Fig 1: LMP1 and PD-L1 expression in NPC tissue samples. The specimens were immunostained with LMP1 and PD-L1 antibody. Brown staining indicates positive cell membrane expression of LMP1 (A) and PD-L1 (B). Dual fluorescence immunostaining of LMP1 (C) and PD-L1 (D), and DAPI (E) in NPC tissue samples. (F) Merged image of LMP1 and PD-L1. White arrowheads indicate the colocalization of LMP1 and PD-L1. Original magnifications (A,B): ×100, inset figures: ×400, scale bar: 100 µm. Original magnifications (C–F): ×600.
Fig 2: Elevated serum sPD-L1 levels in patients with NPC compared to those in the controls. There was a significant difference in sPD-L1 levels between the controls and the patients with NPC (p = 0.031). p-value was calculated using the Mann–Whitney U-test. NPC: nasopharyngeal carcinoma, sPD-L1: soluble PD-L1.
Fig 3: Latent membrane protein 1 (LMP1) induces both cellular programmed death-ligand 1 (PD-L1) protein and soluble PD-L1 (sPD-L1). (Top lane) The media of both NP69T and NP69T-LMP1 cells were analyzed with western blotting for detecting PD-L1 levels. (Second, third, and fourth lane) Western blotting of PD-L1 and LMP1 in NP69T and NP69T-LMP1 cells. Total cell lysates and medium were collected from cell culture at 48 h. The harvested medium was concentrated to 50 fold.
Fig 4: Expression of PD-L1 (TPS and CPS) corelates with LMP1 in NPC tissue samples. Data were analyzed using the Spearman rank correlation coefficient. Statistical significance was defined as a p-value less than 0.05. (A) Relationship between LMP1 expression score and TPS (p = 0.023, r = 0.402). (B) Relationship between LMP1 expression score and CPS (p = 0.008, r = 0.461).
Fig 5: Kaplan–Meier curves of progression-free survival (PFS) in patients with NPC. The p-value was calculated using log-rank test. (A) Kaplan–Meier curves of LMP1-positive (=LMP1 expression score 10%) and LMP1-negative (PD-L1 TPS-positive (=10%) and PD-L1 TPS-negative (<10%) samples. (C) Kaplan–Meier curves of PD-L1 CPS-positive (=10%) and PD-L1 CPS-negative (<10%) samples. (D) Kaplan–Meier curves of sPD-L1-positive (=0.1 ng/mL) and sPD-L1-negative (<0.1 ng/mL) samples. * p < 0.05. LMP1: latent membrane protein 1, TPS: tumor proportion score, CPS: combined positive score, sPD-L1: soluble PD-L1.
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