Host responses against heat shock proteins from periodontopathogenic bacteria

宿主对牙周病原菌热休克蛋白的反应

基本信息

批准号：
10470447
负责人：
HINODE Daisuke
金额：
$ 6.46万
依托单位：
The University of Tokushima
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

The metabolism of oral microorganisms is largely influenced by the changing environment which also lead to the synthesis of stress proteins such as heat shock proteins (HSPs). HSPs belong to some families of highly conserved homology. We focused on the periodontal disease and HSPs as periodontopathogen because these proteins were immunodominant antigens. Our results indicated that the GroEL-like protein, belonging to HSP60 family, from periodontopathogens possessed epitope cross-reactive with that from human origin. The nucleotide sequence was determined directly using the PCR product corresponding to the Campylobacter rectus GroEL-like protein, and analyzed. By the comparison of the alignment of the deducted amino acid sequence with those of human HSP60, the similarity throughout its length was confirmed. It is possible that high homology of HSPs within the same family and their common epitopes with host molecule lead to autoimmune responses.The native GroEL-like protein from C.rectus and Actinobacillus actinomycetemcomitans possessed biological activities. The C.rectus GroEL-like protein stimulated both interleukin-6 (IL-6) and IL-8 secretion by a confluent monolayer of human gingival fibroblast in their culture supernatant. During the 22h-incubation, the amounts of IL-6 and IL-8 were increased by 5.4- and 3.5-fold, respectively. A.actinomycetemcomitans GroEL-like protein had a significant cytotoxic effect on both periodontal ligament epithelial cells and HaCaT epithelial cells grown in vitro at higher concentrations. On the other hand, we determined the external localization of A.actinomycetemcomitans GroEL-like protein with specific antibodies using immunoelectron microscopy. These results demonstrated that the A.actinomycetemcomitans GroEL-like protein and the C.rectus GroEL-like protein possessed biological activities influencing on the host cells, therefore, these proteins could play important roles in periodontal disease.

口腔微生物的代谢很大程度上受到环境变化的影响，这也会导致应激蛋白的合成，例如热休克蛋白（HSP）。 HSP 属于一些高度保守的同源性家族。我们重点关注牙周病和作为牙周病原的热休克蛋白，因为这些蛋白质是免疫显性抗原。我们的结果表明，来自牙周病原菌的属于HSP60家族的GroEL样蛋白具有与人类来源的表位交叉反应的表位。使用对应于直肠弯曲杆菌GroEL样蛋白的PCR产物直接确定核苷酸序列并进行分析。通过将推导的氨基酸序列与人HSP60的氨基酸序列进行比对，证实了其整个长度的相似性。同一家族内的HSPs的高度同源性及其与宿主分子的共同表位可能导致自身免疫反应。来自直立杆菌和伴放线放线杆菌的天然GroEL样蛋白具有生物活性。 C.ectus GroEL 样蛋白刺激培养物上清液中汇合的单层人牙龈成纤维细胞分泌白细胞介素 6 (IL-6) 和 IL-8。在22小时的孵育期间，IL-6和IL-8的量分别增加了5.4倍和3.5倍。 A.actinomycetemcomitans GroEL样蛋白在较高浓度下对体外生长的牙周膜上皮细胞和HaCaT上皮细胞均具有显着的细胞毒作用。另一方面，我们使用免疫电子显微镜用特异性抗体确定了 A.actinomycetemcomitans GroEL 样蛋白的外部定位。这些结果表明A.actinomycetemcomitans GroEL-like蛋白和C.ectus GroEL-like蛋白具有影响宿主细胞的生物活性，因此，这些蛋白在牙周疾病中可能发挥重要作用。