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Title: [Dynamic analysis of tuberculosis specific mononuclear cells in peripheral blood of HIV patients with tuberculosis].
Author: Zou S, Zhu Q, Guo YN, Xiang PZ, Mo GL, Chen JH, Liu K, Liang K.
Journal: Zhonghua Jie He He Hu Xi Za Zhi; 2022 Nov 12; 45(11):1109-1116. PubMed ID: 36344228.
Abstract:
Objective: To evaluate the response of peripheral blood mononuclear cells (PBMCs) in patients with human immunodeficiency virus (HIV) combined with active tuberculosis (TB) to TB-specific antigen stimulation. Methods: From January to December, 2018, individuals infected with both HIV and TB (HIV/TB group) were taken as the study subjects. Individuals infected with HIV alone (HIV group), individuals infected with TB alone (TB group) and healthy people (Health control group, HC group) were taken as the control groups. PBMCs were isolated and stimulated with purified protein derivative of bacillus calmette-guerin (BCG-PPD). The expression of surface molecules in T cells (CD3⁺, CD4⁺, CD8⁺) and monocytes (CD14⁺) and the percentages of Interferon (IFN)-γ and tumor necrosis factor (TNF)-α were detected by cell surface molecular staining, direct intracellular cytokine staining and flow cytometry (CD3^- lymphocytes were mainly B lymphocytes and NK cells). Analysis of non-parametric data was used to compare the data between the two groups, and paired t-test was used to compare the data before and after PPD stimulation in each group. Results: Before PPD stimulation, the percentage of IFN-γ⁺ CD8⁺ cells in the peripheral blood of HIV/TB group(mean 0.52%) was significantly lower than that in TB group(mean 0.94%, P=0.010). The TNF-α⁺cell percentages in CD3⁺, CD4⁺, CD8⁺, or CD14⁺ cells in the HIV/TB group(mean 19.2%) were significantly lower than those in the HIV group(mean 31.9%, P=0.002). The percentage of TNF-α secreted by monocytes in the HIV group was significantly lower than that in the HC group. The percentages of IFN-γ⁺ CD8⁺ and IFN-γ⁺ CD3^- cells in the peripheral blood of the TB group (mean 0.94%) were significantly higher than thoset in the HC group(mean 0.51%, P=0.020), while the percentages of TNF-α⁺ cells in each subsets of PBMCs were significantly lower than those in the HC group. After PPD stimulation, the percentage of IFN-γ⁺ CD8⁺ cells in the HIV/TB group was significantly lower than that in the TB group(P=0.008), and the change was more marked than that before stimulation. The percentage of IFN-γ⁺ CD8⁺ cells in the HIV group(mean 0.20%) was lower than that in the HC group (mean 0.52%, P=0.044). The percentage of IFN-γ⁺ CD3^- in the TB group was significantly higher than in the HC group. There were no significant differences in TNF-α⁺ cell percentages in the 3 groups compared with the control group after PPD stimulation. The percentages of IFN-γ⁺ CD4⁺ cells in the HC and the TB groups were significantly increased after PPD stimulation in each group (P=0.002, P=0.001, respectively). However, there were no significant differences of IFN-γ⁺ CD4⁺ cell percentages in the HIV/TB group and the HIV group. The percentages of TNF-α production by monocytes were significantly increased after PPD stimulation in all groups. Conclusions: Chronic Mycobacterium tuberculosis (MTB) infection reduced the ability of PBMCs to produce TNF-α. For patients with TB infection, the production of TNF-α was reduced when combined with HIV infection. The capacity of CD8⁺ and CD3^- lymphocytes to produce IFN-γ was increased in TB patients, while the capacity of CD8⁺ T cells to produce IFN-γ was decreased with co-infection of HIV. Infection of HIV weakened the immune response to MTB infection, which made the clinical diagnosis and treatment of TB more difficult. 目的： 探讨人类免疫缺陷病毒（HIV）合并结核病（TB）患者外周血单个核细胞（PBMC）分泌细胞因子及对结核特异性抗原刺激的反应。 方法： 2018年1—12月，以HIV合并TB患者（HIV/TB组）为研究对象，以单纯HIV感染者（HIV组），单纯TB患者（TB组）及健康人群（HC组）为对照组。分离PBMC并用卡介菌纯蛋白衍化物（BCG-PPD）刺激。通过细胞表面分子染色、直接细胞内细胞因子染色及流式细胞术检测这些人群PBMC中分泌干扰素-γ（interferon-γ，IFN-γ）和肿瘤坏死因子-α（TNF-α）的淋巴细胞（CD3⁺、CD3^-、CD4⁺、CD8⁺）及单核细胞（CD14⁺）的比例（其中CD3^-淋巴细胞主要为B淋巴细胞及NK细胞）。两组间数据比较采用非参数检验，每组研究对象PPD刺激前后数据的比较采用配对t检验。 结果： 未经PPD刺激时，HIV/TB组（均值0.52%）IFN-γ⁺CD8⁺/CD8⁺百分比显著低于TB组（均值0.94%，P=0.010），分泌TNF-α的PBMC各亚群细胞百分比均显著低于HIV组；HIV组（均值19.2%）TNF-α⁺CD14⁺/CD14⁺百分比显著低于HC组（均值31.9%，P=0.002）；TB组（均值 0.94%）IFN-γ⁺CD8⁺/CD8⁺百分比显著高于HC组（均值0.51%，P=0.020），而分泌TNF-α的PBMC各亚群细胞百分比显著低于HC组。PPD刺激后，HIV/TB组（均值0.20%）IFN-γ⁺CD8⁺/CD8⁺百分比明显低于TB组（均值0.56%，P=0.008），且比刺激前差异更显著；HIV组（均值0.24%）IFN-γ⁺CD8⁺/CD8⁺百分比较HC组降低（均值0.52%，P=0.044）。PPD刺激后，各组间分泌TNF-α的PBMC各亚群细胞百分比与HC组差异无统计学意义。各组组内PPD刺激前后相比，HC组及TB组IFN-γ⁺CD4⁺/CD4⁺百分比显著增加，而HIV/TB组及HIV组差异无统计学意义，四组TNF-α⁺CD14⁺/CD14⁺百分比均明显增加。 结论： 慢性结核分枝杆菌（MTB）感染使PBMC产生TNF-α的能力下降，而合并HIV感染加重该趋势。HIV感染时单核细胞产生TNF-α的能力下降。TB患者的CD8⁺及CD3^-淋巴细胞产生IFN-γ的能力增强，而合并HIV感染时IFN-γ的产生减少主要来自CD8⁺T细胞。HIV感染使机体对MTB感染的免疫反应减弱，从而给TB的临床诊断及治疗带来更多困难。.

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