associate with better 5-year OS in stage IV colon carcinoma patients who received palliative chemotherapy. CT: chemotherapy (n = 129, p = 0.0561). (F) The multivariate COX regression model suggested tumor PD-L2 is an independent factor in 5-year OS of stage IV COAD patients who received post-operative palliative chemotherapy. The HR and 95% CI of different factors in the COX regression model.
Fig 4: The association of tumor PD-L2 level and CD8+ TIL and CD45RO+ TIL density with 5-year OS in advanced stage colon carcinoma. (A) Advanced stage colon carcinoma patients with high infiltration of intratumoral CD8+ TIL within the TME had a better 5-year OS (n = 630, p = 0.0005). (B) The tumor PD-L2 level was not associated with 5-year OS in patients with a high density of CD8+ TILs within the TME (n = 138, p = 0.167). (C) The tumor PD-L2 level was associated with 5-year OS in patients with a low density of CD8+ TILs within the TME (n = 492, p < 0.001). (D) Advanced stage colon carcinoma patients with a high density of CD45RO+ TILs within the TME had a better 5-year OS (n = 627, p < 0.001). (E) The tumor PD-L2 level was not associated with 5-year OS in patients with a high density of CD45RO+ TILs within the TME (n = 173, p = 0.3785). (F) The tumor PD-L2 level was associated with 5-year OS in patients with a low density of CD45RO+ TILs within the TME (n = 454, p < 0.001).
Fig 5: Representative pictures of lymphocyte infiltration in triple-negative breast cancer core biopsy showing immunohistochemical staining of high CD3+, CD4+, CD8+, CD14+, CD68+, CD117+, FOXP3+, PD-1+, PD-L1+, and PD-L2+. The specimens are imaged at ×20 magnification with a 100-µm scale on each one.
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