Development of Immunological Procedure for Prevention of Periodontal Disease Using

本研究对牙周病放线杆菌和牙龈假单胞菌的细胞表面抗原进行了单抗和多克隆抗体的检测。利用这些抗体，我们获得了如下结果：1)克隆并鉴定了与AAY4(b型)血清型特异性多糖抗原(SPA)生物合成相关的大基因簇。(2)克隆了AA型a型和c型A型抗原合成相关基因簇。3)高压灭菌提取的AA Y4全细胞SPA，经DEAE-Sephadex A-25和Sephacryl S-300层析纯化后，可诱导人单核细胞释放趋化因子和炎性细胞因子。该因子的释放可被抗SPA的单抗所抑制，但不能被Y4菌株的脂多糖单抗所抑制。4.4)在补体存在的情况下，AA Y4及其SPA缺陷变异体能够诱发人多形核白细胞的化学发光反应。白细胞对Y4菌株的化学发光反应较弱，但对SPA缺陷变异体的化学发光反应较高。抗SPA的单抗可显著增强白细胞对Y4株的反应。

In this study, monoclonal antibodies and polyclonal antibodies against cell-surface antigens of periodontopathic Actinobacillus actinomycetemcomitans (Aa) and Porphvromonas gingivalis. Using these antibodies, we have obtained the results described below.1) A large gene cluster associated with the biosynthesis of the serotype-specific polysaccharide antigen (SPA) of Aa Y4 (serotype b) was cloned and characterized. Escherichia coli containing a plasmid carrying this cluster, produced a polysaccharide which reacted with a monoclonal antibody directed against the SPA of Aa Y4.2) Gene clusters involved in the synthesis of the serotype a antigen and the serotype c antigen of Aa were cloned. 3) SPA extracted from whole cells of Aa Y4 by autoclaving and purified by chromatography on DEAE-Sephadex A-25 and Sephacryl S-300 induced the release of monocyte chemotactic factor and inflammatory cytokines by human monocytes. The release of the factor was inhibited by monoclonal antibodies against SPA, but not by monoclonal antibodies of lipopolysaccharide from strain Y4.4) The ability of Aa Y4 and its SPA-defective variants to elicit the chemiluminescence response of human polymorphonuclear leukocytes in the presence of complement. The chemiluminescence response of leukocytes to strain Y4 was weak, but that to the SPA-defective variants was high. Monoclonal antibodies against SPA enhanced significantly the response of leukocytes to strain Y4.