炎性牙周膜干细胞影响T细胞迁移及活化的初步研究

Inflammatory PDLSCs promotes T cell migration and activation:A pilot study

目的:初步探讨炎性牙周膜干细胞(periodontalligamentstemcells,PDLSCs)在牙周炎中对T细胞免疫的影响及可能机制。方法:使用磷酸盐缓冲液(phosphate buffered saline,PBS)和脂多糖(lipopolysaccharide,LPS)刺激PDLSCs模拟健康及炎性环境下的PDLSCs,收集2组PDLSCs条件培养液(conditioned medium,CM),即健康PDLSCs条件培养液(CON-CM组)和炎性PDLSCs条件培养液(LPS-CM组),并收集同批次培养液为阴性对照(NC组),分别将其作用于人正常T淋巴细胞系H9细胞,观察不同CM对T细胞增殖和迁移能力的作用。进一步采用植物血凝素-L(phytohemagglutinin-L,PHA-L)刺激T细胞免疫活化,采用CM作用于活化状态下的H9细胞,并检测T细胞免疫活化相关基因白细胞介素-2(interleukin-2,IL-2)、γ干扰素(interferon-γ,IFN-γ)、白细胞介素-17(interleukin-17,IL-17)、CC趋化因子受体6(CC chemokine receptor 6,CCR6)表达的变化,同时观察LPS刺激对PDLSCs分泌CC趋化因子配体20(CC chemokine ligand 20,CCL20)的影响。结果:与NC组和CON-CM组相比,LPS-CM组能促进T细胞的增殖、迁移,及IL-2、IFN-γ、IL-17和CCR6的表达(P<0.05),且在LPS的刺激下,PDLSCs分泌的CCL20含量明显增多(P<0.001)。结论:正常PDLSCs具有一定免疫调节作用,而炎性PDLSCs可能通过CCL20-CCR6轴促进了T细胞的增殖、迁移和活化等免疫生物学行为。

Objective:To explore the inflammatory periodontal ligament stem cells(PDLSCs)on T cell immunity in periodontitis conditions and its possible mechanism.Methods:PDLSCs were stimulated with phosphate buffered saline(PBS)and lipopolysaccharide(LPS)to mimic PDLSCs in healthy and inflammatory environments,and then the two groups of conditioned medium(CM)were collected,namely healthy PDLSCs CM(CON-CM group)and inflammatory PDLSCs CM(LPS-CM group).The same batch of culture medium was collected as the negative control(NC group)as well.They were used to act on human normal T lymphocyte line H9 cells respectively to observe the effects of different CM on T cell proliferation and migration.Furthermore,phytohemagglutinin-L(PHA-L)was used to stimulate T cell immune activation,and CM was used to treat H9 cells in the activated state,then the expression of T cell immune activation related genes—interleukin-2(IL-2),interferon-γ(IFN-γ),interleukin-17(IL-17)and CC chemokine receptor 6(CCR6)were detected,and the effect of LPS stimulation on the secretion of CC chemokine ligand 20(CCL20)by PDLSCs was observed.Results:Compared with the NC group and the CON-CM group,the LPS-CM group could promote the proliferation and migration of T cells,as well as the expression of IL-2,IFN-γ,IL-17 and CCR6 mRNA(P<0.05).While the content of CCL20 secreted by PDLSCs was significantly increased under LPS stimulation(P<0.001).Conclusion:This study provides evidence that normal PDLSCs have certain immunomodulatory effects,while inflammatory PDLSCs may promote T cell proliferation,migration,activation and other immunobiological behaviors through the CCL20-CCR6 axis.