制备人工抗原提呈细胞促进NK细胞增殖和抗肿瘤效应的研究  

The Promoting Effects of the Preparation of Artificial Antigen Presenting Cells in the Proliferation of NK Cells and the Research of Anti-tumor Effect

作　　者：吴凌娟 张磊 蔺广义 沈磊 WU Ling-juan;ZHANG Lei;LIN Guang-yi;SHEN Lei(Clinical Laboratory ofJianhai,Hebei Jianhai Biochip Corporation,Shijiazhuang 050035,China)

机构地区：[1]河北省健海生物芯片技术有限责任公司河北健海医学检验实验室,河北石家庄050035

出　　处：《标记免疫分析与临床》2018年第10期1556-1561,共6页Labeled Immunoassays and Clinical Medicine

基　　金：石家庄市科学技术研究与发展计划项目(编号:167200056A)

摘　　要：目的制备双表达CD86和4-1BBL的人工抗原提呈细胞,建立在体外大量的扩增具有较高杀伤活性的人NK细胞的有效方法。方法将表达CD86和4-1BBL的慢病毒感染K562细胞,制备成人工抗原提呈细胞(a APC),与人外周血单个核细胞(PBMC)共培养,使NK细胞在体外大量扩增。结果在培养d15天时,细胞数量达到2. 7×1010个,CD3-CD56+细胞纯度达到99. 4%,细胞体外杀伤活性为96. 27±1. 27%;在小鼠肺癌模型中,治疗组比对照组肿瘤体积明显减小。结论成功制备了有效扩增NK细胞的人工抗原提呈细胞(a APC),建立了在体外高效扩增具有较高杀伤活性的NK细胞的有效方法,为肿瘤和慢性感染性疾病的过继细胞免疫治疗奠定了基础。Objective To prepare artificial antigen- presenting cells expressing CD86 and 4- 1BBL,and establish an effective method for the proliferation of NK cells with high killing activity in vitro. Methods The lenti- virus expressing CD86- 4- 1- BBL was used to infect K562 cells,then prepared into the artificial antigen presenting cells （aAPC）.To increase the number of NK cells in vitro,the aAPC were co- cultured with peripheral blood mononuclear cells （PBMCs）. Results The cells number was 2.7×10 10 on the 15th day,with the purity of CD3- CD56＋cells reached 99.4%,and the cells activity in vitro was 96.27± 1.27% .In lung cancer models of mouse,the tumor volume reduced significantly in the treatment group compared with the control group. Conclusion Artificial antigen presenting cells （aAPC）expand NK cells in vitro is successfully achieved,and we establish an effective method for the proliferation of NK cells with high killing activity in vitro.It can serve as the foundation for the adoptive cellular immunotherapy of the cancer and chronic infectious diseases.

关 键 词：人工抗原提呈细胞(aAPC) CD86 4-1BBL NK细胞 高效扩增 杀伤活性 

分 类 号：R730.51[医药卫生—肿瘤]