Molecular and immunological analysis of periodontopathic bacterial attachment factors

人类牙周病是由布拉姆阴性菌引起的慢性传染病。许多革兰氏阴性菌已从人类口腔中分离出来。放线菌是嗜碳棒，与青少年牙周炎有关。我们成功地生产了微生物菌毛合成肽疫苗。我们发现用编码白介素4和霍乱毒素的质粒DNA的肽疫苗免疫可诱导唾液IgA抗体的吸附。该疫苗可诱导对定植的保护作用。牙龈卟啉单胞菌是一种厌氧黑色色素棒，已被认为是成人牙周炎的致病生物。许多毒力因素已牵连到致病性牙龈假单胞菌。由微生物产生的精氨酸特异性半胱氨酸蛋白酶（gpA和gpB）被怀疑是毒力因子，并参与中断宿主防御机制以及破坏牙周结缔组织。为了诱导对牙龈假单胞菌的保护性免疫应答，我们成功构建了argA DNA疫苗。用携带gpA质粒DNA的基因枪皮下免疫BALBc小鼠。采用酶联免疫吸附法测定抗牙龈假单胞菌的抗体反应。rgpA DNA疫苗诱导了高水平的血清抗牙龈假单胞菌抗体。rgpA DNA疫苗免疫小鼠的血清降低了gpA和gpB的蛋白水解活性，抑制了牙龈卟啉卟啉菌与胶原海绵的结合。此外，血清有效地降低了牙龈卟噬菌细胞的血凝活性，表明生物体的血凝活性与gpA有关。我们在小鼠模型中发现，用rgpA DNA疫苗免疫的小鼠对密集的牙龈假单胞菌W50攻击具有抗性。这些结果表明，rgpA DNA疫苗诱导了针对该酶的特异性抗体，该疫苗可以赋予对牙龈假单胞菌感染的保护性免疫。少

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