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Title: | Evidence for a biphasic mode of respiratory syncytial virus transmission in permissive HEp2 cell monolayers | Authors: | Huong, Tra Nguyen Ravi, Laxmi Iyer Tan, Boon Huan Sugrue, Richard J. |
Keywords: | Biological Sciences | Issue Date: | 2016 | Source: | Huong, T. N., Ravi, L. I., Tan, B. H., & Sugrue, R. J. (2016). Evidence for a biphasic mode of respiratory syncytial virus transmission in permissive HEp2 cell monolayers. Virology Journal, 13, 12-. | Series/Report no.: | Virology Journal | Abstract: | Background: During respiratory syncytial virus (RSV) infection filamentous virus particles are formed on the cell surface. Although the virus infectivity remains cell-associated, low levels of cell-free virus is detected during advanced infection. It is currently unclear if this cell-free virus infectivity is due to a low-efficiency specific cell-release mechanism, or if it arises due to mechanical breakage following virus-induced cell damage at the advanced stage of infection. Understanding the origin of this cell-free virus is a prerequisite for understanding the mechanism of RSV transmission in permissive cells. In this study we describe a detailed examination of RSV transmission in permissive HEp2 cell monolayers. Methods: HEp2 cell monolayers were infected with RSV using a multiplicity of infection of 0.0002, and the course of infection monitored over 5 days. The progression of the virus infection within the cell monolayers was performed using bright-field microscopy to visualise the cell monolayer and immunofluorescence microscopy to detect virus-infected cells. The cell-associated and cell-free virus infectivity were determined by virus plaque assay, and the virus-induced cell cytotoxicity determined by measuring cell membrane permeability and cellular DNA fragmentation. Results: At 2 days-post infection (dpi), large clusters of virus-infected cells could be detected indicating localised transmission in the cell monolayer, and during this stage we failed to detect either cell-free virus or cell cytotoxicity. At 3 dpi the presence of much larger infected cell clusters correlated with the begining of virus-induced changes in cell permeability. The presence of cell-free virus correlated with continued increase in cell permeability and cytotoxicity at 4 and 5 dpi. At 5 dpi extensive cell damage, syncytial formation, and increased cellular DNA fragmentation was noted. However, even at 5 dpi the cell-free virus constituted less than 1 % of the total virus infectivity. Conclusions: Our data supports a model of RSV transmission that initially involves the localised cell-to-cell spread of virus particles within the HEp2 cell monolayer. However, low levels of cell free-virus infectivity was observed at the advanced stages of infection, which correlated with a general loss in cell monolayer integrity due to virus-induced cytotoxicity. | URI: | https://hdl.handle.net/10356/82353 http://hdl.handle.net/10220/39945 |
ISSN: | 1743-422X | DOI: | 10.1186/s12985-016-0467-9 | Schools: | School of Biological Sciences | Rights: | © 2016 Huong et al. Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. | Fulltext Permission: | open | Fulltext Availability: | With Fulltext |
Appears in Collections: | SBS Journal Articles |
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Evidence for a biphasic mode of respiratory syncytial virus transmission in permissive HEp2 cell monolayers Negative-strand RNA viruses.pdf | 2.08 MB | Adobe PDF | View/Open |
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