Kawai T, Ito H O, Sakato N, Okada H
Department of Periodontology, Osaka University Faculty of Dentistry, Suita, Japan.
Clin Diagn Lab Immunol. 1998 Jan;5(1):11-7. doi: 10.1128/CDLI.5.1.11-17.1998.
In the course of long-term infection with Porphyromonas gingivalis in adult periodontitis, a specific antibody response to this organism is generated. We describe a potential novel approach for identifying an immunodominant antigen in human periodontitis patients. First, various monoclonal antibodies (MAbs) were established from mice immunized with crude antigen preparations of P. gingivalis FDC 381. The antigen specificities of these MAbs were compared with those of serum antibodies of 10 periodontitis patients in a competitive enzyme-linked immunosorbent assay. The binding of one MAb (termed PF18) was readily inhibited by sera from all patients but not by sera from healthy volunteers. The antigen recognized by PF18 existed on the cell surface, presumably in the capsule layer, shown by immunoelectron microscopic analysis. Purification of the antigenic substance, termed PF18-Ag, was performed by immunoaffinity chromatography with the MAb. Characterization of PF18-Ag suggested that the epitope was composed of carbohydrates but not peptides and that the substance was different from lipopolysaccharide. Measurement of levels of serum antibody to PF18-Ag better discriminated periodontitis patients from healthy individuals than measurement of antibodies to crude antigen preparations of P. gingivalis. Immunoglobulin G2 was the predominant isotype among the antibodies to PF18-Ag in the patients' sera. These results suggest that PF18-Ag, which is possibly a novel substance, is an important antigenic substance and is potentially useful for the clinical diagnosis of adult periodontitis. The approach that was used would also be relevant to detecting immunodominant antigens of other infectious microorganisms.
在成人牙周炎患者长期感染牙龈卟啉单胞菌的过程中,会产生针对该菌的特异性抗体反应。我们描述了一种在人类牙周炎患者中鉴定免疫显性抗原的潜在新方法。首先,用牙龈卟啉单胞菌FDC 381的粗抗原制剂免疫小鼠,制备了各种单克隆抗体(MAb)。在竞争性酶联免疫吸附试验中,将这些单克隆抗体的抗原特异性与10名牙周炎患者血清抗体的抗原特异性进行了比较。一种单克隆抗体(称为PF18)的结合很容易被所有患者的血清抑制,但不被健康志愿者的血清抑制。免疫电子显微镜分析显示,PF18识别的抗原存在于细胞表面,可能在荚膜层。用该单克隆抗体通过免疫亲和层析法纯化了称为PF18-Ag的抗原物质。PF18-Ag的特性表明,其表位由碳水化合物而非肽组成,且该物质与脂多糖不同。检测血清中针对PF18-Ag的抗体水平比检测针对牙龈卟啉单胞菌粗抗原制剂的抗体能更好地区分牙周炎患者和健康个体。免疫球蛋白G2是患者血清中针对PF18-Ag的抗体中的主要亚型。这些结果表明,PF18-Ag可能是一种新物质,是一种重要的抗原物质,对成人牙周炎的临床诊断可能有用。所采用的方法也可能与检测其他感染性微生物的免疫显性抗原有关。