Immunoresponsive Analysis of Protection against Periodontopathogen

特定的革兰氏阴性菌包括放线菌、牙龈卟啉单胞菌和弯曲杆菌都是牙周病致病菌。本研究的目的是评价放线菌胞丝附着因子的重要性，以及抗胞丝抗体作为附着抑制剂的意义。从放线菌310-a株中制备毛膜抗原。根据菌毛蛋白的氨基酸序列合成寡肽。肽抗原与支链赖氨酸聚合物树脂珠偶联。将肽抗原悬浮在不完全弗氏佐剂乳化的PBS中，用于兔免疫。兔抗血清与A.放线菌comitans 310-a菌毛蛋白的约54kDa蛋白和其他菌毛蛋白反应。该抗血清强烈抑制了a .放线菌菌株对唾液包被混合物的附着。更多的羟基磷灰石珠、颊上皮细胞和成纤维细胞系Gin-1。采用二乙胺解离酶联免疫吸附法测定成人牙周炎患者血清免疫球蛋白G抗体对放线菌和牙龈假单胞菌菌膜抗原的相对亲和力。未感染患者的高滴度血清对菌膜抗原的活性明显高于牙周袋感染患者的高滴度血清。诱导的抗菌膜抗原抗体对放线菌和牙龈假单胞菌感染具有保护作用。制备了兔抗猪链球菌ATCC33238全细胞多克隆血清和小鼠抗猪链球菌表面抗原单克隆抗体。单克隆抗体ATCC33238可与9株直肠球菌中的4株发生反应。该单克隆抗体在体外鉴定出150KDa s层对直梭菌粘附人牙龈成纤维细胞G1细胞的剂量依赖性抑制作用。兔抗血清显著增强了豚鼠腹膜白细胞对直梭菌ATCC33238的吞噬作用，抗s层单克隆抗体有中等增强作用，而抗脂多糖单克隆抗体没有增强作用。在没有抗体的情况下，白细胞暴露于直梭菌细胞的电子显微镜显示出损伤，表明微生物具有白细胞毒性活性。少

Specific gram-negative bacteria including Actinobacillus actinomycetemcomitans, Prophyromonas gingivalis and Campylobacter restus are periodontopathogen. The purpose of this study is to critify the importance of the fimbriae attachment factor of A.actinomycetemcomitants and the significance of anti-fimbrial antibody functions as an attachment inhibitor. Fimbrial antigen was prepared from the A.actinomycetemcomitans 310-a strain. Oligopeptides were synthesized according to the amino acid sequence of the fimbrial protein. The peptide antigen was conjugated with branched lysine polymer resin beads. The peptide antigen was suspended in PBS emulsified with incomplete Freund's adjuvant and used to immunize rabbits. A rabbit antiserum reacted with an approximately 54kDa protein of the fimbriae protein from A.actinomycetemcomitans 310-a and with those of other fimbriated strains. This antiserum atrongly inhibited the attachment of fimbriated A.actinomycetemcomitans strains to saliva-coated hyd … More roxyapatite beads, buccal epithelial cells, and a fibroblast cell line, Gin-1.The relative avidity of serum immunoglobulin G antibody for the fimbria antigens from A.actinomycetemcomitans and P.gingivalis was assessed by diethylamine dissociation enzyme-linked immunosorbent assay in patients with adult periodontitis. High-titer sera from patients not harboring had significantly higher avidities for the fimbria antigen than did high-titer sera from patients with the infections in their periodontal pockets. The elicited antibodies against the fimbria antigen may afford protection against A.actinomycetemcomitans and P.gingivalis infections.Rabbit polyclonal antiserum to whole cells of C.restus ATCC33238 and monoclonal mouse antibodies against surface antigens of the microorganisms were prepared. Monoclonal antibody against 150KDa S-layr protein from C.rectus ATCC33238 reacted with 4 out of 9 C.rectus strains. The monoclonal antibody recognized 150KDa S-layr inhibited dose dependently the C.rectus attachment to human gingival fibroblast, G1 cells, in vitro. Phagocytic killing of C.rectus ATCC33238 by peritoneal leukocytes prepared from guinea pig was significantly enhanced by the rabbit antiserum and moderately enhanced by monoclonal antibody against S-layr, but was not enhanced monoclonal antibody to lipopolysaccharide.Electron micrographs of leukocytes exposed to C.rectus cells on the absence of antibody showed injury, indicating that the microorganisms possesses a leukocyte toxic activity. Less