强直性脊柱炎外周血Th17细胞的检测及意义

Expression and significance of IL-17 in CD4^+ T cells in peripheral blood in patients with ankylosing spondylitis

目的探讨Th17细胞在强直性脊柱炎(AS)患者外周血中的水平及意义。方法取40例AS患者(分AS病情稳定组、活动组各20例)和健康人外周血单个核细胞(PBMC),免疫磁珠阴选CD4+T细胞,用或不用非特异性刺激剂(A-CD3、A-CD28),然后加PMA/Ion,经固定/透膜处理进行细胞内染色,流式细胞术(FCM)检测CD4+T细胞内白细胞介素17(IL-17+)/γ干扰素(IFN-γ+)、IL-17+/IL-6+水平。结果免疫磁珠阴选CD4+T细胞纯度达90%以上。AS病情活动组IL-17表达水平较病情稳定组和健康对照组显著增高(P<0.01)。用A-CD3、A-CD28和IL-23刺激后,CD4+T细胞IL-17胞内表达水平较无刺激有一定的增加。AS患者CD4+T细胞IFN-γ胞内表达水平呈现与IL-17表达相似的特点。AS患者IL-17胞内表达水平与IFN-γ和IL-6无显著相关。结论AS患者外周血CD4+T细胞胞内IL-17和IFN-γ高表达,提示分泌IL-17的Th17细胞和分泌IFN-γ的Th1细胞共同参与了AS发病过程。

Objective To study the expression and significance of Th17 cells in peripheral blood in patients with ankylosing spondylitis（AS）.Methods The venous blood specimens of peripheral blood mononuclear cell（PBMNC） were collected from 40 patients with AS and 15 healthy persons.Intracellular flow cytometry detection of IL-17/IFNγ and IL-17/IL-6 was established using Anti-CD3/Anti-CD28/IL-23 as stimulators after isolation of untouched human CD4＋T cells from PBMNC.Results The isolation of untouched human CD4＋T cells from PBMNC was effective and its purity was over 90%.The percentage of intracellular IL-17 in CD4＋T cells from AS patients was increased significantly.Such percentage in AS active group was higher than that in AS stable group and control group（P0.01）.Under Anti-CD3/Anti-CD28/IL-23 stimulation,the percentage of intracellular IL-17 was increased as well.The percentage of intracellular IFN-γ was similar to that of IL-17.There was no correlation between IL-17 and IFN-γ or IL-6.Conclusion There is an abnormal expression of IL-17 and IFN-γ in human CD4＋T cells from AS patients,suggesting that Th17 cells together with Th1 cells participates the pathogenesis of AS.