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Characteristic T cell receptor repertoire and cytokine profile of infiltrating T cells in chronic inflammatory periodontal disease

慢性炎症性牙周病浸润性 T 细胞的特征性 T 细胞受体库和细胞因子谱

基本信息

批准号：
07457454
负责人：
YAMAZAKI Kazuhisa
金额：
$ 4.99万
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：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457454/
关键词：
Periodontitis TCR Cytokine RT-PCR SSCP Porphyromonas gingivalis T cell clonotype

项目摘要

In order to characterize the T cells infiltrating into gingival tissue of periodontitis patients, we analyzed the expression of cytokine mRNAs and clonality of those T cells by using RT-PCR and RT-PCR-SSCP,respectively. Further, the stimulatory effects of P.gingivalis, a periodontopathic bacteria, on the change of peripheral blood T cell clonality was also investigated. The results demonstrated that the mean expression of IFN-y mRNA was significantly higher in PBMC than in gingival tissues. In contrast, the mean expression of IL-10mRNA was higher in gingival tissues than in PBMC.This high expression of IL-10mRNA was in fact, seen in only 7 gingival tissue samples with the majority of samples showing levels similar to PBMC.There was no difference in the mean expression of IL-12p35 mRNA between gingival tissues and PBMC.However, IL-12p40 mRNA was expressed higher in gingival tissues than in PBMC in 6 out of 16 samples with significant difference of mean expression. There was no dofference in the mean expression of IL-13 in gingival tissues and PBMC.SSCP analysis showed smear patterns with few bands in PBMC,but a number of bands in most Vbeta families in tissue samples. A band on the SSCP gel indicate a T cell clone with homogeneous nucleotide sequence in the CDR3 region of the TCR.Simulation experiment demonstrated that several new distinct bands appeared after stimulation, indicating distinct clonal accumulations, although the number of distinct bands decreased in most cases. These data suggest that clonolypic change occurred following stimulation with P.gingivalis OM.These results suggest that T cell clones with type 2 cytokine profile are expanding or accumulating at the site of disease.

为了解牙周炎患者牙龈组织中浸润的T细胞的特征，采用RT-PCR和RT-PCR-SSCP方法分别检测了牙周炎患者牙龈组织中浸润的T细胞的细胞因子mRNA表达和克隆性。此外，还研究了牙周病细菌牙龈卟啉单胞菌对外周血T细胞克隆性变化的刺激作用。结果表明，IFN-γ mRNA在牙周炎患者外周血单个核细胞中的平均表达量明显高于牙龈组织。相比之下，牙龈组织中IL-10 mRNA的平均表达高于PBMC。事实上，IL-10 mRNA的高表达仅出现在7份牙龈组织样本中，大多数样本的水平与PBMC相似。牙龈组织和PBMC之间IL-12 p35 mRNA的平均表达没有差异。然而，16例中有6例牙龈组织IL-12 p40 mRNA表达高于PBMC，两者平均表达量差异有统计学意义。SSCP分析结果显示，牙龈组织中IL-13的表达量与PBMC无显著性差异，而组织中大多数Vbeta家族的表达量明显高于PBMC。SSCP凝胶上的一条条带表明TCR的CDR 3区有同源的核苷酸序列，模拟实验表明刺激后出现了几条新的明显条带，表明有明显的克隆积累，尽管在大多数情况下明显条带的数量减少。这些数据表明，在用牙龈卟啉单胞菌OM刺激后发生了克隆形成变化。这些结果表明，具有2型细胞因子谱的T细胞克隆在疾病部位扩增或积累。