Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/82353
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

SCOPUSTM   
Citations 20

10
Updated on Jun 7, 2024

Web of ScienceTM
Citations 20

8
Updated on Oct 27, 2023

Page view(s) 50

543
Updated on Jun 13, 2024

Download(s) 50

146
Updated on Jun 13, 2024

Google ScholarTM

Check

Altmetric


Plumx

Items in DR-NTU are protected by copyright, with all rights reserved, unless otherwise indicated.