共表达IL-7和CCL19的VSVΔ51溶瘤病毒在肝癌中的抗肿瘤活性研究  

Coexpressing IL-7 and CCL19 antitumor activity of VSVΔ51 oncolyticvirus in hepatocellular carcinoma

作　　者：宋娟丽 史娜[2] 王志芬[1] 冀青青 SONG Juanli;SHI Na;WANG Zhifen;JI Qingqing(The Third Department of Oncology,Handan Central Hospital,Handan 056000,China;The Fourth Department of Liver,Handan Infectious Diseases Hospital,Handan 056000,China)

机构地区：[1]河北省邯郸市中心医院肿瘤三科,河北邯郸056000 [2]河北省邯郸市传染病医院肝四科,河北邯郸056000

出　　处：《东南大学学报（医学版）》2023年第4期526-532,共7页Journal of Southeast University(Medical Science Edition)

基　　金：河北省医学科学研究课题(20220013)。

摘　　要：目的:构建共表达白细胞介素7(interleukin-7,IL-7)和趋化因子配体19[chemokine(C-C motif) ligand 19,CCL19]的溶瘤病毒VSVΔ51(7×19 VSVΔ51),并探索其在肝癌中的抗肿瘤活性。方法:细胞的活力测定(MTS法)检测溶瘤病毒对肿瘤细胞的细胞毒性;ELISA法检测病毒感染上清及肿瘤组织匀浆中的IL-7和CCL19的含量;皮下移植瘤模型检测溶瘤病毒的肿瘤抑制能力;流式细胞术检测肿瘤浸润T细胞的表型;免疫组化法检测肿瘤浸润T细胞及IFN-γ的含量。结果:VSVΔ51和7×19 VSVΔ51病毒均可以在小鼠肝癌细胞H22和Hepa1-6中复制,在2 PFU时裂解细胞的效率接近100%;VSVΔ51和7×19 VSVΔ51病毒均对小鼠肝癌移植瘤的生长产生抑制作用,且7×19 VSVΔ51病毒较VSVΔ51病毒抑制能力更强[瘤质量(232.7±13.72) mg vs.(603.8±20.3) mg,P<0.000 1];7×19 VSVΔ51病毒组治疗后肿瘤组织中IL-7[(673.5±62.6)pg·ml^(-1)vs.(383.1±108.3) pg·ml^(-1),P<0.01]和CCL19[(756.2±41.6)pg·ml^(-1)vs.(356.2±50.2) pg·ml^(-1),P<0.001]的含量较VSVΔ51病毒组显著升高,肿瘤浸润T细胞的数目增多(805±133 vs. 392±21,P<0.05),干细胞样记忆T细胞的比例显著提升[(72.5±9.8)%vs.(49.3±8.5)%,P<0.001)],肿瘤组织中IFN-γ的含量也明显升高[(817±52) pg·ml^(-1)vs.(227±63) pg·ml^(-1),P<0.001)]。结论:7×19 VSVΔ51病毒可在肝癌小鼠模型中激发出强大的抗肿瘤免疫反应并发挥显著的抗肿瘤作用。Objective:To construct an oncolytic virus VSVΔ51 overexpressing IL-7 and CCL19(7×19 VSVΔ51),and to explore its antitumor activity in hepatocellular carcinoma.Methods:MTS assay was used to detect the cytotoxicity of oncolytic virus on tumor cells.The contents of IL-7 and CCL19 in cell supernatant after viral infection and tumor tissue homogenate were detected by ELISA.Subcutaneous tumor transplantation model were used to verify the tumor suppressive ability of oncolytic virus.The phenotype of tumor infiltrating T cells was detected by flow cytometry.The contents of T cells and IFN-γwere detected by immunohistochemistry.Results:Both VSVΔ51 and 7×19 VSVΔ51 could replicate and lyse tumor cells in mouse hepatoma cells H22 and Hepa1-6.Both VSVΔ51 and 7×19 VSVΔ51 could inhibit the growth of hepatocellular carcinoma xenografts in mice,but 7×19 VSVΔ51 had a stronger inhibitory effect than VSVΔ51[tumor weight(232.7±13.72)mg vs.(603.8±20.3)mg,P<0.0001].After 7×19 VSVΔ51 treatment,the contents of IL-7[(673.5±62.6)pg·ml^(-1) vs.(383.1±108.3)pg·ml^(-1),P<0.01]and CCL19[(756.2±41.6)pg·ml^(-1) vs.(356.2±50.2)pg·ml^(-1),P<0.001]in tumor tissues were significantly higher than those in VSVΔ51 group,the number of tumors infiltrating T cells was increased(805±133 vs.392±21,P<0.05),the proportion of stem cell-like memory T cells was significantly increased[(72.5±9.8)%vs.(49.3±8.5)%,P<0.001)],and the content of IFN-γin tumor tissues was also significantly increased[(817±52)pg·ml^(-1) vs.(227±63)pg·ml^(-1),P<0.001)].Conclusion:7×19 VSVΔ51 could stimulate a strong antitumor immune response and play a significant antitumor effect in the hepatocellular carcinoma xenografts model.

关 键 词：肝癌 溶瘤病毒 水泡性口炎病毒Δ51 白细胞介素7 趋化因子配体19 小鼠 

分 类 号：R735.7[医药卫生—肿瘤]