Molecular and immunological analysis of periodontopathic bacterial attachment factors

牙周病细菌附着因子的分子和免疫学分析

基本信息

批准号：
11470381
负责人：
OKUDA Katsuji
金额：
$ 6.78万
依托单位：
TOKYO DENTAL COLLEGE
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

项目摘要

Human periodontal diseases are chronic infectious desease by bram-negative bacteria. A number of gram-negative bacteria have been isolated from the human oral cavity. Actinobacillus actinomycetemcomitans is capnophilic rod that has been implicated in juvenile periodontitis. We succeeded to produce synthetic peptide vaccine of fimbriae of the microorganism. We found that immunization with the peptide vaccine with plasmid DNA which coding interleukin 4 and cholera toxin induced attacment inhibitory salivary IgA antibody. The vaccine may induce the protective action for the colonization. Porphyromonas gingivalis is an anaerobic black-pigmented rod that has been implicated as a causative organism in adult peridontitis. A number of virulence factors have been implicated in pathogenicity of P. gingivails. Arginine specific cysteine proteinases (gpA and gpB) produced by the microorganism are suspected virulence factors and are involved in interrupting host defense mechanisms as well as in pen … More etrating and destroying periodontal connective tissues. To induce a protective immune response against P. gingivalis, we succeeded to construct an argA DNA vaccine. BALBc mice were immunized intradermally by Gene Gun with plasmid DNA carrying gpA. Antibody responses against P. gingivalis were determined by an enzyme-linked immunosorbent assay. The rgpA DNA vaccine induced high levels of serum antibodies aginst P. gingivalis. Sera from the rgpA DNA vaccine-immunized mice diminished the proteolytic activity of gpA and gpB and inhibited the binding of P. gingivalis to a collagen sponge. Moreover, the sera effectively reduced the hemagglutinating activity of P. gingivalis cells, indicating that hemagglutinio activity of the organism is associated with gpA. We found with a murine model that mice immunized with rgpA DNA vaccine were resistant to an intensive P. gingivalis W50 challenge. These results suggest that the rgpA DNA vaccine induced specific antibodies against the enzyme and that this vaccine could confer protective immunity against P. gingivalis infection. Less

人类牙周病是一种由革兰氏阴性菌引起的慢性传染病，从人类口腔中分离出多种革兰氏阴性菌，它是与幼年牙周炎有关的嗜二氧化碳杆菌。我们发现用编码白细胞介素4和霍乱毒素的质粒DNA的肽疫苗进行免疫。疫苗可诱导牙龈卟啉单胞菌定植的保护作用，牙龈卟啉单胞菌是一种厌氧黑色素杆状细菌，已被认为是成人牙周炎的致病微生物。牙龈产生的精氨酸特异性半胱氨酸蛋白酶（gpA 和 gpB）。微生物被怀疑是毒力因子，参与破坏宿主防御机制以及破坏牙周结缔组织。为了诱导针对牙龈卟啉单胞菌的保护性免疫反应，我们成功构建了 argA DNA 小鼠。用携带 gpA 的质粒 DNA 进行皮内免疫，通过酶联免疫吸附测定测定针对牙龈卟啉单胞菌的抗体反应。 rgpA DNA 疫苗免疫小鼠血清中诱导出高水平的抗牙龈卟啉单胞菌抗体，降低了 gpA 和 gpB 的蛋白水解活性，并抑制了牙龈卟啉单胞菌与胶原海绵的结合，此外，血清还有效降低了血凝活性。牙龈卟啉单胞菌细胞，表明该生物体的血凝活性与 gpA 相关，我们在小鼠免疫的小鼠模型中发现。使用 rgpA DNA 疫苗对牙龈卟啉单胞菌 W50 的强烈攻击具有抵抗力。这些结果表明，rgpA DNA 疫苗诱导了针对该酶的特异性抗体，并且该疫苗可以赋予针对牙龈卟啉单胞菌感染的保护性免疫力。