Immunoresponsive Analysis of Protection against Periodontopathogen

特殊的革兰氏阴性细菌包括伴生放线杆菌、牙龈预防单胞菌和静止弯曲杆菌是牙周病的致病菌。本研究的目的是评价放线菌伴生菌菌毛附着因子的重要性，以及抗菌毛抗体作为附着抑制因子的意义。从伴放线放线菌310-a菌株中制备菌毛抗原。根据菌毛蛋白的氨基酸序列合成寡肽。将多肽抗原与支化赖氨酸聚合物树脂小球偶联。用不完全弗氏佐剂乳化的PBS悬浮肽抗原免疫兔。兔抗血清可与放线菌310-a菌毛蛋白的约54 kDa蛋白及其它菌毛蛋白反应。该抗血清可显著抑制伴生放线菌菌毛与唾液包被的HYD…的黏附。用二乙胺解离酶联免疫吸附试验检测了成人牙周炎患者血清免疫球蛋白G抗体对放线菌菌毛抗原和牙周炎菌毛抗原的相对亲和力。没有感染的患者的高滴度血清对菌毛抗原的亲和力显著高于牙周袋感染患者的高滴度血清。所诱导的抗放线菌菌毛抗原抗体可能对伴生放线菌和牙龈假单胞菌的感染具有保护作用。ATCC33238株150 KDa S-layR蛋白的单抗可与9株直肠弯曲菌中的4株发生反应。识别150 KDa S-LayR的单抗在体外呈剂量依赖性地抑制直肌与人牙龈成纤维细胞G1细胞的黏附。兔抗血清能显著增强豚鼠腹膜白细胞对直肠弯曲菌ATCC33238的吞噬杀伤作用，抗S-LayR单抗对其有中等程度的增强作用，而抗脂多糖单抗不能增强这一作用。较少

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