Detection of periodontal disease activity by T cell receptor repertoire and cytokine profile of infiltrating T cells

为了明确牙周炎患者浸润牙龈组织的T细胞特征，采用免疫组织学方法对T细胞受体库和细胞因子谱进行了研究。在完成初始治疗后进行牙周手术时获得牙周炎组织。作为对照的牙龈炎组织也从非牙周炎病变中获得。用碱性磷酸酶和抗碱性磷酸酶法对每个标本的连续冷冻切片进行TCR V基因产物（Valpha2、Vbeta5.2-3、Vbeta5.3、Vbeta5.1、Vbeta6.7、Vbeta8、Vbeta12）和细胞因子（IL-2、ifn - γ、IL-4、IL-6）的免疫染色。通过将每个阳性细胞数除以CD3阳性细胞总数来计算相对百分比。此外，为了比较牙龈组织和循环池，用流式细胞术分析两组外周血单个核细胞的TCR库。各TCR - V基因产物的表达模式在炎症牙龈中表现出个体差异，但呈现出一致的表达模式。Vbeta5亚家族和Vbeta6.7是牙龈中常用的基因库，而vbeta2和Vbeta8亚家族在大多数情况下低表达。此外，与自体外周血相比，牙龈组织中TCR V基因产物的表达存在偏倚。10例牙周炎患者中有3例显示1或2例明显高于自体血。3例患者中2例Vbeta6.7过表达，1例Vbeta5.2-3、Vbeta8、Vbeta12过表达。在细胞因子方面，产生IL-4和IL-6的T细胞在牙周炎中的比例高于牙龈炎。此外，il -4生成细胞的比例与B细胞：T细胞比例显著正相关。这些数据表明，牙龈T细胞不是随机从外周血中动员的，局部事件在T细胞募集或T细胞扩增水平上影响TCR库，这些T细胞的功能亚群是Th2型。少

In order to characterize the T cells infiltrating gingival tissues with periodontitis, T cell receptor repertoire and cytokine profile were investigated by means of immunohistological procedure. Periodontitis tissues were obtained at the time of periodontal surgery after completion of initial therapy. Gingivitis tissues as a control were also obtained from non-periodontitis lesions. Serial cryostat sections from each specimen were immunostained for TCR V gene products (Valpha2, Vbeta5.2-3, Vbeta5.3, Vbeta5.1, Vbeta6.7, Vbeta8, Vbeta12) and cytokines (IL-2, IFN-gamma, IL-4, IL-6) with alkaline phosphataseanti alkaline phosphatase method. Relative percentages were caluclated by dividing the number of each positive cell by total number of CD3 positive cells. Moreover, for comparison between gingival tissues and circulating pool, peripheral blood mononuclear cells from both groups were analyzed for the TCR repertoire by flow cytometry. The pattern of each TCR V gene product expression in i … More nflamed gingiva exhibited individual variation, nevertheless a consistent pattern emerged. The Vbeta5 subfamily and Vbeta6.7 were frequently used repertoires in gingiva, whereas the Valpha2 and Vbeta8 subfamily were underexpressed in most cases. Furthermore, the TCR V gene product expression in gingival tissue was biased compared with autologous peripheral blood. Three out of 10 periodontitis subjects showed 1 or 2 strikingly overrepresented repertoire comparatively with autologous blood. In these 3 subjects Vbeta6.7 was overexpressed in two cases and Vbeta5.2-3, Vbeta8 and Vbeta12 were overexpressed in one case. With respect to the cytokine profile, the proportion of IL-4- and IL-6- producing T cells was higher in periodontitis than in gingivitis. Moreover, a siginificant positive correlation between the proportion of IL-4-producing cells and the B cell : T cell ratio was noted. These data suggest that gingival T cells are not randomly mobilized from peripheral blood and that local events influence the TCR repertoire at the level of T cell recruitment or T cell expansion and the functional subsets of those T cells are Th2 type. Less