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, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran.
BMC Pulm Med. 2022 Feb 9;22(1):57. doi: 10.1186/s12890-022-01839-w.
Extracellular vesicles (EVs) play a key role in cell communication and the pathogenesis of some diseases. EVs may accelerate cell death during the course of mycobacterial infection and are also considered as a new vaccine design, drug delivery, and biomarker candidates. The current study evaluates the effects of EVs from serum samples of mycobacteria-infected patients on THP-1 monocytes and PBMC cells.
EVs were purified from the serum, then cultured separately with THP-1 monocytes and PBMCs. The cell death was determined through annexin V-FITC and PI staining. GW4869, an EVs inhibitor, was used to determine if EVs released from serum could increase THP-1 monocytes cell death.
The cell death was significantly increased in the presence of 10 µg/ml and 5 µg/ml concentrations of the purified EVs (p < 0.05). Minimal cell death was determined in 2.5 µg/ml and 1.2 µg/ml (p < 0.05). Up to 85% of the cells were viable in the presence of the GW4869 inhibitor (p < 0.05).
Direct infection of the cells with EVs released from mycobacteria-infected patients samples, the multiplicity of infection with the EVs, and virulent or avirulent mycobacteria may change the status of the cell death. The isolated EVs from serum samples of patients with mycobacterial infection accelerated cell death, which means that they might not be considered as an optimal tool for developing drug delivery and vaccine against tuberculosis.
细胞外囊泡 (EVs) 在细胞通讯和某些疾病的发病机制中起着关键作用。EVs 可能会在分枝杆菌感染过程中加速细胞死亡，并且也被认为是新的疫苗设计、药物输送和生物标志物候选物。本研究评估了分枝杆菌感染患者血清样本中的 EVs 对 THP-1 单核细胞和 PBMC 细胞的影响。
从血清中纯化 EVs，然后分别与 THP-1 单核细胞和 PBMC 培养。通过 Annexin V-FITC 和 PI 染色来确定细胞死亡。使用 GW4869（EVs 抑制剂）来确定从血清中释放的 EVs 是否会增加 THP-1 单核细胞的细胞死亡。
在存在 10 µg/ml 和 5 µg/ml 浓度的纯化 EVs 时，细胞死亡明显增加（p < 0.05）。在 2.5 µg/ml 和 1.2 µg/ml 时确定了最小的细胞死亡（p < 0.05）。在存在 GW4869 抑制剂的情况下，多达 85%的细胞存活（p < 0.05）。
直接用分枝杆菌感染患者样本中释放的 EVs 感染细胞，EVs 的感染复数以及毒力或非毒力分枝杆菌可能会改变细胞死亡的状态。从分枝杆菌感染患者血清样本中分离的 EVs 加速了细胞死亡，这意味着它们可能不被认为是开发针对结核病的药物输送和疫苗的理想工具。
