Meng Fan-Ping, Ding Jie, Yu Zhao-Cai, Han Quan-Li, Guo Chang-Cun, Liu Na, Fan Dai-Ming
Institute of Digestive Disease, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, Shaanxi Province, China.
World J Gastroenterol. 2005 Mar 28;11(12):1833-6. doi: 10.3748/wjg.v11.i12.1833.
To develop an oral attenuated Salmonella typhimurium vaccine against gastric cancer and to evaluate its efficacy in mice.
A complementary sequence of Nco I site and a sequence coding for MG7-Ag mimotope were designed at the 5' terminus of forward primer. Using p1.2 II-HBCAg plasmid as template, PCR was performed to get a fusion gene of the mimotope and a HBcAg gene. The fusion gene was then subcloned into the plasmid pYA3341 complementary to Salmonella typhimurium X4550, and the recombinant plasmid was then transformed into attenuated Salmonella typhimurium X4550. Balb/c mice were orally immunized with the recombinant Salmonella typhimurium X4550. The mice were immunized every 2 wk to reinforce the immunity. At the 6th wk, serum titer of antibody was detected by ELISA, and at the 8th wk, cellular immunity was detected by (51)Cr release test. Ehrlich ascites carcinoma cells expressing MG7-Ag were used in tumor challenge assay as a model to evaluate the protective effect of the vaccine.
Serum titer of antibody against MG7-Ag was significantly higher in mice immunized with the vaccine than in control groups (0.9538+/-0.043 vs 0.6531+/-0.018, P<0.01; 0.9538+/-0.043 vs 0.6915+/-0.012, P<0.01), while in vitro (51)Cr release assay of the splenocytes showed no statistical difference in the three groups. Two weeks after tumor challenge, 1 in 5 immunized mice was tumor free, while all the mice in the control group presented tumor.
Oral attenuated Salmonella typhimurium vaccine against the MG7-Ag mimotope of gastric cancer is immunogenic. It can induce significant humoral immunity against tumors in mice, and has some protective effects.
研制一种针对胃癌的口服减毒鼠伤寒沙门氏菌疫苗,并评估其在小鼠体内的疗效。
在正向引物的5'末端设计Nco I位点的互补序列和编码MG7-Ag模拟表位的序列。以p1.2 II-HBCAg质粒为模板,进行PCR以获得模拟表位与乙肝核心抗原(HBcAg)基因的融合基因。然后将融合基因亚克隆到与鼠伤寒沙门氏菌X4550互补的质粒pYA3341中,再将重组质粒转化到减毒鼠伤寒沙门氏菌X4550中。用重组鼠伤寒沙门氏菌X4550对Balb/c小鼠进行口服免疫。每2周免疫小鼠一次以加强免疫。在第6周时,通过酶联免疫吸附测定(ELISA)检测血清抗体滴度,在第8周时,通过铬(51)释放试验检测细胞免疫。使用表达MG7-Ag的艾氏腹水癌细胞作为肿瘤攻击试验模型来评估疫苗的保护作用。
用疫苗免疫的小鼠中,抗MG7-Ag的血清抗体滴度显著高于对照组(0.9538±0.043对0.6531±0.018,P<0.01;0.9538±0.043对0.6915±0.012,P<0.01),而脾细胞的体外铬(51)释放试验在三组中无统计学差异。肿瘤攻击两周后,5只免疫小鼠中有1只无肿瘤,而对照组所有小鼠均出现肿瘤。
针对胃癌MG7-Ag模拟表位的口服减毒鼠伤寒沙门氏菌疫苗具有免疫原性。它能在小鼠体内诱导显著的抗肿瘤体液免疫,并具有一定的保护作用。