目的:细胞外囊泡(EV)是穿梭蛋白质的纳米囊泡,核酸,和脂质,从而影响细胞行为。最近的一系列报告表明,电动汽车参与了感染性生物学,影响宿主免疫并在病毒生命周期中发挥作用。在目前的工作中,我们调查了EV介导的乙型肝炎病毒(HBV)感染的传播。
方法:我们通过使用HBV感染培养系统,使用源自人源化嵌合小鼠(PXB细胞)的原代人肝细胞,研究了EV介导的HBV感染传播。通过超速离心分离纯化的EV。为了分析电动汽车和病毒体,我们使用受激发射耗尽显微镜。
结果:来自HBV感染的PXB细胞的纯化EV被证明含有HBVDNA,并且能够将HBVDNA传递给幼稚的PXB细胞。这些HBV-DNA传输EV被证明是通过神经酰胺触发的EV产生途径产生的。此外,我们表明,这些HBV-DNA传输EV抗抗体中和;受激发射耗尽显微镜显示,电动汽车缺乏乙型肝炎表面抗原,中和抗体的靶标。
结论:这些发现表明,电动汽车藏有能够在HBV感染期间将病毒DNA传递到肝细胞的DNA货物,代表HBV感染的另一种抗体中和耐药途径。
OBJECTIVE: An extracellular vesicle (EV) is a nanovesicle that shuttles proteins, nucleic acids, and lipids, thereby influencing cell behavior. A recent crop of reports have shown that EVs are involved in infectious biology, influencing host immunity and playing a role in the viral life cycle. In the present work, we investigated the EV-mediated transmission of hepatitis B virus (HBV) infection.
METHODS: We investigated the EV-mediated transmission of HBV infection by using a HBV infectious culture system that uses primary human hepatocytes derived from humanized chimeric mice (PXB-cells). Purified EVs were isolated by ultracentrifugation. To analyze the EVs and virions, we used stimulated emission depletion microscopy.
RESULTS: Purified EVs from HBV-infected PXB-cells were shown to contain HBV DNA and to be capable of transmitting HBV DNA to naive PXB-cells. These HBV-DNA-transmitting EVs were shown to be generated through a ceramide-triggered EV production pathway. Furthermore, we showed that these HBV-DNA-transmitting EVs were resistant to antibody neutralization; stimulated emission depletion microscopy showed that EVs lacked hepatitis B surface antigen, the target of neutralizing antibodies.
CONCLUSIONS: These findings suggest that EVs harbor a DNA cargo capable of transmitting viral DNA into hepatocytes during HBV infection, representing an additional antibody-neutralization-resistant route of HBV infection.