AlShwaimi Emad, Purcell Patricia, Kawai Toshihisa, Sasaki Hajime, Oukka Mohamed, Campos-Neto Antonio, Stashenko Philip
Department of Restorative Dentistry and Biomaterials Sciences, Harvard School of Dental Medicine, Boston, MA, USA.
J Endod. 2009 Sep;35(9):1229-33. doi: 10.1016/j.joen.2009.06.006.
T-regulatory (Treg, CD4+ FOXP3+) cells constitute a unique subpopulation of CD4+ T cells that inhibit T-cell responses and prevent disease development/exacerbation in models of autoimmunity. In the present study, we tested the hypothesis that Treg cells are induced in periapical lesions by dental pulp infection.
In situ hybridization (ISH) was used to localize FOXP3+ cells on day 21 after pulp exposure of the first molar teeth and infection with bacteria from the oral environment. FOXP3/GFP knock-in transgenic mice were used to quantify FOXP3+ Treg cells that infiltrate into periapical lesions by flow cytometry on days 7, 14, and 21 after infection. Periodontal ligament from uninfected teeth served as a negative control.
ISH showed strong signals that showed the presence of FOXP3+ cells mainly at the periphery of periapical lesions. In contrast, no positive cells were present in the periodontal ligament of uninfected controls. Flow cytometry showed an increase in the number of FOXP3+ Treg beginning between day 7 and day 14 (0.69% of the infiltrate) after infection and increased to day 21 (0.94%) (p < 0.05 and p < 0.001, respectively, vs uninfected controls). Treg were also increased in number in draining cervical lymph nodes after pulpal infection.
These results show that Treg cells are induced to infiltrate into periapical lesions by pulpal infection and suggest that they increase in a time-dependent manner.
调节性T(Treg,CD4+FOXP3+)细胞构成CD4+T细胞的一个独特亚群,可抑制T细胞反应,并在自身免疫模型中预防疾病发展/加重。在本研究中,我们检验了牙髓感染可在根尖周病变中诱导Treg细胞产生这一假说。
在第一磨牙牙髓暴露并感染口腔环境中的细菌后第21天,采用原位杂交(ISH)定位FOXP3+细胞。利用FOXP3/GFP基因敲入转基因小鼠,通过流式细胞术对感染后第7天、14天和21天浸润到根尖周病变中的FOXP3+Treg细胞进行定量分析。未感染牙齿的牙周韧带作为阴性对照。
ISH显示强信号,表明FOXP3+细胞主要存在于根尖周病变的周边。相比之下,未感染对照的牙周韧带中没有阳性细胞。流式细胞术显示,感染后第7天至第14天之间,FOXP3+Treg细胞数量开始增加(占浸润细胞的0.69%),并在第21天增加至0.94%(与未感染对照相比,p分别<0.05和<0.001)。牙髓感染后,引流颈部淋巴结中的Treg数量也增加。
这些结果表明,牙髓感染可诱导Treg细胞浸润到根尖周病变中,并提示它们以时间依赖性方式增加。