Automated image-based assay for evaluation of HIV neutralization and cell-to-cell fusion inhibition
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/4783852
- author
- Sheik-Khalil, Enas LU ; Bray, Mark-Anthony ; Özkaya Sahin, Gülsen LU ; Scarlatti, Gabriella ; Jansson, Marianne LU ; Carpenter, Anne E. and Fenyö, Eva Maria LU
- organization
- publishing date
- 2014
- 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
- article number
- 472
- publisher
- BioMed Central (BMC)
- external identifiers
-
- wos:000343828200001
- scopus:84922969792
- pmid:25176034
- 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
- 2016-04-01 14:33:30
- date last changed
- 2022-01-28 01:13:59
@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.}},
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}},
keywords = {{Automated plaque reduction assay (APR assay); Fluorescence; HIV; Neutralization; Fusion inhibition}},
language = {{eng}},
publisher = {{BioMed Central (BMC)}},
series = {{BMC Infectious Diseases}},
title = {{Automated image-based assay for evaluation of HIV neutralization and cell-to-cell fusion inhibition}},
url = {{https://lup.lub.lu.se/search/files/4037005/5464982.pdf}},
doi = {{10.1186/1471-2334-14-472}},
volume = {{14}},
year = {{2014}},
}