Mechanisms of the Periodontal Tissue Destruction by Periodontopathogenic Bacteria

一些由牙周病细菌产生的蛋白酶被认为是牙周病的潜在致病因素。我们已经确定牙龈卟啉单胞菌可能是成人牙周炎的一种牙周致病菌，它能产生几种类型的蛋白酶。本研究描述了这些毒力因子对牙周组织破坏的机制。从培养上清和囊泡中分离纯化了3种对BApNA有活性的蛋白酶（Pase-B、Pase-C和Pase-S）。分别制备针对Pase-S和Pase-C的多克隆和单克隆抗体（mab - ps,mAb-PC），并测定这些蛋白酶的免疫学特性。Pase-S和Pase-B与mab - ps反应，但与mAB-PC不反应；Pase-C与mAB-PC反应，但与mab - ps不反应。mAB-PC仅与牙龈假单胞菌发生反应，而与其他可能产生BApNA活性的牙周病致病菌（如Bac）没有反应。这些蛋白酶的生物活性也各不相同，即I型胶原仅对Pase-C敏感，而IV型胶原被所有蛋白酶破坏。根据底物特异性，Pase-C类似于胰蛋白酶，而Pase-B和Pase-S更类似于clostripain。病原菌的聚集活性通常与病原菌对宿主组织的粘附有关。为了估计牙龈假单胞菌可能产生的毒力因子的生物活性之间的关系，以细胞聚集活性为因变量，以BApNA水解酶、精氨酸羧肽酶和精氨酸脱亚胺酶3种精氨酸代谢酶活性为解释变量，采用多元回归分析进行统计分析。这些统计分析结果表明，BApNA水解活性与细胞聚集活性呈正相关，而精氨酸脱亚胺酶活性与细胞聚集活性呈负相关。这些结果表明Pase-C在牙周组织的粘附和破坏中起着重要的作用。少

Some kinds of proteases produced by periodontopathogenic bacteria are considered to be the putative viruent factor of periodontal disease. We have already determined that Porphyromonas gingivalis, a probalbe periodontopathogenic bacterium in adult periodontitis, produces several types of proteases. This study describes the mechanisms of the periodontal tissue destruction by these virulent factors.Three kinds of proteases (Pase-B,Pase-C and Pase-S) which were active on BApNA were isolated and purified from the culture supernatant and vesicles. Polyclonal and monoclonal antibodies were prepared against Pase-S and Pase-C,respectively (pAb-PS,mAb-PC) and the immunological characteristics of these proteases were determined. Pase-S and Pase-B reacted with pAB-PS,but did not with mAB-PC,while Pase-C reacted with mAB-PC but did not with pAB-PS.mAB-PC only reacted with strains of P.gingivalis but did not with other putative periodontopathogenic bacteria which produced BApNA activity such as Bac … More teroides forsUthus and Treponema denticola. Different biological activities were also found in these proteases, that is, type I collagen was sensitive only for Pase-C,but type IV collagen was destroyed by all of these proteases. On the basis of substrate specificity, Pase-C resembles trypsin but Pase-B and Pase-S rather resemble clostripain.As the aggregating activity of the pathogenic bacterium is generally correlated with the bacterial adhesion to host tissues. In order to estimate the relationship between biological activities of possible virulent factors produced by P.gingivalis, statistical analyzes were performed by multiple regression analysis using the cell aggregating activity as the dependent variable and three arginine metabolizing enzyme activities, BApNA hydrolyzing enzyme, arginine carboxypeptidase and arginine deiminase, as explanatory variables. The results of these statistic analyzes suggested that BApNA hydrolyzing activity had the strongest positive relationship to the cell aggregating activity, while arginine deiminase activity showed a negative relationship. These results strongly suggest that Pase-C plays important roles in the adhesion to the periodontal tissues and their destruction. Less