负载胃癌新抗原肽和CD73抑制剂纳米粒子的中性粒细胞疫苗的制备和抗肿瘤效果评价  

Preparation and evaluation of neutrophil-loaded neoantigen peptide and CD73 inhibitor nanoparticles vaccine on gastric cancer

作　　者：翟青青 褚雁鸿 秦岚群 张小力 周熙鹏 刘芹[2] 刘宝瑞[1] ZHAI Qingqing;CHU Yanhong;QIN Lanqun;ZHANG Xiaoli;ZHOU Xipeng;LIU Qin;LIU Baorui(The Comprehensive Cancer Centre,Nanjing Drum Tower Hospital,Clinical College of Nanjing Medical University,Jiangsu Nanjing 210008,China;the Comprehensive Cancer Centre,Nanjing Drum Tower Hospital,the Affiliated Hospital of Nanjing University Medical School,Jiangsu Nanjing 210008,China)

机构地区：[1]南京医科大学鼓楼临床医学院肿瘤中心,江苏南京210008 [2]南京大学医学院附属鼓楼医院肿瘤中心,江苏南京210008

出　　处：《现代肿瘤医学》2023年第1期24-32,共9页Journal of Modern Oncology

基　　金：江苏省卫健委面上项目(编号:M2020035);江苏省南京市卫健委重点项目(编号:ZKX19012)。

摘　　要：目的:制备负载胃癌细胞新抗原肽和CD73抑制剂的中性粒细胞(NEs)疫苗并表征,初步评价其在荷瘤615小鼠中的抗肿瘤效果。方法:通过Percoll梯度密度离心法分离纯化小鼠骨髓来源的NEs。分别制备仅负载胃癌细胞来源新抗原肽疫苗(DP-Ag-MFC-NEs),以及同时负载新抗原肽和CD73抑制剂纳米粒子的联合疫苗(DP-Ag-MFC-NEs+antiCD73+CpG),观察疫苗对荷瘤小鼠的治疗效果。结果:一只10周龄615小鼠可分离纯化到平均约1.5×10^(6)个NEs,细胞活性在95%以上,纯度在90%以上。和对照组相比,DP-Ag-MFC-NEs可显著抑制小鼠皮下肿瘤生长,使部分小鼠痊愈(CR),小鼠淋巴结中CD4^(+)、CD8^(+)阳性细胞比例显著增高(P<0.05)。CR小鼠再次接种MFC胃癌细胞,没有小鼠肿瘤复发。联合疫苗的治疗效果确切,小鼠存活率最高(75%,P=0.038)。结论:负载胃癌细胞来源的新抗原肽和CD73抑制剂的中性粒细胞疫苗对小鼠无明显毒性,可显著抑制肿瘤生长,甚至治愈小鼠。同时,该疫苗可在小鼠体内激活记忆型免疫应答。Objective:To prepare and characterize neutrophils(NEs)-loaded neoantigen peptide and CD73 inhibitor nanoparticles(NPs)vaccine,and to preliminarily evaluate its antitumor effect in gastric tumor-bearing 615 mice.Methods:NEs mouse bone marrow derived NEs were isolated and purified by Percoll gradient density centrifugation.The gastric cancer cells MFC derived neoantigen peptide were linked with DSPE-PEG_(2000)(DP-MFC)and the CD73 inhibitor NPs(antiCD73)were prepared by phacoemulsification and solvent evaporation technology.The vaccine was prepared by co-incubating the NEs with DP-MFC and adjuvant(DP-MFC-NEs vaccine)or both DP-MFC and antiCD73NPs with adjuvant(combined vaccine).The therapeutic effect of the vaccines on gastric cancer was observed in tumor-bearing 615 mice.Results:About 1.5×10^(6)NEs could be isolated and purified from a10-week-old 615 mouse.The cell activity and purity were above 95%and 90%respectively.Compared with control mice,the growth of subcutaneous tumor in mice treated with DP-MFC-NEs vaccine was significantly inhibited,and some of them were cured(CR).The proportions of CD4^(+)and CD8^(+)cells in their lymph nodes were significantly higher(P<0.05).Those CR mice were subcutaneously inoculated with MFC cells again,and no tumor recurred.Mice treated with the combined vaccine had the highest survival rate(75%,P=0.038).Conclusion:Neutrophil vaccine loaded with neoantigen peptides derived from gastric cancer cells and CD73 inhibitor NPs has no obvious toxicity to mice,and can inhibit tumor growth and even cure mice.The vaccine could activate a memory-type immune response in mice.

关 键 词：胃癌 MFC细胞 新抗原肽 中性粒细胞 疫苗 CD73抑制剂 纳米粒子 

分 类 号：R735.2[医药卫生—肿瘤]