PD-1 ScFv溶瘤病毒对卵巢癌裸鼠模型免疫活性、肿瘤生长及存活率的影响  

Impacts of PD-1 ScFv oncolytic virus on immune activity,tumor growth and survival in nude mouse model of ovarian cancer

作　　者：黄山鹰[1] 唐国玲[1] 胡海燕[1] 刘剀[1] 罗智华[1] 曾薇薇[1] 帅振虹 刘植华[1] HUANG Shanying;TANG Guoling;HU Haiyan;LIU Kai;LUO Zhihua;ZENG Weiwei;SHUAI Zhenhong;LIU Zhihua(Department of Gynaecology,Shenzhen Maternal and Child Health Hospital Affiliated to Southern Medical University,Shenzhen 518000,China)

机构地区：[1]深圳市妇幼保健院妇科,518000

出　　处：《免疫学杂志》2022年第10期854-861,共8页Immunological Journal

基　　金：广东省医学科学技术研究基金(B2021289)。

摘　　要：目的探讨程序性细胞死亡蛋白1(PD-1)单链抗体(ScFv)溶瘤病毒对卵巢癌裸鼠模型免疫活性、肿瘤生长及存活率的影响。方法C57BL/6小鼠右前腋下注射SKOV-3细胞构建卵巢癌裸鼠模型,分别以0、5×10^(5)PFU、1×10^(6)PFU、5×10^(6)PFU、1×10^(7)PFU、5×10^(7)PFU、1×10^(8)PFU滴度的PD-1 ScFv溶瘤病毒瘤内注射,检测小鼠肿瘤体积,筛选最佳作用滴度。构建好的卵巢癌模型裸鼠随机分为模型组、PD-1抗体组、单独溶瘤病毒组、PD-1 ScFv溶瘤病毒组,每组8只,再选8只小鼠右前腋下注射等量细胞培养液作为对照组。测定小鼠脾指数、胸腺指数及肿瘤体积;流式细胞实验检测小鼠外周血T淋巴细胞亚群CD4^(+)、CD8^(+)比例与CD4^(+)/CD8^(+)比值;以酶联免疫吸附法(ELISA)试剂盒检测小鼠血清白细胞介素(IL)-2、IL-10、γ-干扰素(IFN-γ)水平;检测小鼠外周血白细胞比例;CCK-8实验检测小鼠产生的特异性抗肿瘤细胞杀伤率;检测各组小鼠存活率。结果与对照组比较,模型组小鼠脾指数与胸腺指数、CD4^(+)比例与CD4^(+)/CD8^(+)比值、IFN-γ与IL-2水平、外周血白细胞比例、特异性抗肿瘤细胞杀伤率、存活率降低(P<0.05)、CD8^(+)比例、IL-10水平升高(P<0.05);与模型组比较,PD-1 ScFv溶瘤病毒组、单独溶瘤病毒组、PD-1抗体组小鼠脾指数与胸腺指数、CD4^(+)比例与CD4^(+)/CD8^(+)比值、IFN-γ与IL-2水平、外周血白细胞比例、特异性抗肿瘤细胞杀伤率、存活率均升高(P<0.05),肿瘤体积、CD8^(+)比例、IL-10水平均降低(P<0.05);与单独溶瘤病毒组、PD-1抗体组分别比较,PD-1 ScFv溶瘤病毒组小鼠脾指数与胸腺指数、CD4^(+)比例与CD4^(+)/CD8^(+)比值、IFN-γ与IL-2水平、外周血白细胞比例、特异性抗肿瘤细胞杀伤率、存活率均升高(P<0.05),肿瘤体积、CD8^(+)比例、IL-10水平均降低(P<0.05)。结论PD-1 ScFv溶瘤病毒可改善卵巢癌裸鼠免疫功能,促进特异性抗肿瘤细胞产�This study was designed to investigate the impacts of programmed cell death protein 1(PD-1)single chain antibody(ScFv)oncolytic virus on the immune activity,tumor growth and survival rate of ovarian cancer nude mouse model.SKOV-3 cells were injected into the right anterior axilla of C57BL/6 mice to construct a nude mouse model of ovarian cancer,then the PD-1 ScFv oncolytic virus was injected intratumorally with titers of 0,5×10^(5) PFU,1×10^(6) PFU,5×10^(6) PFU,1×10^(7) PFU,5×10^(7) PFU,and 1×10^(8) PFU,respectively.The volume of tumor in the mice was measured,and the optimal effect titer was screened.Nude mouse models of ovarian cancer were randomly separated into model group,PD-1 antibody group,oncolytic virus alone group,and PD-1 ScFv oncolytic virus group,with 8 mice in each group,and control group was set up in 8 normal mice by injecting the same amount of cell culture solution in the right anterior axilla.Mouse spleen index,thymus index and tumor volume were determined;flow cytometry was performed to measure the ratio of CD4^(+),CD8^(+)and CD4^(+)/CD8^(+)in peripheral blood;ELISA kits were used to measure the levels of serum interleukin(IL)-2,IL-10 andγ-interferon(IFN-γ);the proportion of leukocytes in peripheral blood of mice was detected;CCK-8 assay was performed to measure the specific anti-tumor cell killing rate of mice;the survival rate of mice in each group was measured.Data showed that the spleen index and thymus index,CD4^(+)proportion and CD4^(+)/CD8^(+)ratio,IFN-γand IL-2 levels,proportion of peripheral blood leukocytes,specific anti-tumor cell killing rate and survival rate of mice in the model group decreased as compared with control group(P<0.05),while CD8^(+)ratio and IL-10 level increased(P<0.05);as compared with model group,all these indexes mentioned above were reversed,and the tumor volume was diminished in PD-1 ScFv oncolytic virus group,oncolytic virus alone group and PD-1 antibody group(P<0.05),especially in the PD-1 ScFv oncolytic virus group(P<0.05).In conclusion,PD-1 ScFv o

关 键 词：程序性死亡受体1单链抗体 溶瘤病毒 卵巢癌 裸鼠模型 免疫活性 

分 类 号：R737.31[医药卫生—肿瘤]