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基于胃癌MG(7)-Ag模拟表位的口服DNA疫苗的研制

[Development of oral DNA vaccine based on MG(7)-Ag mimotope of gastric cancer].

作者信息

Guo Changcun, Ding Jie, Yu Zhaocai, Han Quanli, Meng Fanping, Liu Na, Fan Daiming

机构信息

PLA Institute of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2002 Mar;24(2):110-3.

Abstract

OBJECTIVE

To develop an oral DNA vaccine based on MG(7)-Ag mimotope of gastric cancer using attenuated Salmonella typhimurium and evaluate its efficacy and protective effect.

METHODS

The eukaryotic expression vector including the MG(7)-Ag mimotope and a Th epitope was constructed, and then transduced into an attenuated Salmonella typhimurium to get the oral DNA vaccine. C57BL/6 J mice were orally immunized with 1 x 10(8) cfu Salmonella transfectants, with Salmonella harboring empty plasmid, with phophate buffered saline (PBS) as control. At the 6th week, serum titer of MG(7) antibody was detected by ELISA. In the 8th week, a [(3)H]-thymidine incorporation assay was performed to test the proliferation of murine spleen cells to the stimulant of MG(7)-Ag mimicry peptide. At the same time, Ehrlich ascites carcinoma cells expressing MG(7)-Ag were used in tumor challenge assay to evaluate the protective effect of the immunization.

RESULTS

The oral DNA vaccine induced MG(7) antibody in mice, while in vivo unprimed proliferation assay of the spleenocytes showed no difference among the three groups. Two weeks after tumor challenge, 2 in 7 immunized mice were tumor free, while none in the control group was protected.

CONCLUSION

Oral DNA vaccine based on the MG(7)-Ag momitope is immunogenic. It is able to induce specific immunity response against tumor in mice, and the vaccine is partially protective.

摘要

目的

利用减毒鼠伤寒沙门菌研制基于胃癌MG(7)-Ag模拟表位的口服DNA疫苗,并评价其疗效和保护作用。

方法

构建含MG(7)-Ag模拟表位和Th表位的真核表达载体,然后转导入减毒鼠伤寒沙门菌获得口服DNA疫苗。将C57BL/6 J小鼠分别用1×10(8) cfu转染的沙门菌、携带空质粒的沙门菌、磷酸盐缓冲液(PBS)口服免疫作为对照。第6周,用ELISA法检测MG(7)抗体血清滴度。第8周,进行[(3)H]-胸腺嘧啶核苷掺入试验,检测小鼠脾细胞对MG(7)-Ag模拟肽刺激物的增殖情况。同时,用表达MG(7)-Ag的艾氏腹水癌细胞进行肿瘤攻击试验,评价免疫的保护作用。

结果

口服DNA疫苗可诱导小鼠产生MG(7)抗体,而脾细胞体内未致敏增殖试验在三组间无差异。肿瘤攻击2周后,7只免疫小鼠中有2只无肿瘤,而对照组无一只得到保护。

结论

基于MG(7)-Ag模拟表位的口服DNA疫苗具有免疫原性。它能诱导小鼠产生针对肿瘤的特异性免疫反应,且该疫苗具有部分保护作用。

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