Norouzi Mojtaba, Pirestani Majid, Arefian Ehsan, Dalimi Abdolhossein, Sadraei Javid, Mirjalali Hamed
Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Department of Microbiology, School of Biology, College of Science, University of Tehran, Tehran, Iran.
Front Med (Lausanne). 2022 Aug 11;9:940332. doi: 10.3389/fmed.2022.940332. eCollection 2022.
sp. is a common intestinal parasite, possibly responsible for diarrhea, vomiting and nausea, abdominal pain, and irritable bowel syndrome. However, many studies focused on this issue due to the uncertainty of its pathogenic potential. The extracellular vesicles (EVs) are significant mediators for cellular communication, carrying biological molecules such as proteins, lipids, and nucleic acids. Compared with other parasites, little is known about the EVs. Hence the present investigation was done.
The parasites were cultured in the DMEM medium, and a 550-585 bp fragment was amplified using PCR, and sequencing was done. A commercial kit was used for exosome extraction and dynamic light scattering (DLS), flow cytometry (CD63, CD81 markers), and electron microscopy tests to determine their morphology. The human leukemia monocytic cell line (THP-1) was exposed to EVs. Next, the expression of proinflammatory and anti-inflammatory cytokines, including IL-4, IL-6, IL-10, and tumor necrosis factor-alpha (TNF-α), were measured using quantitative PCR.
Exosomes were extracted from ST1-3 sp. According to the DLS assay, the size of the exosomes was in the range of 30-100 nm. Electron microscopy images and CD63 and CD81 markers also confirmed the exosome's size, structure, and morphology. According to real-time PCR results, ST1-derived exosomes caused IL-6 and TNF-α upregulation and IL-10 and IL-4 downregulation, ST2- and ST3-derived exosomes downregulated IL-10, and ST3-derived exosomes caused IL-6 upregulation. There is a statistically significant difference ( ≤ 0.05).
To our knowledge, this is the first report of the release of exosome-like vesicles by the human parasite, , and the provided information demonstrates the role of this parasite, particularly ST1 on proinflammatory and anti-inflammatory cytokines and navigating the host response.
[寄生虫名称]是一种常见的肠道寄生虫，可能导致腹泻、呕吐和恶心、腹痛以及肠易激综合征。然而，由于其致病潜力的不确定性，许多研究都聚焦于这个问题。细胞外囊泡（EVs）是细胞间通讯的重要介质，携带蛋白质、脂质和核酸等生物分子。与其他寄生虫相比，人们对EVs了解甚少。因此开展了本研究。
将寄生虫培养于DMEM培养基中，使用PCR扩增一段550 - 585 bp的片段并进行测序。使用商业试剂盒进行外泌体提取，并通过动态光散射（DLS）、流式细胞术（CD63、CD81标记物）和电子显微镜测试来确定其形态。将人白血病单核细胞系（THP - 1）暴露于EVs。接下来，使用定量PCR测量促炎和抗炎细胞因子的表达，包括白细胞介素 - 4（IL - 4）、白细胞介素 - 6（IL - 6）、白细胞介素 - 10（IL - 10）和肿瘤坏死因子 - α（TNF - α）。
从[寄生虫名称]ST1 - 3中提取了外泌体。根据DLS测定，外泌体大小在30 - 100 nm范围内。电子显微镜图像以及CD63和CD81标记物也证实了外泌体的大小、结构和形态。根据实时PCR结果，ST1来源的外泌体导致IL - 6和TNF - α上调以及IL - 10和IL - 4下调，ST2和ST3来源的外泌体下调IL - 10，ST3来源的外泌体导致IL - 6上调。存在统计学显著差异（≤0.05）。
据我们所知，这是关于人类寄生虫[寄生虫名称]释放类似外泌体囊泡的首次报道，所提供的信息证明了这种寄生虫的作用，特别是ST1对促炎和抗炎细胞因子以及调控宿主反应的作用。
