基于结核杆菌耐热抗原小分子多肽刺激人外周血T细胞产生TNF-α和IFN-γ鉴别肺结核和潜伏性结核感染

发布时间：2018-04-02 09:12

本文选题：结核杆菌耐热抗原　切入点：γδT细胞　出处：《南方医科大学学报》2017年11期

【摘要】：目的研究结核杆菌耐热抗原小分子多肽(Mtb-HAg-10k)刺激人外周血T细胞产生细胞因子肿瘤坏死因子-α(TNF-α)和干扰素-γ(IFN-γ)的作用特点及肺结核患者和潜伏性感染者IFN-γ产生细胞的数量差异,初步探讨Mtb-HAg-10k作为诊断抗原鉴别肺结核和潜伏性结核感染的可行性。方法以超滤离心法获得的Mtb-HAg-10k为刺激剂作用于人外周血单个核细胞(PBMCs),并以Mtb-HAg、植物血凝素PHA作为对照,用多色流式细胞术检测T细胞亚群中TNF-α和IFN-γ产生细胞比例,酶联免疫斑点法检测肺结核患者和潜伏性感染者PBMCs中IFN-γ产生细胞数量。结果流式细胞术检测人外周血Mtb-HAg-10k特异性γδT细胞中TNF-α产生细胞比例显著高于IFN-γ产生细胞比例(P0.01);与PHA刺激组相比,γδT细胞中TNF-α和IFN-γ产生细胞比例增高(P0.01),而αβT细胞中TNF-α和IFN-γ产生细胞比例显著降低;酶联免疫斑点法检测Mtb-HAg-10k刺激的健康者PBMCs中IFN-γ产生细胞数量低于Mtb-HAg刺激组和PHA对照组(P0.01);肺结核患者IFN-γ产生细胞数量低于潜伏性结核感染者(P0.01)。结论以Mtb-HAg-10K为刺激剂可使人外周血γδT细胞优势产生TNF-α和IFN-γ,通过酶联免疫斑点法检测PBMCs中IFN-γ产生细胞数量差异有助于鉴别肺结核和潜伏性结核感染。
[Abstract]:Objective to study the effects of Mtb-HAg-10k) on the production of cytokine tumor necrosis factor- 伪 (TNF- 伪) and interferon- 纬 (IFN- 纬) in human peripheral blood T cells and the difference in the number of IFN- 纬 producing cells between pulmonary tuberculosis patients and latent infections. Methods Mtb-HAg-10k obtained by ultrafiltration centrifugation was used as stimulant to treat human peripheral blood mononuclear cells (PBMC), and Mtb-HAg and phytohemagglutinin (PHA) were used as control. The percentage of TNF- 伪 and IFN- 纬 producing cells in T cell subsets was detected by polychromatic flow cytometry. The number of IFN- 纬 producing cells in PBMCs of pulmonary tuberculosis and latent infection was detected by enzyme-linked immunoblot assay. Results the percentage of TNF- 伪 producing cells in Mtb-HAg-10k specific 纬未 T cells in peripheral blood by flow cytometry was significantly higher than that in IFN- 纬 producing cells. The ratio of TNF- 伪 and IFN- 纬 producing cells in 纬未 T cells was higher than that in PHA stimulation group, while the proportion of TNF- 伪 and IFN- 纬 producing cells in 伪尾 T cells decreased significantly. The number of IFN- 纬 producing cells in PBMCs of Mtb-HAg-10k stimulated healthy persons was lower than that of Mtb-HAg stimulation group and PHA control group, and the number of IFN- 纬 producing cells in pulmonary tuberculosis patients was lower than that in latent TB infected persons. Conclusion Mtb-HAg-10K can be used as a stimulant to increase the number of IFN- 纬 producing cells. Human peripheral blood 纬未 T cells produce TNF- 伪 and IFN- 纬 preferentially. Detection of the difference in the number of IFN- 纬 producing cells in PBMCs by enzyme-linked immunoblot assay is helpful to distinguish tuberculosis from latent tuberculosis infection.
【作者单位】：蚌埠医学院免疫学教研室;安徽省感染与免疫重点实验室(蚌埠医学院);蚌埠医学院科研中心;解放军123医院传染科;蚌埠市传染病医院呼吸科;
【基金】：安徽高校科研平台创新团队项目(2016-40) 安徽高校省级科研项目(KJ2011Z248) 蚌埠医学院科技发展基金(BYKF1402)
【分类号】：R521

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