表达抗PD-1抗体的溶瘤病毒在结肠癌腹腔移植瘤模型中的抗肿瘤作用  

Anti-Tumor Effects of Oncolytic Viruses Engineered with Anti-PD-1 Antibody in Celiac Graft Tumor Model of Colon Cancer

作　　者：刘文琦 胡宗风 李淑珍 李劲风 李小鹏 LIU Wenqi;HU Zongfeng;LI Shuzhen;LI Jinfeng;LI Xiaopeng(School of Pharmacy,Key Laboratory of Molecular Pharmacology and Drug Evaluation(Yantai University),Ministry of Education,Collaborative Innovation Center of Advanced Drug Delivery System and Biotech Drugs in Universities of Shandong,Yantai University,Yantai 264005,China)

机构地区：[1]烟台大学药学院,分子药理和药物评价教育部重点实验室(烟台大学),新型制剂与生物技术药物研究山东省高校协同创新中心,山东烟台264005

出　　处：《烟台大学学报（自然科学与工程版）》2024年第2期199-206,共8页Journal of Yantai University(Natural Science and Engineering Edition)

基　　金：山东省顶尖人才“一事一议”项目(主持人:李小鹏)。

摘　　要：为了研究表达小鼠抗PD-1抗体(VT1093M)的溶瘤病毒对结肠癌腹腔移植瘤模型的抗肿瘤作用,利用BALB/c小鼠成瘤的小鼠结肠癌CT26-luc细胞株建立了结肠癌腹腔移植瘤模型。接种后第8天进行分组,设置生理盐水(normal saline,NS)组、病毒VT09X组和VT1093M组,每组5只,注射剂量为2.5×10^(7)pfu/250μL,NS注射250μL,每隔一天注射一次,共注射5次。末次注射后6 d,脱颈处死小鼠,提取外周血中CD45^(+)细胞和腹水细胞,进行体外杀伤实验。治疗组初次出现荧光消失后第13天,进行再挑战实验。结果表明,造模实验确定最佳接种条件为200μL的2.5×10^(6)个/mL细胞悬液;药效实验中,VT09X组和VT1093M组较NS组具有明显的肿瘤抑制效果,VT1093M组治疗期间出现肿瘤消失情况,较VT09X组抑瘤效果明显,但差异无统计学意义;再挑战实验中,VT1093M组均未出现肿瘤再次生长;在体外杀伤实验中,注射病毒组的外周血CD45^(+)细胞在靶效比为1∶40时对CT26-luc杀伤效果明显,对4T1细胞,各组杀伤效果较差。因此,重组溶瘤病毒VT1093M在结肠癌腹腔转移瘤模型中具有显著的抗肿瘤作用,且能激发小鼠产生较为持久的特异性免疫反应。To investigate the anti-tumor effects of oncolytic viruses that express a mouse anti-PD-1 antibody(VT1093M)on a celiac graft tumor model of colon cancer,the peritoneal graft model using CT26-luc cells is established in BALB/c mice.On day 8 after inoculation,mice are randomly divided into three groups(n=5):normal saline(NS)group,VT09X virus group and VT1093M virus group.A dose of 2.5×10^(7)pfu/250μL is administered once every other day for a total of five viral injections.On day 6 after the last injection,mice are sacrificed,ascites cells and CD45^(+)cells from peripheral blood are extracted for in vitro killing experiments.The rechallenge experiment is performed on day 13 after the initial disappearance of fluorescence in the injected virus group.Modeling experiments determine that the optimal seeding condition is 200μL 2.5×10^(6)cells/mL cell suspension.The results indicate the potential efficacy of VT1093M oncolytic viruses in treating colon cancer.In the drug efficacy experiment,both the VT09X and VT1093M groups exhibit significant tumor suppression compared with the NS group.The VT1093M group shows complete tumor disappearance during the treatment period with a more pronounced tumor-suppressive effect than the VT09X group,though no significant difference is observed between the two groups.In the rechallenge experiment,no tumor regrowth is observed in the VT1093M group.In the in vitro killing experiment,the peripheral blood CD45^(+)cells in the injected virus group exhibit obvious killing effect on CT26-luc when the target effect ratio is 1∶40,while the killing effect on 4T1 cells is poor.These findings demonstrate that the recombinant oncolytic virus VT1093M has a significant antitumor effect in the colon cancer abdominal metastasis model and it stimulates a relatively durable specific immune response in mice.

关 键 词：结肠癌 腹腔移植瘤 溶瘤病毒 生物发光 特异性免疫 

分 类 号：R735.35[医药卫生—肿瘤]