Quantum dots(QDs)-based single particle analysis technique enables real-time tracking of the viral infection in live cells with great sensitivity over a long period of time.The porcine reproductive and respiratory syn...Quantum dots(QDs)-based single particle analysis technique enables real-time tracking of the viral infection in live cells with great sensitivity over a long period of time.The porcine reproductive and respiratory syndrome virus(PRRSV)is a small virus with the virion size of 40–60 nm which causes great economic losses to the swine industry worldwide.A clear understanding of the viral infection mechanism is essential for the development of effective antiviral strategies.In this study,we labeled the PRRSV with QDs using the streptavidin–biotin labeling system and monitored the viral infection process in live cells.Our results indicated that the labeling method had negligible effect on viral infectivity.We also observed that prior to the entry,PRRSV vibrated on the plasma membrane,and entered the cells via endosome mediated cell entry pathway.Viruses moved in a slow–fast–slow oscillatory movement pattern and finally accumulated in a perinuclear region of the cell.Our results also showed that once inside the cell,PRRSV moved along the microtubule,microfilament and vimentin cytoskeletal elements.During the transport process,virus particles also made contacts with non-muscle myosin heavy chainⅡ-A(NMHCⅡ-A),visualized as small spheres in cytoplasm.This study can facilitate the application of QDs in virus infection imaging,especially the smaller-sized viruses and provide some novel and important insights into PRRSV infection mechanism.展开更多
基金support from the National Natural Science Foundation of China(Grant Nos.31570151 and 31490601)the Program for Science and Technology Innovation Talents in Universities of Henan Province(Grant No.17HASTIT039)+1 种基金the Key Scientific Research Project of Henan Province Higher Education(16A180044)the Open Research Fund Program of the State Key Laboratory of Virology of China(Grant No.2017KF005)。
文摘Quantum dots(QDs)-based single particle analysis technique enables real-time tracking of the viral infection in live cells with great sensitivity over a long period of time.The porcine reproductive and respiratory syndrome virus(PRRSV)is a small virus with the virion size of 40–60 nm which causes great economic losses to the swine industry worldwide.A clear understanding of the viral infection mechanism is essential for the development of effective antiviral strategies.In this study,we labeled the PRRSV with QDs using the streptavidin–biotin labeling system and monitored the viral infection process in live cells.Our results indicated that the labeling method had negligible effect on viral infectivity.We also observed that prior to the entry,PRRSV vibrated on the plasma membrane,and entered the cells via endosome mediated cell entry pathway.Viruses moved in a slow–fast–slow oscillatory movement pattern and finally accumulated in a perinuclear region of the cell.Our results also showed that once inside the cell,PRRSV moved along the microtubule,microfilament and vimentin cytoskeletal elements.During the transport process,virus particles also made contacts with non-muscle myosin heavy chainⅡ-A(NMHCⅡ-A),visualized as small spheres in cytoplasm.This study can facilitate the application of QDs in virus infection imaging,especially the smaller-sized viruses and provide some novel and important insights into PRRSV infection mechanism.
