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

既往研究表明，慢性炎症牙周病变中CD4^+CD45^+T淋巴细胞占主导地位，牙周炎患者血清抗牙龈卟啉单胞菌（P.g） lgG2滴度与临床指标有相关性。为了进一步研究边缘性牙周炎的局部和全身免疫反应，我们检测了外周血T淋巴细胞对P.g.衍生抗原、P.g.声波提取物（p.g.s.e.）和炎症牙周组织中细胞因子产生的增殖反应。25例患者的平均刺激指数（si）为1.63，高于健康志愿者的0.99。7例sii超过2的患者出现明显的增生性反应。在5名表现出高s.i.的健康志愿者中，添加抗cd4和抗II类MHC单克隆抗体可以抑制p.g.s.e.的增殖反应。这表明CD4^+T细胞以更严格的方式识别II类MHC-res中的p.g.g来源抗原，从而导致其增殖。流式细胞术分析显示，在P.g.S.E的刺激下，增殖的T细胞中CD45RA^+CD45RO^+的比例增加。为了更详细地检测CD45异构体的表达，引入了RT-PCR，通过引物在交替剪接的外显子两侧进行检测。扩增的CD45mRNA谱显示，p.g.s.e.刺激的T细胞增加了CD45mRNA的单外显子形式（含第5外显子）和CD45RO异构体mRNA的表达，表明CD45异构体的表达比流式细胞术检测的更为复杂。对牙周炎患者进行牙龈活检，提取RNA，采用RT-PCR检测炎症牙周组织中细胞因子mRNA的表达。T细胞来源的细胞因子、IL-2和IL-4 mrna在牙龈活组织检查的RNA中未检测到，尽管它们在cd3激活的外周血T细胞的RNA中成功扩增。这些结果可能表明，浸润在慢性炎症牙周组织中的T细胞降低了分泌这些细胞因子的能力，并丧失了局部免疫反应的调节功能。进一步的研究是必要的，特别是澄清炎症牙龈中p.g.t细胞激活的细胞因子的产生。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