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
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...
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| Published in | 南方医科大学学报 Vol. 37; no. 11; pp. 1442 - 1447 |
|---|---|
| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| Published |
解放军123医院传染科,安徽 蚌埠 233010%蚌埠医学院科研中心,安徽 蚌埠,233030%安徽省感染与免疫重点实验室(蚌埠医学院),安徽 蚌埠,233030%蚌埠市传染病医院呼吸科,安徽 蚌埠,233010%蚌埠医学院免疫学教研室,安徽 蚌埠233030
2017
蚌埠医学院科研中心,安徽 蚌埠233030%蚌埠医学院免疫学教研室,安徽 蚌埠233030 蚌埠医学院免疫学教研室,安徽 蚌埠233030 安徽省感染与免疫重点实验室(蚌埠医学院),安徽 蚌埠233030 |
| Subjects | |
| Online Access | Get full text |
| ISSN | 1673-4254 |
| DOI | 10.3969/j.issn.1673-4254.2017.11.03 |
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| Abstract | 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(P<0.01);与PHA刺激组相比,γδT细胞中TNF-α和IFN-γ产生细胞比例增高(P<0.01),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(P<0.01);肺结核患者IFN-γ产生细胞数量低于潜伏性结核感染者(P<0.01)。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。 |
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| AbstractList | 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(P<0.01);与PHA刺激组相比,γδT细胞中TNF-α和IFN-γ产生细胞比例增高(P<0.01),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(P<0.01);肺结核患者IFN-γ产生细胞数量低于潜伏性结核感染者(P<0.01)。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。 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(P<0.01);与PHA刺激组相比,γδT细胞中TNF-α和IFN-γ产生细胞比例增高(P<0.01),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(P<0.01);肺结核患者IFN-γ产生细胞数量低于潜伏性结核感染者(P<0.01).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_FL | Objective To investigate the effects of low molecular peptide of Mycobacterium tuberculosis heat-resistant antigen (Mtb-HAg-10k) on the production of tumor necrosis factor-α(TNF-α) and interferon-γ(IFN-γ) in peripheral blood T cells and test the feasibility of differential diagnosis between pulmonary tuberculosis (PTB) and latent tuberculosis infection (LTBI) by assessing the number of Mtb-HAg-10k-stimulated IFN-γ-producing T cells. Methods Peripheral blood mononuclear cells (PBMCs) were separated from the peripheral blood of 10 healthy adults, 6 individuals with LTBI and 13 patients with PTB. The PBMCs were cultured in the presence of Mtb-HAg-10k obtained by ultrafiltration centrifugation, with Mtb-HAg and phytohaemagglutinin (PHA) as the controls. The proportions of TNF-α-and IFN-γ-producing cells in the T cell subsets were detected by flow cytometry (FCM), and the number of IFN-γ-producing cells from patients with PTB and LTBI was detected with ELISPOT. Results Flow cytometry showed that Mtb-HAg-10k exposure resulted in a significantly higher proportion of TNF-α-producingγδT cells than that of IFN-γ-producingγδT cells in the PBMCs (P<0.01). Compared with the PBMCs exposed to PHA, the PBMCs exposed to Mtb-HAg-10k exhibited a significantly greater proportion of γδT cells that produced both TNF-α and IFN-γ (P<0.01) but a significantly lower proportion of αβT cells producing both TNF-α and IFN-γ (P<0.01). Mtb-HAg-10k exposure of the PBMCs caused a significant reduction in the number of IFN-γ-producing cells as compared with Mtb-HAg and PHA treatments (P<0.01), and this reduction was more obvious in PBMCs from patients with PTB than in those from individuals with LTBI (P<0.01). Conclusion Mtb-HAg-10k can markedly induce γδT cells in the PBMCs to produce TNF-α and IFN-γ, and detection of the number of IFN-γ-producing cells in the PBMCs following Mtb-HAg-10k stimulation helps in the differential diagnosis between pulmonary tuberculosis and latent tuberculosis infection. |
| Author | 唐洁 陈策 查成 常见荣 方强 王兆华 李柏青 |
| AuthorAffiliation | 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 |
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| Author_FL | ZHA Chen TANG Jie CHANG Jianrong CHEN Ce FANG Qiang LI Baiqing WANG Zhaohua |
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| Keywords | pulmonary tuberculosis Mycobacterium tuberculosis heat resistant antigen interferon-γ 干扰素-γ 结核杆菌耐热抗原 tumor necrosis factor-α latent tuberculosis infection γδT细胞 肿瘤坏死因子-α 肺结核 潜伏性结核感染 γδT cells |
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| Snippet | 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... 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... |
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| SubjectTerms | 结核杆菌耐热抗原;γδT细胞;肿瘤坏死因子-α;干扰素-γ;肺结核;潜伏性结核感染 |
| Title | 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 |
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