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Automated image-based assay for evaluation of HIV neutralization and cell-to-cell fusion inhibition

Sheik-Khalil, Enas LU ; Bray, Mark-Anthony; Özkaya Sahin, Gülsen LU ; Scarlatti, Gabriella; Jansson, Marianne LU ; Carpenter, Anne E. and Fenyö, Eva Maria LU (2014) In BMC Infectious Diseases 14.
Abstract
Background: Standardized techniques to detect HIV-neutralizing antibody responses are of great importance in the search for an HIV vaccine. Methods: Here, we present a high-throughput, high-content automated plaque reduction (APR) assay based on automated microscopy and image analysis that allows evaluation of neutralization and inhibition of cell-cell fusion within the same assay. Neutralization of virus particles is measured as a reduction in the number of fluorescent plaques, and inhibition of cell-cell fusion as a reduction in plaque area. Results: We found neutralization strength to be a significant factor in the ability of virus to form syncytia. Further, we introduce the inhibitory concentration of plaque area reduction (ICpar) as... (More)
Background: Standardized techniques to detect HIV-neutralizing antibody responses are of great importance in the search for an HIV vaccine. Methods: Here, we present a high-throughput, high-content automated plaque reduction (APR) assay based on automated microscopy and image analysis that allows evaluation of neutralization and inhibition of cell-cell fusion within the same assay. Neutralization of virus particles is measured as a reduction in the number of fluorescent plaques, and inhibition of cell-cell fusion as a reduction in plaque area. Results: We found neutralization strength to be a significant factor in the ability of virus to form syncytia. Further, we introduce the inhibitory concentration of plaque area reduction (ICpar) as an additional measure of antiviral activity, i.e. fusion inhibition. Conclusions: We present an automated image based high-throughput, high-content HIV plaque reduction assay. This allows, for the first time, simultaneous evaluation of neutralization and inhibition of cell-cell fusion within the same assay, by quantifying the reduction in number of plaques and mean plaque area, respectively. Inhibition of cell-to-cell fusion requires higher quantities of inhibitory reagent than inhibition of virus neutralization. (Less)
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organization
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type
Contribution to journal
publication status
published
subject
keywords
Automated plaque reduction assay (APR assay), Fluorescence, HIV, Neutralization, Fusion inhibition
in
BMC Infectious Diseases
volume
14
publisher
BioMed Central
external identifiers
  • wos:000343828200001
  • scopus:84922969792
ISSN
1471-2334
DOI
10.1186/1471-2334-14-472
language
English
LU publication?
yes
id
2c385225-31de-428e-8827-e6ba720c97af (old id 4783852)
date added to LUP
2014-12-01 07:37:40
date last changed
2017-01-01 06:18:12
@article{2c385225-31de-428e-8827-e6ba720c97af,
  abstract     = {Background: Standardized techniques to detect HIV-neutralizing antibody responses are of great importance in the search for an HIV vaccine. Methods: Here, we present a high-throughput, high-content automated plaque reduction (APR) assay based on automated microscopy and image analysis that allows evaluation of neutralization and inhibition of cell-cell fusion within the same assay. Neutralization of virus particles is measured as a reduction in the number of fluorescent plaques, and inhibition of cell-cell fusion as a reduction in plaque area. Results: We found neutralization strength to be a significant factor in the ability of virus to form syncytia. Further, we introduce the inhibitory concentration of plaque area reduction (ICpar) as an additional measure of antiviral activity, i.e. fusion inhibition. Conclusions: We present an automated image based high-throughput, high-content HIV plaque reduction assay. This allows, for the first time, simultaneous evaluation of neutralization and inhibition of cell-cell fusion within the same assay, by quantifying the reduction in number of plaques and mean plaque area, respectively. Inhibition of cell-to-cell fusion requires higher quantities of inhibitory reagent than inhibition of virus neutralization.},
  articleno    = {472},
  author       = {Sheik-Khalil, Enas and Bray, Mark-Anthony and Özkaya Sahin, Gülsen and Scarlatti, Gabriella and Jansson, Marianne and Carpenter, Anne E. and Fenyö, Eva Maria},
  issn         = {1471-2334},
  keyword      = {Automated plaque reduction assay (APR assay),Fluorescence,HIV,Neutralization,Fusion inhibition},
  language     = {eng},
  publisher    = {BioMed Central},
  series       = {BMC Infectious Diseases},
  title        = {Automated image-based assay for evaluation of HIV neutralization and cell-to-cell fusion inhibition},
  url          = {http://dx.doi.org/10.1186/1471-2334-14-472},
  volume       = {14},
  year         = {2014},
}