Development of Immunological Procedure for Prevention of Periodontal Disease Using

预防牙周病的免疫学方法的开发

• 批准号: 08457572
• 负责人: KOGA Toshihiko
• 金额: $ 5.7万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08457572/
• 关键词: Periodontal Disease; Periodontitis; Periodontopathic Bacteria; Monoclonal Antibody; Carbohydrate Antigen; Lipopolysaccharide; 血清型; 熱ショックタンパク質; 抗原; 莢膜多糖

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.

本研究采用单克隆抗体和多克隆抗体分别对牙周病伴放线放线杆菌（Actinobacillusactinomycetemcomitans，Aa）和牙龈卟啉单胞菌（Porphvromonasgingivalis）的细胞表面抗原进行了研究。使用这些抗体，我们获得了下述结果。1）克隆并表征了与Aa Y 4（血清型B）的表型特异性多糖抗原（SPA）的生物合成相关的大基因簇。含有携带该簇的质粒的大肠杆菌产生多糖，该多糖与针对Aa Y 4的SPA的单克隆抗体反应。2）克隆了参与Aa血清型抗原和血清型C抗原合成的基因簇。3)用高压灭菌法从Aa Y 4全细胞中提取SPA，并经DEAE-Sephadex A-25和Sephacryl S-300层析纯化，SPA可诱导人单核细胞释放单核细胞趋化因子和炎性细胞因子。抗SPA单克隆抗体可抑制该因子的释放，而Y 4菌株脂多糖单克隆抗体则不抑制该因子的释放。4）AaY 4及其SPA缺陷型突变体在补体存在下引起人多形核白细胞化学发光反应的能力。白细胞对Y 4菌株的化学发光反应较弱，但对SPA缺陷型菌株的化学发光反应较高。抗SPA单克隆抗体能显著增强白细胞对Y 4菌株的反应。

项目成果

Yoshida,Y.他3名: "Identification of a genetic locus essential for the serotype b-specific antigen synthesis in Actinobacillus actinomycetemcomitans." Infect.Immun.66. 107-114 (1998)

Yoshida, Y. 和其他 3 人：“Actinobacillus actinomycetemcomitans 中血清型 b 特异性抗原合成所必需的遗传位点的鉴定。Infect.Immun.66 (1998)。

Yosheda,Y.他4名: "A novel NDP-6-deoxyhexosyl-4-ulose in the pathway for the synthesis of dTDP-D-fucose." J.Biol.Chem.(印刷中).

Yosheda, Y. 和其他 4 人：“dTDP-D-岩藻糖合成途径中的新型 NDP-6-脱氧己糖基-4-乌糖”（J.Biol.Chem）。

Yoshida, Y.et al.: "Isolation and characterization of chromosomal region containing the gnd from Actinobacillus actinomycetemcomitans." Biochem, Biophys.Res.Commun.230. 220-225 (1997)

Yoshida, Y.et al.：“含有放线放线杆菌伴放线杆菌 gnd 的染色体区域的分离和表征。”

Yoshida, A.et al.: "Isolation and characterization of the danKJ operon from Actinobacillus actinomycetemconitans." DNA Sequence. 8. 93-98 (1997)

Yoshida, A.et al.：“来自 Actinobacillus actinomycetemconitans 的 danKJ 操纵子的分离和表征。”

Yoshida,Y.: "Isolation and characterization of chromosomal region containing the gnd from Actinobacillus actinomycetemcomitans" Biochem.Biophys.Res.Commun.230. 220-225 (1997)

Yoshida,Y.：“含有放线杆菌放线菌 gnd 的染色体区域的分离和表征”Biochem.Biophys.Res.Commun.230。