【摘要】 目的通过对比分析结核病患者与健康个体外周血中恒定自然杀伤T细胞(invariant nature killer T cells,iNKT)的比例、表型以及功能,认识iNKT细胞在结核病患者体内的功能状态,为探讨基于iNKT细胞的干预措施在结核病防治中的应用奠定基础。方法收集武汉市新洲区人民医院收治的21例结核病患者及14例健康体检者外周血,分离外周血单个核细胞(peripheral blood mononuclear cell,PBMC)。对PBMC进行细胞表面染色,用流式细胞仪分别测定iNKT细胞的比例和表面分子CD69、CD62L、CD122以及CD161的表达;用丙二醇甲醚醋酸酯/离子霉素(phorbol-12-myristate-13-acetate/Ionomycin,PMA/ION)刺激PBMC,通过胞内细胞因子染色,测定iNKT细胞潜在的合成分泌IFN-γ和IL-4的能力;并用α-半乳糖神经酰胺(α-Galactosylceramide,α-GalCer)刺激培养1周,测定结核病患者iNKT细胞的反应性及扩增效率。结果相对于健康个体,结核病患者外周血中的iNKT细胞百分比明显降低;结核病患者iNKT细胞的CD69表达水平显著高于健康个体,而其CD62L、CD122及CD161的表达量与健康个体无明显差异;结核病患者和健康个体的iNKT细胞在PMA/ION刺激后分泌IFN-γ和IL-4的能力无明显差异;然而结核病患者的iNKT细胞对α-GalCer的反应能力明显减弱,表现为被α-GalCer刺激后的扩增能力低于健康个体。结论结核病患者体内iNKT细胞的数量与功能异常,提示恢复iNKT细胞数量与功能是促进iNKT细胞抗结核效应的关键因素。更多还原

【Abstract】 Objective To examine the function of invariant nature killer T cells(iNKT)in prevention and treatment of tuberculosis(TB)by comparing the proportion,phenotype and function of iNKT cells in peripheral blood of tubercular patients and healthy individuals.Methods Peripheral blood mononuclear cells(PBMC)were isolated from 21 TB patients who were admitted to the People’s Hospital of Xinzhou District and 14 healthy subjects,Wuhan.Cell surface staining of PBMC was carried out,and the ratio of iNKT cells and the expression of surface molecules CD69,CD62 L,CD122 and CD161 were determined by flow cytometry.Phorbol-12-myristate-13-acetate/ionomycin(PMA/ION)was used to stimulate PBMC and intracellular cytokine staining to determine the potential of iNKT cells to secrete IFN-γand IL-4.The reactivity and amplification efficiency of iNKT cells from TB patients were determined afterα-galactosylceramide(α-GalCer)treatment.Results The proportion of iNKT cells was significantly lower and the CD69 expression in iNKT cells was significant higher in TB patients than in healthy individuals.However,there were no differences in the expression of CD62 L,CD122 and CD161 between TB patients and healthy individuals.Similar,no difference was noted in the ability of iNKT to secret IFN-γand IL-4 after PMA/ION stimulation between TB patients and healthy individuals.Moreover,the iNKT cells showed weaker response toα-GalCer in TB patients as evidenced by the fact that the proliferation ability ofα-GalCer-treated iNKT cells in TB patients was lower than that in healthy individuals.Conclusion These results indicate that the number and function of iNKT cells in TB patients are abnormal,suggesting that restoring the number and function of iNKT cells is a key factor in promoting the anti-tuberculosis effect of iNKT cells.更多还原