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Imaging Early Steps of Sindbis Virus Infection by Total Internal Reflection Fluorescence Microscopy

机译:全内反射荧光显微镜成像的信德比斯病毒感染的早期步骤。

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摘要

Sindbis virus (SINV) is an alphavirus that has a broad host range and has been widely used as a vector for recombinant gene transduction, DNA-based vaccine production, and oncolytic cancer therapy. The mechanism of SINV entry into host cells has yet to be fully understood. In this paper, we used single virus tracking under total internal reflection fluorescence microscopy (TIRFM) to investigate SINV attachment to cell surface. Biotinylated viral particles were labeled with quantum dots, which retained viral viability and infectivity. By time-lapse imaging, we showed that the SINV exhibited a heterogeneous dynamics on the surface of the host cells. Analysis of SINV motility demonstrated a two-step attachment reaction. Moreover, dual color TIRFM of GFP-Rab5 and SINV suggested that the virus was targeted to the early endosomes after endocytosis. These findings demonstrate the utility of quantum dot labeling in studying the early steps and behavior of SINV infection.
机译:Sindbis病毒(SINV)是一种具有广泛宿主范围的alpha病毒,已被广泛用作重组基因转导,基于DNA的疫苗生产和溶瘤性癌症治疗的载体。 SINV进入宿主细胞的机制尚未完全了解。在本文中,我们使用全内反射荧光显微镜(TIRFM)下的单病毒跟踪研究SINV与细胞表面的附着。用量子点标记生物素化的病毒颗粒,该量子点保留了病毒的生存力和感染力。通过延时成像,我们表明SINV在宿主细胞的表面上表现出异质的动力学。对SINV运动性的分析表明了两步附着反应。此外,GFP-Rab5和SINV的双色TIRFM表明该病毒靶向内吞后的早期内体。这些发现证明了量子点标记在研究SINV感染的早期步骤和行为方面的实用性。

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