Molecular and immunological analysis of periodontopathic bacterial attachment factors

人类牙周病是由BRAM阴性菌引起的慢性感染性疾病。从人类口腔中分离出了许多革兰氏阴性细菌。伴生放线杆菌是一种嗜碳酸杆菌，与青少年牙周炎有关。我们成功地生产出了该微生物菌毛的合成肽疫苗。我们发现，用编码白介素4和霍乱毒素的DNA多肽疫苗免疫小鼠，可诱导产生黏附抑制性唾液IgA抗体。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素杆菌，已被认为是成人牙周炎的致病微生物。许多毒力因子被认为与牙周炎的致病性有关。由该微生物产生的精氨酸特异性半胱氨酸蛋白酶(gpa和gpb)是可疑的毒力因子，参与干扰宿主防御机制和PEN…。更多酸蚀和破坏牙周结缔组织。为了诱导对牙龈假单胞菌的保护性免疫反应，我们成功构建了Arga DNA疫苗。用携带GPA的基因枪对BALBc小鼠进行皮内免疫。用酶联免疫吸附试验检测抗牙周炎杆菌抗体反应。RgpA DNA疫苗可诱导高水平的抗牙周血吸虫抗体。RgpA DNA疫苗免疫的小鼠血清降低了GPA和GPB的蛋白分解活性，并抑制了牙龈假单胞菌与胶原海绵的结合。此外，该血清还能有效地降低牙龈假单胞菌细胞的血凝活性，表明该菌的血凝活性与GPA有关。我们用小鼠模型发现，用rgpA DNA疫苗免疫的小鼠对牙龈假单胞菌W50的强化攻击具有抵抗力。这些结果表明，rgpA DNA疫苗诱导产生了针对该酶的特异性抗体，该疫苗可以产生保护性免疫，以抵抗牙龈假单胞菌感染。较少

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