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Type: Journal article
Title: Use of an in vivo FTA assay to assess the magnitude, functional avidity and epitope variant cross-reactivity of T cell responses following HIV-1 recombinant poxvirus vaccination
Author: Wijesundara, D.
Ranasinghe, C.
Jackson, R.
Lidbury, B.
Parish, C.
Quah, B.
Citation: PLoS ONE, 2014; 9(8):e105366-1-e105366-10
Publisher: Public Library of Science (PLoS)
Issue Date: 2014
ISSN: 1932-6203
Statement of
Danushka K. Wijesundara, Charani Ranasinghe, Ronald J. Jackson, Brett A. Lidbury, Christopher R. Parish, Benjamin J. C. Quah
Abstract: Qualitative characteristics of cytotoxic CD8⁺ T cells (CTLs) are important in measuring the effectiveness of CTLs in controlling HIV-1 infections. Indeed, in recent studies patients who are naturally resistant to HIV-1 infections have been shown to possess CTLs that are of high functional avidity and have a high capacity to recognize HIV epitope variants, when compared to HIV-1 infection progressors. When developing efficacious vaccines, assays that can effectively measure CTL quality specifically in vivo are becoming increasingly important. Here we report the use of a recently developed high-throughput multi-parameter technique, known as the fluorescent target array (FTA) assay, to simultaneously measure CTL killing magnitude, functional avidity and epitope variant cross-reactivity in real time in vivo. In the current study we have applied the FTA assay as a screening tool to assess a large cohort of over 20 different HIV-1 poxvirus vaccination strategies in mice. This screen revealed that heterologous poxvirus prime-boost vaccination regimes (i.e., recombinant fowlpox (FPV)-HIV prime followed by a recombinant vaccinia virus (VV)-HIV booster) were the most effective in generating high quality CTL responses in vivo. In conclusion, we have demonstrated how the FTA assay can be utilized as a cost effective screening tool (by reducing the required number of animals by >100 fold), to evaluate a large range of HIV-1 vaccination strategies in terms of CTL avidity and variant cross-reactivity in an in vivo setting.
Keywords: T-Lymphocytes, Cytotoxic
Description: Published August 29, 2014
Rights: © 2014 Wijesundara et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
RMID: 0030087492
DOI: 10.1371/journal.pone.0105366
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Appears in Collections:Medicine publications

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