Study on immune response induced by the fusion gene vaccine of gastric cancer MG7-Ag mimotope and heat shock protein 70 in mice
- VernacularTitle:胃癌MG7-Ag模拟表位与HSP70融合基因疫苗的构建及其诱导小鼠免疫反应的研究
- Author:
Xiaoxuan NING
;
Kaichun WU
;
Yua LI
- Publication Type:Journal Article
- Keywords:
stomach neoplasms;
vaccines, gene;
heat-shock proteins 70;
molecular mimicry
- From:
Medical Journal of Chinese People's Liberation Army
1982;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a fusion gene vaccine of gastric cancer MG7-Ag mimotope and heat shock protein 70, and to investigate the humoral and cellular immune response in mice induced by the vaccine. Methods The coding sequence of MG7 mimotope was incorporated with HSP70 gene at the 3' terminus by PCR amplification. Then the PCR product was cloned into pcDNA3.1(+) vector to construct the eukaryotic expression vector. The pcDNA3.1/MG7+hsp70 was sequenced to ensure the proper encoding. C57BL/6 mice were immunized with the fusion gene vaccine, and pcDNA3.1/hsp70 and empty pcDNA3.1 were used as controls. The serum was collected at the 3rd week after each immunization. Cellular ELISA was performed to evaluate the induced humoral immunity. The splenocytes were first separated at the 9th week for the assay of antigen specific CTL lysis activity by 51Cr release. Results A fragment about 2.0 kb was obtained by PCR amplification. Sequence analysis revealed that the sequence of mimotope was connected successfully to 3' terminus of hsp70 and the fusion gene was cloned into the pcDNA3.1(+) successfully. Cellular ELISA results suggested that the serum level of MG7-Ag antibody appeared in the vaccinated mice at 9th week, while no MG7-Ag antibody was detected in the controls. The results of the specific lysis rate of splenocytes showed no statistical difference between fusion gene vaccine group and control groups. Conclusion The fusion gene vaccine was constructed successfully and the specific humoral immune response was induced by the fusion gene vaccine.
