Javadi Alireza, Shamaei Masoud, Tabarsi Payam, Ainy Elaheh, Kazemi Bahram
Virology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Clinical Tuberculosis and Epidemiology Research Center, NRITLD, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Tanaffos. 2022 Apr;21(4):434-447.
Extracellular vesicles (EVs) may accelerate cell death during the course of infection. Mycobacteria could invade the host's immune system and survive in the host by modulation of miRNAs. MiRNAs' differential expressions can serve as biomarkers. This study evaluates THP-1 monocyte cell death by EVs from serum of patients with mycobacteria and assesses serum-derived exosomal miRNAs to increase or decrease THP-1 monocyte cell death.
EVs were purified from serum of patients with mycobacteria and cultured with THP-1 monocyte. The cell death was determined via annexin V-FITC and PI staining. The microRNA was isolated from serum-derived EVs of the patients. Expression level of Hsa-miR-20a-5p, Hsa-miR-29a, Hsa-miR-let7e, and Hsa-miR-155 was assessed using qRT-PCR.
Cell death was accelerated in 10 and 5 μg/ml concentrations of the EVs (p<0.05). Minimum cell death was seen in 2.5 and 1.2 μg/ml concentrations (p<0.05). In tuberculosis (TB) patients, expression of miR-20a-5p, miR-29a, and miR-let7e were significantly enhanced (p≤0.0001), but miR-155 expression reduced. ROC analysis showed diagnostic biomarkers of miRNAs with an AUC=0.6933 for miR-20, AUC=0.6011 for miR-29a, AUC=0.7322 for miR-let7e, and AUC=0.7456 for miR-155 for active tuberculosis. Expression of miR-let7e, 20a, and 29a in vs. was overexpressed (P≤0.01, P≤0.0001, and P≤0.0001, respectively). Also miRs let7e and 20a expression was accelerated in vs. (P≤0.0001 and P≤0.002, respectively).
EVs accelerates cell death and may not be ideally considered for drug delivery and vaccine developments. Circulating exosomal microRNA MiR-20, miR-let7e, and miR-155 facilitate development of potential biomarkers of pulmonary tuberculosis and non-tuberculosis.
细胞外囊泡（EVs）可能在感染过程中加速细胞死亡。分枝杆菌可侵入宿主免疫系统并通过调节微小RNA（miRNAs）在宿主体内存活。miRNAs的差异表达可作为生物标志物。本研究评估来自分枝杆菌患者血清的EVs对THP-1单核细胞死亡的影响，并评估血清来源的外泌体miRNAs对THP-1单核细胞死亡的增加或减少作用。
从分枝杆菌患者血清中纯化EVs，并与THP-1单核细胞共培养。通过膜联蛋白V-异硫氰酸荧光素（annexin V-FITC）和碘化丙啶（PI）染色确定细胞死亡情况。从患者血清来源的EVs中分离微小RNA。使用定量逆转录聚合酶链反应（qRT-PCR）评估人miR-20a-5p、人miR-29a、人miR-let7e和人miR-155的表达水平。
10μg/ml和5μg/ml浓度的EVs可加速细胞死亡（p<0.05）。在2.5μg/ml和1.2μg/ml浓度下观察到最低的细胞死亡（p<0.05）。在结核病（TB）患者中，miR-20a-5p、miR-29a和miR-let7e的表达显著增强（p≤0.0001），但miR-155表达降低。受试者工作特征（ROC）分析显示，miRNAs作为诊断生物标志物，对于活动性结核病，miR-20的曲线下面积（AUC）=0.6933，miR-29a的AUC=0.6011，miR-let7e的AUC=0.7322，miR-155的AUC=0.7456。与对照组相比，miR-let7e、20a和29a在病例组中的表达上调（分别为P≤0.01、P≤0.0001和P≤0.0001）。此外，与对照组相比，miR-let7e和20a在病例组中的表达加速（分别为P≤0.0001和P≤0.002）。
EVs加速细胞死亡，可能不太适合用于药物递送和疫苗开发。循环外泌体微小RNA miR-20、miR-let7e和miR-155有助于开发肺结核和非结核的潜在生物标志物。
