Functional analyzes of peripheral blood and locally infiltrated T cells in periodontitis patients.

以往的研究表明，慢性牙周炎患者外周血中CD 4 ^+ CD 45 ^+T淋巴细胞占优势，血清中抗牙龈卟啉单胞菌（P.g.）与牙周炎患者的临床指标相关。为了进一步研究边缘性牙周炎的局部和全身免疫反应，研究了外周血T淋巴细胞对P.g.-衍生抗原，P.g.超声提取物（P.g.S.E.）并检测炎症牙周组织中细胞因子的产生。刺激指数（S.I.）25例患者的平均值为1.63，高于健康志愿者的0.99。7例患者对P.g.S.E.显示出显著的增殖反应，S.I.2以上。在5名表现出高S.I.的健康志愿者中，通过加入抗CD 4和抗II类MHC单克隆抗体抑制对P.g.S.E.的增殖反应。这表明CD 4 ^+T细胞识别P.g.- II类MHC-res衍生抗原 关于我们 的方式，导致其扩散。流式细胞术分析显示，在P.g.S.E.刺激下，增殖T细胞中CD 45 RA ^+ CD 45 RO ^+的比例增加。为了详细检测CD 45亚型的表达，引入了通过可变剪接外显子侧翼引物的RT-PCR。扩增的CD 45 mRNA的谱显示P.g.S.E.-刺激后的T细胞表达CD 45 RO亚型mRNA，同时也表达含第5外显子的单外显子形式的CD 45 mRNA，表明CD 45亚型的表达比流式细胞术检测的更为复杂。从牙周炎患者牙龈组织中提取RNA，用RT-PCR方法检测炎症牙周组织中细胞因子mRNA的表达，结果显示，T细胞源性细胞因子IL-2和IL-4 mRNA在牙龈组织中未检测到，但在外周血CD 3活化T细胞中却能扩增到。这些结果可能表明，慢性炎症牙周组织中浸润的T细胞降低了分泌这些细胞因子的能力，丧失了局部免疫反应的调节功能。进一步的研究将是必要的，特别是澄清细胞因子的生产P.g.发炎牙龈中的活化T细胞。少

Previous studies demonstrated that CD4^+CD45^+T lymphocytes predominated in chronically inflamed periodontal lesion and the serum lgG2 titer against Porphyromonas gingivalis (P.g.) exhibited corelation with clinical index in periodontitis patients. To further examine the local and general immune responses involved in marginal periodontitis, proliferative responses of peripheral blood T lymphocytes against P.g.-derived antigens, P.g. sonicate extract (P.g.S.E.) and cytokine production in inflamed periodontal tissue was examined. The average of stimulation index (S.I.) in 25 patients was 1.63, which is higher than 0.99 obtained from healthy volunteers. Seven patients showed remarked proliferative responses to P.g.S.E.with over S.I.2. In the 5 healthy volunteers who exhibited high S.I., proliferative response to P.g.S.E.was inhibited by the addition of anti-CD4 and anti-class II MHC monoclonal antibodies. This suggested that CD4^+T cells recognized P.g.-derived antigen in class II MHC-res … More tricted manner, resulting in their proliferation. Flow cytometric analysis showed that the proportion CD45RA^+CD45RO^+ was increased among proliferating T cells by the stimulation of P.g.S.E.. To detect CD45 isoform expression in detail, RT-PCR by the primers flanking the alternatively spliced exons was introduced. The profile of amplified CD45 mRNA revealed that P.g.S.E.-stimulated T cell increased the expression of single exon form of CD45mRNA,containing 5th exon, as well as CD45RO isoform mRNA,indicating more complicated expression of CD45 isoform than that detected by flow cytometry. Gingival biopsies from periodontitis patients, where RNA was extracted to examine the expression of cytokine mRNA in inflamed periodontium by RT-PCR.T cell-derived cytokine, IL-2 and IL-4 mRNAs were not detected in RNA from gingival biopsies althogh they were successfully amplified in RNA from CD3-activated peripheral blood T cells. These results may suggest that infiltrating T cells in chronically inflamed periodontium decrease the ability to secret those cytokines and loss the regulatory functions in local immune responses. Further investigation would be necessary particulary for clarifying cytokine production by P.g.-activated T cells in inflamed gingiva. Less