Immunobiological Studies on the Pathogenic Mechanisms of Periodontal Disease

牙龈卟啉单胞菌（拟杆菌）是一种厌氧的黑色杆状菌，被认为是牙周病的主要病原体。微生物具有毛周菌毛;通过轻轻移液从牙龈卟啉单胞菌菌株381的新鲜全细胞中分离菌毛，并使用DEAESepharose Fast Flow色谱柱进行色谱纯化。牙龈卟啉单胞菌菌毛与佐剂GM-53在脂质体中的口服给药，明显增强了BALB/C小鼠血清IgG反应（IGGI>> IgG_（2b）> IgG_（2a）> IgG_（3））和唾液伊加反应。另一方面，皮下（S。c.）用GM-53注射菌毛也提高了菌毛特异性IgG，随后是血清中的伊加和IgM应答，以及唾液中的伊加和IgG应答。口服免疫的效果不如s。C.在小鼠血清抗体的产生方面。然而，唾液a水平 ...更多信息 注射S. C.与口服免疫小鼠的结果相似。经口免疫BALB/c小鼠，血清中抗菌毛IgG抗体水平维持在较高水平，持续时间约为100 min。初次免疫后7个月。口服给药也诱导并保持在唾液中的菌毛特异性伊加反应至少6个月后的初次immunizations.We已经表明，成人牙周炎（AP）患者表现出血清IgG抗体水平升高牙龈卟啉单胞菌菌毛。亚类反应为IgG 3>> IgG 1> IgG 2>> IgG 4。还观察到一些伊加抗菌毛抗体。IgA 1占主导地位，但显着水平的IgA 2也看到了。然后，我们评估了抗菌毛抗体分泌细胞的数量从外周血单核细胞（PBMC）和从AP受试者的牙龈发炎，通过使用一种新开发的ELISPOT方法。AP患者牙龈单个核细胞（GMC）中含有大量的斑点形成细胞（SFC），其中包括菌毛特异性抗体分泌细胞，其分布规律为IgG>伊加>>IgM，而健康牙龈GMC中SFC数量较少。PBMC中未见菌毛特异性SFC。结果表明，GMC在体外培养时分泌IL-5、IL-6和TGF-β，但不分泌IL-2和IL-4，从而从微生物学和免疫学角度探讨了牙龈卟啉单胞菌在牙周病发病中的作用机制。少

Porphyromonas (Bacteroides) gingivalis, an anaerobic, black-pigmented rod, has been recognized as a major etiologic agent of periodontal disease. The organisms have peritrichous fimbriae ; the fimbriae were detached from fresh whole cells of P. gingivalis strain 381 by gentle pipetting and purified chromatographically using a DEAESepharose Fast Flow column. The purified fimbrial protein gave a single band of 41 KDa.Oral administration of P. gingivalis fimbriae with adjuvant GM-53 in liposomes, but not in Tris-HCI buffer, definitely enhanced the fimbria-specific serum IgG responses (IGGI>>IgG2b>IgG2a>IgG3) and IgA response in saliva of BALB/C mice. On the other hand, subcutaneous (s. c.) injection of fimbriae with GM-53 also raised the fimbria-specific IgG followed by IgA and IgM responses in serum, and both IgA and IgG responses in saliva. Oral immunization was less effective than s. c. injection in terms of the production of serum antibody in the mice. However, the level of salivary a … More ntibody of mice injected s. c. was similar to that of mice immunized orally. High anti-fimbriae IgG in serum were maintained in BALB/c mice immunized orally with fimbriae and GM-53 in liposomes for ca. 7 months after the primary immunizations. Oral administration also induced and held the fimbriaspecific IgA response in saliva for at least 6 months after the primary immunizations.We have shown that patients with adult periodontitis (AP) exhibited elevated serum IgG antibody levels to P. gingivalis fimbriae. The subclass response was IgG3>>IgGI>IgG2>>IgG4. Some IgA anti-fimbriae antibodies were also observed. IgAl predominated, but significant levels of IgA2 were also seen.We then assessed numbers of anti-fimbriae antibody-secreting cells from peripheral blood mononuclear cells (PBMC) and from inflamed gingiva of AP subjects by using a newly developed ELISPOT method. Gingival mononuclear cells (GMC) of AP subjects exhibited high numbers of spot-forming cells (SFC) including fimbriaespecific antibody secreting cells in a pattern of IgG>IgA>>IgM, while low numbers of SFC were seen in GMC from healthy gingiva. No fimbriae-specific SFC were seen in PBMC. It was also found GMC released increased levels of IL-5, IL-6 and TGF-beta but not IL-2 and IL4 when they were grown in vitro.Thus, we have examined pathogenic mechanisms of P. gingivalis in the development of periodontal disease from the veiw points of microbiology and immunology. Less