重庆医学2017，Vol.46Issue(19)：2612-2614,3.DOI:10.3969/j.issn.1671-8348.2017.19.006
基于SOE-PCR的核酸疫苗pIRES2-MLAA34-HSP70的构建及免疫效果研究
Study on immune effect of DNA vaccine pIRES2-MLAA34-HSP70 constructed by SOE-PCR
摘要
Abstract
Objective To construct the DNA vaccine pIRES2-MLAA34-HSP70,and to detect its immune effect.Methods The acute monocytic leukemia associated antigen gene MLAA-34 and heat-shock protein (HSP)70 gene were extracted by using RT-PCR.The specific overlapping primer was designed,and the fusion gene MLAA34-HSP70 was amplified by using SOE-PCR technique.Then the DNA vaccine pIRES2-MLAA34-HSP70 was constructed,and BALB/c mice were immunized with this DNA vaccine.The splenic lymphocyte killing activity was detected by using MTT,levels of IL-2,IL-4 and IFN-γ were also detected by using ELISA.Results The MLAA34-HSP70 gene (2 956 bp) and the DNA vaccine pIRES2-MLAA34-HSP70 was amplified and constructed successfully.The killing efficiency of DNA vaccine pIRES2-MLAA34-HSP70 in U937 cells was significantly higher than that in other experimental groups and control group (P＜0.01),and levels of IL-4,IL-2 and IFN-γin DNA vaccine pIRES2-MLAA34-HSP70 group were significantly higher than those in the other experimental groups and control group (P＜0.01).Conclnsion The DNA vaccine pIRES2-MLAA34-HSP70 is constructed successfully.It is shown that the DNA vaccine induces strong humoral immunity,which could enhance immune responses to tumor cells and specificlly kill MLAA34 positive cells.关键词
疫苗,DNA/重叠延伸PCR/白血病,单核细胞,急性/MLAA-34/热休克蛋白质类70Key words
vaccines,DNA/gene splicing by overlap extension PCR/leukemia,monocytic,acute/MLAA-34/heat-shock protein 70分类
医药卫生引用本文复制引用
赵佳琪,王婉莹,范业宁,马春平,张东红,吕扬,赵臣..基于SOE-PCR的核酸疫苗pIRES2-MLAA34-HSP70的构建及免疫效果研究[J].重庆医学,2017,46(19):2612-2614,3.基金项目
吉林省科技厅自然科学基金资助项目(20160101179JC) （20160101179JC）
吉林省卫生和计划生育委员会项目(2015Z071) （2015Z071）
吉林省教育厅“十三五”科技项目(JJKH20170416KJ) （JJKH20170416KJ）
吉林省大学生创新创业课题(2015024) （2015024）
国家级大学生创新创业训练计划项目(201613706019). （201613706019）
