Analysis of antigen receptor genes of T cells specific for the periodontal disease

我们以前的研究使用RT-PCR-SSCP分析表明，高克隆性T细胞浸润牙周炎病变类似于自身免疫性疾病，如类风湿性关节炎的炎性病变。考虑到牙周袋中复杂的微生物植物群，推测T细胞识别与宿主成分同源的细菌抗原。由于热休克蛋白60是显着的免疫原性，尽管其高度的进化保守性，T细胞和抗体的反应，热休克蛋白60已被报道在各种炎症条件。为了阐明hsp 60 s在牙周炎T细胞反应中的作用，我们检测了牙周炎患者和对照组外周血单个核细胞（PBMC）在重组人hsp 60和牙龈卟啉单胞菌（P.gingivalis）GroEL刺激下的增殖反应以及T细胞的细胞因子谱和克隆性。此外，nu 关于我们 比较hsp 60反应性PBMC和牙周炎病变浸润性T细胞之间T细胞受体β链CDR 3区域内的cleotide序列。牙周炎患者PBMC对人hsp 60的增殖反应显著高于对照组，但对牙龈卟啉单胞菌GroEL的增殖反应不显著。CDR 3区的核苷酸序列分析清楚地表明，hsp 60反应性T细胞克隆和牙周炎病变浸润性T细胞表达相同的受体，表明hsp 60反应性T细胞在牙周炎病变中积累。细胞因子谱分析表明，hsp 60反应性PBMC在牙周炎患者中产生显著水平的IFN-γ，而牙龈卟啉单胞菌GroEL不诱导任何向1型或2型的偏移。结果提示，牙周炎患者牙周致病菌感染后，自身hsp 60反应性T细胞活化，产生1型细胞因子，刺激巨噬细胞产生促炎性细胞因子，导致牙周组织破坏。少

Our previous studies using RT-PCR-SSCP analysis demonstrated the high clonality of T cells infiltrating into periodontitis lesion similar to that in inflammatory lesion of autoimmune diseases such as rheumatoid arthritis. Considering that the complex microbial flora in the periodontal pocket, it was speculated that T cells are recognizing antigens of bacteria which are homologous to host components. Because heat shock protein 6Os are remarkably immunogenic, despite their high degree of evolutionary conservation, both T cell and antibody responses to hsp60 have been reported in a variety of inflammatory conditions. In order to clarify the role of hsp60s on the T cell response in periodontitis, we examined the proliferative response of peripheral blood mononuclear cells (PBMC) as well as cytokine profile and clonality of T cells upon stimulation with recombinant human hsp6O and Porphyromonas gingivalis (P.gingivalis) GroEL from periodontitis patients and control subjects. Furthermore, nu … More cleotide sequences within the CDR3 region of the T cell receptor β-chain was compared between hsp60-reactive PBMC and periodontitis lesion-infiltrating T cells. In periodontitis patients significantly higher proliferative response of PBMC to human hsp60 but not to P.gingivalis GroEL than in control subjects. Analysis of the nucleotide sequences within the CDR3 region clearly demonstrated that hsp60-reactive T cell clones and periodontitis lesion-infiltrating T cells expressed the same receptors suggesting that hsp60-reactive T cells accumulated in periodontitis lesion. The analysis of cytokine profile demonstrated that hsp60-reactive PBMC produced significant level of IFN-γ in periodontitis patients, whereas P.gingivalis GroEL did not induce any skewing towards type1 or type2. In control subjects no significant expression of IFN-γ or IL-4 was induced upon stimulation with either hsp60 or P.gingivalis GroEL.These results suggest that in periodontitis patients infection with periodontopathic bacteria may induce the activation of self hsp60-reactive T cells with type1 cytokine profile resulting the stimulation of macrophage to produce proinflammatory cytokines leading to the tissue destruction. Less