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

为了表征牙周炎患者浸润牙龈组织的T细胞，我们分别采用RT-PCR和RT-PCR- sscp分析了T细胞因子mrna的表达和克隆性。此外，还研究了牙周病细菌牙龈假单胞菌对外周血T细胞克隆变化的刺激作用。结果表明，PBMC中IFN-y mRNA的平均表达量明显高于牙龈组织。相比之下，牙龈组织中IL-10mRNA的平均表达量高于PBMC。事实上，IL-10mRNA的高表达只出现在7个牙龈组织样本中，大多数样本的表达水平与PBMC相似。IL-12p35 mRNA的平均表达量在牙龈组织和PBMC之间无差异。然而，在16个样本中，有6个样本的牙龈组织中IL-12p40 mRNA的表达高于PBMC，平均表达量差异有统计学意义。IL-13在牙龈组织和PBMC中的平均表达量差异无统计学意义。SSCP分析显示PBMC的涂片模式中条带较少，但在组织样本中大多数Vbeta家族中条带较多。SSCP凝胶上的条带表示在TCR的CDR3区域具有同源核苷酸序列的T细胞克隆。模拟实验表明，刺激后出现了几个新的不同条带，表明克隆积累明显，但大多数情况下不同条带的数量有所减少。这些数据表明，牙龈假单胞菌OM刺激后会发生克隆性改变。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张或积聚。

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.