结核病患者外周血iNKT细胞数量与功能缺陷  

Number and Function Defects of iNKT Cells in Peripheral Blood of Tuberculosis Patients

作　　者：冯显红 谭晓晟 周平 翁秀芳[2] Feng Xianhong;Tan Xiaosheng;Zhou Ping(Clinical Laboratory,People’s Hospital of Wuhan Xinzhou District;Department of Immunology,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]武汉市新洲区人民医院检验科,武汉431400 [2]华中科技大学同济医学院基础医学院免疫学系,武汉430030

出　　处：《华中科技大学学报（医学版）》2018年第5期533-538,567,共7页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：武汉市卫生计生委科研基金资助项目(No.WX17C28);国家自然科学基金资助项目(No.31570913)

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。Objective To examine the function of invariant nature killer T ccllsC iNKT.)in prevention and treatment of tuberculosis(T'B)by comparing the proportion,phenotype and function of iNKT cells in peripheral blood of tubercular patients and healthy individuals.Methods Peripheral blood mononuclear cclls(PBMC)were isolated from 21 T'B patients who were ad mitted to the People7s 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)CD62L?CD122 and CD161 were determined by flow cytometry.Phorbol-12-myristatc-13-acctatc/ionomycin(PMA/](.)N)was used to stimulate PBMC and intracellular cytokine staining to determine the potential of iNKT cells to secrete IF^N-y and IL-4.The reactivity and amplification efficiency of iNKT cells from T'B patients were determined after a-galactosylccramidc(a-GalCcr)treatmem.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,therewere no differences in the expression of CD62L,CD122 and CD161 betweenTB patients and healthy individuals Similar,no diffcrcncewas noted in the ability of iNKT to secret FN-YandIL-4aftcrPMA/IONstimulationbe-tweenTB patients and healthy individuals Moreover,the iNKT cells showedweaker response to a-GalCcr inTB patients as evi-dcnccd by the fact that the proliferation ability of a-GalCcr-trcatcd iNKT cells in T'B patients was lower than that in healthy in-dividualt.Conclusion T'hcse results indicate that the number and function of iNKT cells in T'B 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 cell.

关 键 词：结核病 INKT细胞 表型 功能 Α-GALCER 

分 类 号：R392[医药卫生—免疫学]