Faculty of Hepato-Pancreato-Biliary Surgery, the First Medical Centre, Chinese PLA General Hospital, Institute of Hepatobiliary Surgery of Chinese PLA, Key Laboratory of Digital Hepatobiliary Surgery, PLA, Beijing, China; National Clinical Research Center for Infectious Diseases, the Fifth Medical Center of PLA General Hospital, Beijing, China.
Faculty of Hepato-Pancreato-Biliary Surgery, the First Medical Centre, Chinese PLA General Hospital, Institute of Hepatobiliary Surgery of Chinese PLA, Key Laboratory of Digital Hepatobiliary Surgery, PLA, Beijing, China.
Int Immunopharmacol. 2023 Jul;120:110323. doi: 10.1016/j.intimp.2023.110323. Epub 2023 May 17.
To evaluate targeted killing of hepatocellular carcinoma (HCC) cells by a recombinant oncolytic influenza virus expressing a PD-L1 antibody (rgFlu/PD-L1) and to develop a novel immunotherapy for HCC.
Using influenza virus reverse genetics, a recombinant oncolytic virus was generated in the background of the A/Puerto Rico/8/34 (PR8) virus, then identified via screening and passage in specific pathogen-free chicken embryos. Hepatocellular carcinoma cell killing by rgFlu/PD-L1 was confirmed in vitro and in vivo. Transcriptome analyses were used to explore PD-L1 expression and function. Western blotting revealed that PD-L1 activated the cGas-STING pathway.
rgFlu/PD-L1 expressed the PD-L1 heavy and light chain in PB1 and PA, respectively; PR8 served as the backbone. The hemagglutinin titer of rgFlu/PD-L1 was 2, and the virus titer was 9-10 logTCID/mL. Electron microscopy revealed that the rgFlu/PD-L1 morphology and size were consistent with wild-type influenza virus. The MTS assay showed that rgFlu/PD-L1 induced significant killing of HCC cells but not normal cells. rgFlu/PD-L1 inhibited PD-L1 expression and induced apoptosis in HepG2 cells. Notably, rgFlu/PD-L1 controlled the viability and function of CD8 T cells by activating the cGas-STING pathway.
rgFlu/PD-L1 activated the cGas-STING pathway in CD8 T cells, causing them to kill HCC cells. This approach represents a novel immunotherapy for liver cancer.
评估表达 PD-L1 抗体的重组溶瘤流感病毒（rgFlu/PD-L1）对肝癌细胞的靶向杀伤作用，并为肝癌开发一种新的免疫疗法。
利用流感病毒反向遗传学技术，在 A/Puerto Rico/8/34（PR8）病毒背景下生成重组溶瘤病毒，然后通过在无特定病原体鸡胚中的筛选和传代进行鉴定。体外和体内实验证实 rgFlu/PD-L1 对肝癌细胞的杀伤作用。通过转录组分析探索 PD-L1 的表达和功能。Western blot 显示 PD-L1 激活了 cGas-STING 通路。
rgFlu/PD-L1 在 PB1 和 PA 中分别表达 PD-L1 重链和轻链，PR8 作为骨架。rgFlu/PD-L1 的血凝素效价为 2，病毒效价为 9-10 logTCID/mL。电子显微镜显示 rgFlu/PD-L1 的形态和大小与野生型流感病毒一致。MTS 检测表明 rgFlu/PD-L1 诱导肝癌细胞明显杀伤，但对正常细胞无影响。rgFlu/PD-L1 抑制 HepG2 细胞中 PD-L1 的表达并诱导其凋亡。值得注意的是，rgFlu/PD-L1 通过激活 cGas-STING 通路控制 CD8 T 细胞的活力和功能。
rgFlu/PD-L1 通过激活 CD8 T 细胞中的 cGas-STING 通路，导致其杀伤肝癌细胞。这种方法为肝癌提供了一种新的免疫治疗策略。
