Todoroki I, Joh T, Watanabe K, Miyashita M, Seno K, Nomura T, Ohara H, Yokoyama Y, Tochikubo K, Itoh M
First Department of Internal Medicine, Department of Microbiology, Nagoya City University Medical School, 1 Kawasumi, Mizuho-Cho, Mizuho-Ku, Nagoya, 467-8601, Japan.
Biochem Biophys Res Commun. 2000 Oct 14;277(1):159-63. doi: 10.1006/bbrc.2000.3632.
We investigated the effect of DNA vaccines encoding H. pylori-heat shock protein A and B (pcDNA3.1-hspA and -hspB) on inducing immune responses against H. pylori in mice. C57BL/six mice aged 5 weeks were immunized by single injection of 10 microg of pcDNA3.1-hspA and pcDNA3.1-hspB into intracutaneous tissue. Plasmid DNA lacking the inserted hsp were injected as a control. Three months after vaccination, significant specific antibodies against H. pylori were detected by ELISA in the sera of vaccinated mice. Antibody isotypes were predominantly IgG2a (Th1-like) with pcDNA3.1-hspA and mixed IgG1/IgG2a (Th0-like) with pcDNA3.1-hspB. DNA vaccination dramatically suppressed colonies of bacteria in stomach of vaccinated mice (28,400 +/- 21,600/mm(2) for pcDNA3.1-hspA and 6800 +/- 3470/mm(2) for pcDNA3.1-hspB) compared to control mice (128,000 +/- 42,200/mm(2)). Histological analysis of the gastric mucosa demonstrated that the degree of gastritis was significantly lower in the vaccinated mice than in control mice. These results demonstrated that DNA vaccines encoding H. pylori-Hsp induce significant immune response against H. pylori to decrease gastric mucosal inflammation, indicating that a DNA vaccine can be a new approach against H. pylori in humans.
我们研究了编码幽门螺杆菌热休克蛋白A和B的DNA疫苗(pcDNA3.1-hspA和-hspB)对诱导小鼠抗幽门螺杆菌免疫反应的影响。将5周龄的C57BL/6小鼠皮内单次注射10微克pcDNA3.1-hspA和pcDNA3.1-hspB进行免疫。注射缺乏插入hsp的质粒DNA作为对照。接种疫苗三个月后,通过ELISA在接种疫苗小鼠的血清中检测到针对幽门螺杆菌的显著特异性抗体。抗体亚型以pcDNA3.1-hspA为主的IgG2a(Th1样)和以pcDNA3.1-hspB为主的混合IgG1/IgG2a(Th0样)。与对照小鼠(128,000±42,200/mm²)相比,DNA疫苗显著抑制了接种疫苗小鼠胃内的细菌菌落(pcDNA3.1-hspA为28,400±21,600/mm²,pcDNA3.1-hspB为6,800±3,470/mm²)。胃黏膜的组织学分析表明,接种疫苗小鼠的胃炎程度明显低于对照小鼠。这些结果表明,编码幽门螺杆菌热休克蛋白的DNA疫苗可诱导显著的抗幽门螺杆菌免疫反应,以减轻胃黏膜炎症,表明DNA疫苗可能是一种针对人类幽门螺杆菌的新方法。