Department of Parasitology, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan.
Department of Medical Laboratory Science, College of Medical Science and Technology, I-Shou University, Kaohsiung 824, Taiwan.
Genes (Basel). 2022 Mar 17;13(3):531. doi: 10.3390/genes13030531.
Accumulated evidence suggests that the endosymbiotic (TVV) may play a role in the pathogenesis and drug susceptibility of . Several reports have shown that extracellular vesicles (EVs) released from TVV-positive (TVV+) trichomonads can modulate the immune response in human vaginal epithelial cells and animal models. These results prompted us to examine whether EVs released from TVV+ isolates contained TVV. We isolated small extracellular vesicles (sEVs) from six isolates that were either TVV free (ATCC 50143), harbored a single (ATCC 30236, ATCC 30238, T1), two (ATCC PRA-98), or three TVV subspecies (ATCC 50148). The presence of TVV subspecies in the six isolates was observed using reverse transcription-polymerase chain reaction (RT-PCR). Transmission electron microscopy (TEM) confirmed the presence of cup-shaped sEVs with a size range from 30-150 nm. tetraspanin (TvTSP1; TVAG_019180), the classical exosome marker, was identified in all the sEV preparations. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis showed that all the sEVs isolated from TVV+ isolates contain viral capsid proteins derived from the same TVV subspecies in that isolate as demonstrated by RT-PCR. To provide more comprehensive information on the TVV subspecies population in other isolates, we investigated the distribution of TVV subspecies in twenty-four isolates by mining the New-Generation Sequencing (NGS) RNAseq datasets. Our results should be beneficial for future studies investigating the role of TVV on the pathogenicity of and the possible transmission of virus subspecies among different isolates via sEVs.
积累的证据表明，内共生体（TVV）可能在的发病机制和药物敏感性中发挥作用。有几项报告表明，从 TVV 阳性（TVV+）滴虫中释放的细胞外囊泡（EVs）可以调节人阴道上皮细胞和动物模型中的免疫反应。这些结果促使我们检查从 TVV+分离株中释放的 EV 是否含有 TVV。我们从六个分离株中分离出小细胞外囊泡（sEV），这些分离株要么不含 TVV（ATCC 50143），要么含有单个（ATCC 30236、ATCC 30238、T1）、两个（ATCC PRA-98）或三个 TVV 亚种（ATCC 50148）。使用逆转录聚合酶链反应（RT-PCR）观察六个分离株中 TVV 亚种的存在。透射电子显微镜（TEM）证实存在杯状 sEV，其大小范围为 30-150nm。tetraspanin（TvTSP1；TVAG_019180），即经典的外泌体标记物，在所有 sEV 制剂中均被鉴定出来。液相色谱-串联质谱（LC-MS/MS）分析表明，从 TVV+分离株中分离出的所有 sEV 均含有与该分离株中相同 TVV 亚种的病毒衣壳蛋白，这通过 RT-PCR 得到证实。为了提供有关其他分离株中 TVV 亚种群体的更全面信息，我们通过挖掘新一代测序（NGS）RNAseq 数据集，研究了二十四株分离株中 TVV 亚种的分布。我们的结果应该有助于未来研究 TVV 在致病性和病毒亚种通过 sEV 在不同分离株之间传播的可能性。
