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Development of effective immunization by DNA vaccine against iron-acquisition protein of periodontopathic bacteria

DNA疫苗开发针对牙周病细菌铁获取蛋白的有效免疫

基本信息

批准号：
14370694
负责人：
SHIZUKUISHI Satoshi
金额：
$ 8.38万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

Lysine-specific cysteine proteinase (KGP) of Porphyromonas gingivalis, which is a prominent periodontopathic bacterium, is considered to play an important role in its pathogenesis, since it possesses protease activity as well as hemoglobin-binding activity. Previously, we reported that immunization of mice by injection of DNA vaccine against KGP into the muscle induced specific anti-KGP IgG antibodies in serum and that the DNA vaccine prevented inflammatory responses and prolonged the survival rate of the immunized mice against P gingivalis infection. The aim of this study was to investigate whether humoral immunity and cell-mediated immunity could be introduced when mice were immunized by injection of DNA vaccine into muscle or by oral administration of DNA vaccine-harboring avirulent bacterium that parasitizes in host cells; consequently to develop effective immunization. We constructed kgp-DNA vaccine (pCMV/kgp) for oral administration and transformed the plasmid into Salmonella typ … More himuriuin SL7207, which is an avirulent bacterium and was used as a vehicle for the DNA vaccine. Mice were immunized orally with the bacterium harboring the plasmid encoding kgp-DNA vaccine weekly for a total of four inoculations. Following the immunization, it was shown that specific anti-KGP IgG antibodies in serum were clearly induced by ELISA. Although induction of anti-KGP IgG antibodies in serum was observed when mice were immunized by injection of the DNA vaccine into muscle, the titers were lower than those when mice were orally immunized. Moreover, anti-KGP IgA antibodies in saliva were also introduced by immunization by oral administration of the DNA vaccine, while those were not observed when mice were immunized by intramuscular injection. We are now working on development of effective immunization methods and ion introduction of cell-mediated immunity and mucosal immunity. The present study indicates that immunization of kgp-DNA vaccine by oral administration may be a useful method to prevent periodontitis. Less

牙龈卟啉单胞菌是一种重要的牙周病细菌，其赖氨酸特异性半胱氨酸蛋白酶（KGP）被认为在其发病机制中发挥着重要作用，因为它具有蛋白酶活性以及血红蛋白结合活性。此前，我们报道了通过将针对KGP的DNA疫苗注射到肌肉中来免疫小鼠，在血清中诱导了特异性抗KGP IgG抗体，并且DNA疫苗预防了炎症反应并延长了免疫小鼠针对牙龈卟啉单胞菌感染的存活率。本研究的目的是探讨通过肌肉注射DNA疫苗或口服寄生在宿主细胞内的携带无毒力细菌的DNA疫苗来免疫小鼠，是否可以引入体液免疫和细胞介导的免疫。从而开展有效的免疫接种。我们构建了用于口服给药的 kgp-DNA 疫苗（pCMV/kgp），并将质粒转化为沙门氏菌型 Himuriuin SL7207，这是一种无毒细菌，用作 DNA 疫苗的载体。每周用携带编码kgp-DNA疫苗的质粒的细菌口服免疫小鼠，总共接种四次。免疫后，ELISA结果显示，血清中明显诱导出特异性抗KGP IgG抗体。虽然当通过将DNA疫苗注射到肌肉中来免疫小鼠时观察到血清中抗KGP IgG抗体的诱导，但是滴度低于小鼠口服免疫时的效价。此外，通过口服DNA疫苗免疫也引入了唾液中的抗KGP IgA抗体，而通过肌肉注射免疫小鼠时未观察到这些抗体。我们现在正在致力于开发有效的免疫方法以及细胞介导免疫和粘膜免疫的离子引入。本研究表明，口服kgp-DNA疫苗免疫可能是预防牙周炎的有效方法。较少的