减毒沙门氏菌VNP20009的抗肿瘤机制研究  

Research on anti-tumor mechanism of attenuated Salmonella typhimurium VNP20009

作　　者：尹特 刘丽娜 董世达 黄宝连 李晨阳 曹志婷 华子春 YIN Te;LIU Li-na;DONG Shi-da;HUANG Bao-lian;LI Chen-yang;CAO Zhi-ting;HUA Zi-chun(School of Biopharmacy,China Pharmaceutical University,Nanjing 211198,China;The State Key Laboratory of Pharmaceutical Biotechnology,College of Life Sciences,Nanjing University,Nanjing 210033,China;Institute of Pharmaceutical Biotechnology of Jiangsu Industrial Technology Research Institute,Changzhou 213164,China)

机构地区：[1]中国药科大学生物药物学院,江苏南京211198 [2]南京大学生命科学学院,医药生物技术国家重点实验室,江苏南京210033 [3]江苏省产业技术研究院医药生物技术研究所,江苏常州213164

出　　处：《药学学报》2023年第9期2700-2706,共7页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(32250016,82130106);南京市生命健康科技专项计划(202110016);常州市科技局(CZ20210010,CJ20210024,CJ20220019)。

摘　　要：减毒沙门氏菌VNP20009是一种应用范围较广的天然来源溶瘤细菌,基于其已证实的临床安全性、可特异性趋化靶向肿瘤和明确已知的基因组序列等优点而被广泛用于抗肿瘤研究。本研究建立黑色素瘤小鼠模型(所有动物实验均遵循中国药科大学动物伦理委员会的规定),通过腹腔注射VNP20009对其抗肿瘤活性进行验证,与对照组相比,VNP20009治疗可显著抑制肿瘤生长;体外共培养实验证明VNP20009能够诱导巨噬细胞向M1表型极化并表达相关炎症因子;通过流式细胞实验分析肿瘤和肿瘤引流淋巴结(tumor-draining lymph node,TDLN)内免疫变化,证明VNP20009治疗诱导肿瘤组织免疫细胞增加,进一步分析发现肿瘤引流淋巴结内T细胞浸润增加,而VNP20009治疗能够诱导肿瘤内部的CD4^(+)T和CD8^(+)T细胞活化。本研究结果证明,VNP20009可通过诱导巨噬细胞向M1表型极化,招募并激活细胞毒性T细胞,协同其自身组分,共同抑制小鼠体内黑色素瘤的生长。Attenuated Salmonella typhimurium VNP20009 is a widely used natural oncolytic bacterium,which has great application potential given its unique characteristics,including clinical safety,tumor targeting specificity,and explicit genome sequence.Here,we show that tumor progression can be effectively reduced by intraperitoneal administration with VNP20009 in a mouse model of melanoma(all animal experiments were conducted in accordance with the Animal Ethics Committee of China Pharmaceutical University);co-culture experiment in vitro demonstrated that VNP20009 can induce the polarization of macrophage M1,accompanied by expression of inflammation-related factors;flow cytometry analysis showed that VNP20009 induced the increase of immune cell infiltration in tumor.Further analysis showed that T cells infiltration in tumor-draining lymph node(TDLN)increased,and VNP20009 induced the activation of CD4^(+)T cells and CD8^(+)T cells in tumor.Our results demonstrate that VNP20009 treatment significantly inhibited melanoma tumors by remodeling tumor-associated macrophages to an M1-like phenotype,as well as recruiting and activating cytotoxic T cells,combined with its own antigenic activity to exert anti-tumor immunity.

关 键 词：VNP20009 黑色素瘤 巨噬细胞 肿瘤引流淋巴结 细胞毒性T细胞 

分 类 号：R966[医药卫生—药理学]