Tang Jie, Chen Ce, Zha Cheng, Wang Zhaohua, Zhang Chen, Zeng Linli, Li Baiqing
Department of Immunology, Bengbu Medical College; Anhui Key Laboratory of Infection and Immunity, Bengbu Medical College; Science Research Centre, Bengbu Medical College, Bengbu 233030, China.
Department of Immunology, Bengbu Medical College; Anhui Key Laboratory of Infection and Immunity, Bengbu Medical College; Department of Infectious Disease, PLA 123 Hospital, Bengbu 233030, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2016 Nov;32(11):1527-1531.
Objective To investigate the differences of proportions of tumor necrosis factor α (TNF-α)-producing cells in peripheral blood γδ T cells stimulated with Mycobacterium tuberculosis heat resistant antigen (MTB-HAg) among patients with pulmonary tuberculosis (PTB), latent tuberculosis infection (LTBI) and healthy subjects (HC). Methods The peripheral blood specimens were collected from 15 normal adults, which were divided into HC group (n=9) and LTBI group (n=6), by enzyme-linked immunospot (ELISPOT) kit for diagnosis of Mycobacterium tuberculosis infection, and 12 patients with active PTB. The peripheral blood mononuclear cells (PBMCs) were separated by density gradient centrifugation and simulated with MTB-HAg for 20 hours. Then the cells were collected, and the proportions of TNF-α-producing cells in TCRαβ T cells, TCRγδ T cells, CD4αβ T cells, CD8αβ T cells, and TCR-Vδ2 T cells were measured with flow cytometry. Results The proportion of TNF-α-producing cells in γδ T cells in patients with PTB was obviously lower than that in LTBI group and HC group; the proportion of TNF-α-producing cells in Vδ2 T cells in PTB patients was apparently lower than that in LTBI and HC; the proportion of Vδ2 T cells in TNF-αγδ T cells in the peripheral blood of PTB patients was remarkably lower than that in LTBI and HC groups. The proportions of TNF-α-producing cells in peripheral αβ T cells, CD4 and CD8αβ T cells were dramatically lower than those in γδ T cells of the three according groups. Moreover, there were no statistical differences in regard with the proportions of TNF-α-producing cells in αβ T cells, and CD4 and CD8αβ T cells among the three groups. Conclusion The TNF-α production capacity of MTB-HAg specific γδ T cells and Vδ2 T cell subsets in patients with tuberculosis is obviously lower than that of LTBI and HC.
目的 探讨肺结核(PTB)患者、潜伏性结核感染(LTBI)者及健康对照(HC)外周血γδ T细胞经结核分枝杆菌耐热抗原(MTB-HAg)刺激后产生肿瘤坏死因子α(TNF-α)细胞比例的差异。方法 收集15例正常成年人外周血标本,采用酶联免疫斑点法(ELISPOT)试剂盒诊断结核分枝杆菌感染,分为HC组(n = 9)和LTBI组(n = 6),以及12例活动性PTB患者。通过密度梯度离心法分离外周血单个核细胞(PBMC),并用MTB-HAg刺激20小时。然后收集细胞,采用流式细胞术检测TCRαβ T细胞、TCRγδ T细胞、CD4αβ T细胞、CD8αβ T细胞和TCR-Vδ2 T细胞中产生TNF-α细胞的比例。结果 PTB患者γδ T细胞中产生TNF-α细胞的比例明显低于LTBI组和HC组;PTB患者Vδ2 T细胞中产生TNF-α细胞的比例明显低于LTBI组和HC组;PTB患者外周血TNF-αγδ T细胞中Vδ2 T细胞的比例明显低于LTBI组和HC组。外周αβ T细胞、CD4和CD8αβ T细胞中产生TNF-α细胞的比例明显低于三组相应γδ T细胞中的比例。此外,三组间αβ T细胞、CD4和CD8αβ T细胞中产生TNF-α细胞的比例无统计学差异。结论 结核病患者中MTB-HAg特异性γδ T细胞和Vδ2 T细胞亚群产生TNF-α的能力明显低于LTBI者和HC。