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Imaging Early Steps of Sindbis Virus Infection by Total Internal Reflection Fluorescence Microscopy
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 sin...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi Publishing Corporation
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3265306/ https://www.ncbi.nlm.nih.gov/pubmed/22312345 http://dx.doi.org/10.1155/2011/535206 |
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author | Gu, Youling Yang, Yuanzheng Liu, Yuechueng |
author_facet | Gu, Youling Yang, Yuanzheng Liu, Yuechueng |
author_sort | Gu, Youling |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3265306 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Hindawi Publishing Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-32653062012-02-06 Imaging Early Steps of Sindbis Virus Infection by Total Internal Reflection Fluorescence Microscopy Gu, Youling Yang, Yuanzheng Liu, Yuechueng Adv Virol Research Article 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. Hindawi Publishing Corporation 2011 2011-11-24 /pmc/articles/PMC3265306/ /pubmed/22312345 http://dx.doi.org/10.1155/2011/535206 Text en Copyright © 2011 Youling Gu et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Gu, Youling Yang, Yuanzheng Liu, Yuechueng Imaging Early Steps of Sindbis Virus Infection by Total Internal Reflection Fluorescence Microscopy |
title | Imaging Early Steps of Sindbis Virus Infection by Total Internal Reflection Fluorescence Microscopy |
title_full | Imaging Early Steps of Sindbis Virus Infection by Total Internal Reflection Fluorescence Microscopy |
title_fullStr | Imaging Early Steps of Sindbis Virus Infection by Total Internal Reflection Fluorescence Microscopy |
title_full_unstemmed | Imaging Early Steps of Sindbis Virus Infection by Total Internal Reflection Fluorescence Microscopy |
title_short | Imaging Early Steps of Sindbis Virus Infection by Total Internal Reflection Fluorescence Microscopy |
title_sort | imaging early steps of sindbis virus infection by total internal reflection fluorescence microscopy |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3265306/ https://www.ncbi.nlm.nih.gov/pubmed/22312345 http://dx.doi.org/10.1155/2011/535206 |
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