Study on immune effect of DNA vaccine pIRES2-MLAA34-HSP70 constructed by SOE-PCR
10.3969/j.issn.1671-8348.2017.19.006
- VernacularTitle:基于SOE-PCR的核酸疫苗pIRES2-MLAA34-HSP70的构建及免疫效果研究
- Author:
Jiaqi ZHAO
;
Wanying WANG
;
Yening FAN
;
Chunping MA
;
Donghong ZHANG
;
Yang LV
;
Chen ZHAO
- Keywords:
vaccines,DNA;
gene splicing by overlap extension PCR;
leukemia,monocytic,acute;
MLAA-34;
heat-shock protein 70
- From:
Chongqing Medicine
2017;46(19):2612-2614
- CountryChina
- Language:Chinese
-
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.
