冠状病毒(CoV)已成为一种公共卫生危机,在人类和某些动物中引起许多疾病。研究已经确定了小的,称为细胞外囊泡(EV)的脂质结合结构作为病毒可以进入宿主细胞的机制,传播,逃避宿主的免疫防御.电动汽车能够包装和携带多种病毒化合物,包括蛋白质,遗传物质,脂质,和受体蛋白。我们提出冠状病毒可以改变电动汽车的产量和含量,以及影响EV在宿主细胞中的生物发生和组成。
■在当前的研究中,Crandell-Rees猫肾(CRFK)细胞在48h和72h时间点以2,500个感染单位(IFU)的感染复数(MOI)在无外泌体培养基中感染猫冠状病毒(FCoV)。分析细胞活力,发现由于FCoV感染显著降低9%(48小时)和15%(72小时)。电动汽车通过超速离心分离,并通过扫描电子显微镜(SEM)分析了分离的电动汽车的表面形貌。
■NanoSight粒子跟踪分析(NTA)证实,对照EV的平均粒径为131.9nm和126.6nm,而FCoV感染的EV在48和72小时分别为143.4nm和120.9nm,分别。总DNA,RNA,在两个孵育时间点,在分离的EV中测定蛋白质水平;然而,总蛋白在48h时显著增加。TMPRSS2、ACE2、Alix、TSG101,CD(29,47,63),TLRs(3,6,7),TNF-α,与FCoV感染后的对照来源的EV相比,感染来源的EV和其他人发生了变化。
■我们的研究结果表明,FCoV感染可以改变宿主细胞中EV的产生和组成,影响感染进展和疾病演变。研究与人类密切接触的各种动物冠状病毒中的电动汽车的一个目的是提供有关疾病发展的重要信息,传输,和适应。因此,这项研究表明,电动汽车可以在动物CoV中提供诊断和治疗应用,这种理解可以提供信息,以防止未来的冠状病毒爆发。
UNASSIGNED: Coronavirus (CoV) has become a public health crisis that causes numerous illnesses in humans and certain animals. Studies have identified the small, lipid-bound structures called extracellular vesicles (EVs) as the mechanism through which viruses can enter host cells, spread, and evade the host\'s immune defenses. EVs are able to package and carry numerous viral compounds, including proteins, genetic substances, lipids, and receptor proteins. We proposed that the coronavirus could alter EV production and content, as well as influence EV
biogenesis and composition in host cells.
UNASSIGNED: In the current research, Crandell-Rees feline kidney (CRFK) cells were infected with feline coronavirus (FCoV) in an exosome-free media at a multiplicity of infection (MOI) of 2,500 infectious units (IFU) at 48 h and 72 h time points. Cell viability was analyzed and found to be significantly decreased by 9% (48 h) and 15% (72 h) due to FCoV infection. EVs were isolated by ultracentrifugation, and the surface morphology of isolated EVs was analyzed via Scanning Electron Microscope (SEM).
UNASSIGNED: NanoSight particle tracking analysis (NTA) confirmed that the mean particle sizes of control EVs were 131.9 nm and 126.6 nm, while FCoV infected-derived EVs were 143.4 nm and 120.9 nm at 48 and 72 h, respectively. Total DNA, RNA, and protein levels were determined in isolated EVs at both incubation time points; however, total protein was significantly increased at 48 h. Expression of specific protein markers such as TMPRSS2, ACE2, Alix, TSG101, CDs (29, 47, 63), TLRs (3, 6, 7), TNF-α, and others were altered in infection-derived EVs when compared to control-derived EVs after FCoV infection.
UNASSIGNED: Our findings suggested that FCoV infection could alter the EV production and composition in host cells, which affects the infection progression and disease evolution. One purpose of studying EVs in various animal coronaviruses that are in close contact with humans is to provide significant information about disease development, transmission, and adaptation. Hence, this study suggests that EVs could provide diagnostic and therapeutic applications in animal CoVs, and such understanding could provide information to prevent future coronavirus outbreaks.