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Induction of mucosal immune responses by DNA vaccine encoding Porphyromonas gingivalis fimbriae

编码牙龈卟啉单胞菌菌毛的 DNA 疫苗诱导粘膜免疫反应

基本信息

批准号：
12671769
负责人：
KAWABATA Shigetada
金额：
$ 2.37万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

项目摘要

For the development of vaccines against oral and pharyngeal pathogens invading the mucosal epithelia, both secretory and serum IgA/IgG antibodies and cytotoxic T lymphocytes have been induced. We employed a novel approach, targeted salivary gland (TSG) immunization, using a plasmid pcDNA3/fimA coding for Porphyromonas gingivalis fimbriae. Expression of subunit protein, fimbrillin, was observed in eukaryotic cells growing in vitro following transfection, with pcDNA3/fimA. In this study, we obtained good humoral and cell-mediated immune responses in BALB/c mice by TSG administration using the above mentioned DNA vaccine. The production 6f fimbriae-specific IgA and IgG antibodies in saliva and serum IgG antibody was significantly stimulated by TSG immunization. Injection of DNA vaccine into salivary gland elicited high level production of antigen-specific IgG antibody, similar to that induced following intramuscular immunization. The major IgG subclass that recognized fimbriae was IgG2a in serum from pcDNA3/fimA immunized mice. When mononuclear cells from salivary glands^were analyzed by RT-PCR, higher levels of Th2 cytokine-specific mRNA were seen in the immunized group than non-immunized group. In addition, TSG DNA immunization resulted in the generation of antigen-specific CTL in spleen. These results indicate that TSG immunization with plasmid DNA may represent a genetic immunization strategy against infection by oral and pharyngeal pathogens that may invade local, mucosal surfaces.

为了开发针对口腔和咽部病原体侵入粘膜上皮的疫苗，已经诱导了分泌型和血清中的IgA/Ig G抗体和细胞毒性T淋巴细胞。我们采用了一种新的方法，即靶向唾液腺(TSG)免疫，使用编码牙龈卟啉单胞菌菌毛的质粒pcDNA3/FIMA。将pcDNA3/FIMA导入体外培养的真核细胞中，观察到亚基蛋白fimbrin的表达。在本研究中，我们用上述DNA疫苗TSG免疫BALB/c小鼠，获得了良好的体液免疫和细胞免疫应答。TSG免疫可显著刺激小鼠唾液和血清中6f菌毛特异性IgA和Ig G抗体的产生。唾液腺注射DNA疫苗可诱导高水平的抗原特异性免疫球蛋白抗体产生，与肌肉注射免疫后产生的抗体相似。PcDNA3/FIMA免疫小鼠血清中识别菌毛的主要免疫球蛋白亚类为IgG2a。用RT-PCR分析唾液腺单个核细胞，免疫组Th2细胞因子特异性mRNA的表达水平高于未免疫组。此外，TSG DNA免疫可在小鼠脾内产生抗原特异性CTL。这些结果表明，用质粒DNA进行TSG免疫可能代表了一种针对口腔和咽部病原体感染的遗传免疫策略，这些病原体可能侵袭局部的粘膜表面。