Surface expression of anti‐CD3scfv stimulates locoregional immunotherapy against hepatocellular carcinoma depending on the E1A‐engineered human umbilical cord mesenchymal stem cells
Surface expression of anti‐CD3scfv stimulates locoregional immunotherapy against hepatocellular carcinoma depending on the E1A‐engineered human umbilical cord mesenchymal stem cells
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DOI：
10.1002/ijc.30846
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发表时间：
2017-10
影响因子：
6.4
通讯作者：
Qing Zhang;Xiangfei Yuan;Yang Lu;Zhen-zhen Li;Shiqi Bao;Xiao-long Zhang;Yuanyuan Yang;D. Fan;Yi-zhi Zhang;Chenghan Wu;Hong-xing Guo;Yan-jun Zhang;Zhou Ye;D. Xiong
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文献类型：
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作者：
Qing Zhang;Xiangfei Yuan;Yang Lu;Zhen-zhen Li;Shiqi Bao;Xiao-long Zhang;Yuanyuan Yang;D. Fan;Yi-zhi Zhang;Chenghan Wu;Hong-xing Guo;Yan-jun Zhang;Zhou Ye;D. Xiong
肿瘤抗原是肿瘤免疫治疗的核心，然而，理想的抗原选择是困难的，特别是在免疫原性差的肿瘤中。在这项研究中，我们设计了一种策略，通过在肿瘤部位严格表达抗CD 3scFv来修饰肝细胞癌（HCC）细胞，这依赖于E1 A工程化的人脐带间充质干细胞（HUMSC.E1A）递送系统。随后，膜结合的抗CD 3scfv激活淋巴细胞，其绕过抗原表达或MHC限制裂解HCC细胞。首先，我们构建了由人甲胎蛋白（AFP）启动子驱动的抗CD 3scFv基因到腺病毒载体中，并将E1 A基因到慢病毒载体中。结果表明，抗CD 3单链抗体在体外能特异性表达于肝癌细胞表面，并激活淋巴细胞，有效杀伤靶细胞。AdCD 3scfv + LentiR.E1A感染HUMSC后36 h，HUMSC可支持腺病毒的复制和包装。使用皮下HepG 2异种移植模型，我们证实AdCD 3scfv和LentiR.E1A共转染的HUMSC可以选择性地迁移到肿瘤部位并产生大量的腺病毒。新产生的AdCD 3scfv成功感染并修饰肿瘤细胞。与对照组相比，MSC.E1A.AdCD3scfv和淋巴细胞注射组对肿瘤生长有明显的抑制作用。此外，5-氟尿嘧啶（5-FU）可以在低MOI下增敏腺病毒感染，从而提高体外和体内淋巴细胞的细胞毒性。综上所述，本研究为实体瘤免疫治疗提供了一种有前途的策略。
Tumor antigens is at the core of cancer immunotherapy, however, the ideal antigen selection is difficult especially in poorly immunogenic tumors. In this study, we designed a strategy to modify hepatocellular carcinoma (HCC) cells by surface expressing anti‐CD3scfv within the tumor site strictly, which depended on the E1A‐engineered human umbilical cord mesenchymal stem cells (HUMSC.E1A) delivery system. Subsequently, membrane‐bound anti‐CD3scfv actived the lymphocytes which lysed HCC cells bypassing the expression of antigens or MHC restriction. First, we constructed the anti‐CD3scfv gene driven by human α‐fetoprotein (AFP) promoter into an adenoviral vector and the E1A gene into the lentiviral vector. Our results showed that anti‐CD3scfv could specifically express on the surface of HCC cells and activate the lymphocytes to kill target cells effectively in vitro. HUMSC infected by AdCD3scfv followed by LentiR.E1A could support the adenoviral replication and packaging in vitro 36 h after LentiR.E1A infection. Using a subcutaneous HepG2 xenograft model, we confirmed that AdCD3scfv and LentiR.E1A co‐transfected HUMSC could migrate selectively to the tumor site and produce considerable adenoviruses. The new generated AdCD3scfv infected and modified tumor cells successfully. Mice injected with the MSC.E1A.AdCD3scfv and lymphocytes significantly inhibited the tumor growth compared with control groups. Furthermore, 5‐fluorouracil (5‐FU) could sensitize adenovirus infection at low MOI resulting in improved lymphocytes cytotoxicity in vitro and in vivo. In summary, this study provides a promising strategy for solid tumor immunotherapy.