Clarification of the mechanisms for connective tissue destruction and alveolar bone resorption in periodontal disease by comprehensive analysis of infiltrating T cells using immunological and molecular biological techniques

利用免疫学和分子生物学技术对浸润性 T 细胞进行综合分析，阐明牙周病中结缔组织破坏和牙槽骨吸收的机制

基本信息

批准号：
16390613
负责人：
YAMAZAKI Kazuhisa
金额：
$ 9.09万
依托单位：
Niigata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390613/
关键词：
Regulatory T cells FOXP3 T-cell clone Periodontitis 炎症性サイトカイン抗炎症性サイトカイン mRNA

项目摘要

Periodontitis lesion is characterized by connective tissue destruction and alveolar bone resorption. The lesion contains large numbers of B lymphocytes and plasma cells together with significant numbers of T lymphocytes. We have previously found that the some of the T-cells in the lesion recognize auto-antigens such as heat-shock protein 60 and the response to such antigens play important roles in the pathogenesis of periodontitis. T-cell population can be classified into several distinct subsets functionally as well as phenotypically. In order to elucidate the role of various T-cell subsets in the pathogenesis of periodontal diseases, we comprehensively analyzed the periodontitis lesion-infiltrating T cells using immunological and molecular biological techniques.The results demonstrated that a fraction of CD4^+ T cells expressed both CTLA-4 and CD25. CD4^+CD25^+CTLA-4^+ T cells are thought to be characteristic of natural regulatory T cells (Tr) and the percentage of CD4^+CD25^+ Tr cel … More ls increased with increasing proportions of B-cells relative to T-cells in periodontitis. Gene expression analysis showed that FOXP3, a characteristic marker for CD4^+CD25^+ Tr cells, TGF-β1 and IL-10 were expressed higher in periodontitis than gingivitis. Furthermore, it is demonstrated that another regulatory T cells called NKT cells which has been implicated in periodontitis are activated by CD Id-restricted manner and this NKT cell activation may mediate suppression of auto-reactive T cells.In order to further analyze the role of T cells in the lesion, we established T-cell clones from the gingival tissues of periodontitis patients and examined the effecter function and their gene expression. Most but not all the T-cell clones from gingival tissues expressed mRNA for IFN-γ, IL-4, CD25 and CTLA-4. The frequency of T-cell clones expressing FOXP3 was very high. However, expressions of genes for IL-17 and RANKL, both of which are involved in the bone resorption, were variable among the T-cell clones.Clonal analysis of the T cells clearly demonstrated the presence of T-cell populations reactive to human HSP60 and P. gingivalis GroEL in the peripheral circulation of atherosclerosis patients. These T-cells had been detected in the gingival tissues of periodontitis patients. Furthermore, these HSP60-reactive T cells seemed to be present in atherosclerotic lesions in some patients.In conclusion, periodontitis lesion-infiltrating T cells may be involved in the pathogenesis of not only periodontal diseases but also atherosclerosis. Less

牙周炎病变的特征是结缔组织破坏和牙槽骨吸收，该病变含有大量的 B 淋巴细胞和浆细胞以及大量的 T 淋巴细胞。热休克蛋白 60 等抗原以及对此类抗原的反应在牙周炎的发病机制中发挥着重要作用，T 细胞群可按功能和表型分为几个不同的亚群。为了阐明不同T细胞亚群在牙周病发病机制中的作用，我们利用免疫学和分子生物学技术对牙周炎病灶浸润T细胞进行了全面分析。结果表明，一小部分CD4^+ T细胞同时表达CTLA- 4 和 CD25 CD4^+CD25^+CTLA-4^+ T 细胞被认为是天然调节性 T 细胞 (Tr) 的特征，并且 CD4^+CD25^+ Tr 的百分比。牙周炎中 cel … More ls 随着 B 细胞相对于 T 细胞比例的增加而增加基因表达分析显示，CD4^+CD25^+ Tr 细胞的特征标记物 FOXP3、TGF-β1 和 IL-10 表达较高。此外，已证明与牙周炎有关的另一种称为NKT细胞的调节性T细胞通过CD1d限制性方式被激活。 NKT细胞的激活可能介导自身反应性T细胞的抑制。为了进一步分析T细胞在病变中的作用，我们从牙周炎患者的牙龈组织中建立了T细胞克隆，并检测了效应器功能及其基因表达。来自牙龈组织的大多数（但不是全部）T 细胞克隆表达 IFN-γ、IL-4、CD25 和 CTLA-4 的 mRNA。然而，表达 FOXP3 的 T 细胞克隆的频率非常高。 IL-17 和 RANKL 基因（两者均参与骨吸收）在 T 细胞克隆中存在差异。T 细胞的克隆分析清楚地表明存在对人类 HSP60 和 P 具有反应性的 T 细胞群。在动脉粥样硬化患者的外周循环中检测到这些 T 细胞，此外，这些 HSP60 反应性 T 细胞似乎也存在于牙周炎患者的牙龈组织中。部分患者出现动脉粥样硬化病变。综上所述，牙周炎病灶浸润T细胞不仅可能参与牙周病的发病机制，而且还参与动脉粥样硬化的发病机制。