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Modification and optimization of the inhibition of HIV-1 cell-to-cell transmission assay

HIV-1 infection is caused by cell-free and cell-associated viruses. Currently most of the assays used to screen potential HIV-1 entry inhibitors focus on the inhibition of cell-free viruses. One assay that is widely employed is the TZM-bl neutralization assay that uses pseudotyped viruses. However,...

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Autores principales: Alexandre, Kabamba, Malatji, Kanyane, Mulaudzi, Takalani
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7475115/
https://www.ncbi.nlm.nih.gov/pubmed/32923375
http://dx.doi.org/10.1016/j.mex.2020.101014
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author Alexandre, Kabamba
Malatji, Kanyane
Mulaudzi, Takalani
author_facet Alexandre, Kabamba
Malatji, Kanyane
Mulaudzi, Takalani
author_sort Alexandre, Kabamba
collection PubMed
description HIV-1 infection is caused by cell-free and cell-associated viruses. Currently most of the assays used to screen potential HIV-1 entry inhibitors focus on the inhibition of cell-free viruses. One assay that is widely employed is the TZM-bl neutralization assay that uses pseudotyped viruses. However, a study by Abela et al. showed that many inhibitors that potently inhibit cell-free HIV-1 in this assay can be less effective against the cell-to-cell transmission of the virus. These researchers then designed a method to screen entry inhibitors for activity against cell-associated HIV-1, using pseudotyped viruses. The main limitation of this method, however, was that it can only be reliably employed against viruses that cannot infect target cells as cell-free virion in the absence of a polycation supplement such as DEAE (diethylaminoethyl). Thus, in the current study we provide modifications to this method that solves the problem and makes it possible to study entry inhibitors against cell-to-cell infection of both polycation depend and independent viruses. The main modification involves the introduction of the relative light unit (RLU) vs. virus producing 293-T cells / corresponding supernatants graph. This graph is used to select a virus input that only allows for the detection of cell-associated viruses infection. • The method is a modification of the cell-to-cell transmission assay published by Abela et al. • The method allows for the study of the inhibition of cell-to-cell transmission of both polycation dependent and independent HIV-1 pseudoviruses.
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spelling pubmed-74751152020-09-11 Modification and optimization of the inhibition of HIV-1 cell-to-cell transmission assay Alexandre, Kabamba Malatji, Kanyane Mulaudzi, Takalani MethodsX Agricultural and Biological Science HIV-1 infection is caused by cell-free and cell-associated viruses. Currently most of the assays used to screen potential HIV-1 entry inhibitors focus on the inhibition of cell-free viruses. One assay that is widely employed is the TZM-bl neutralization assay that uses pseudotyped viruses. However, a study by Abela et al. showed that many inhibitors that potently inhibit cell-free HIV-1 in this assay can be less effective against the cell-to-cell transmission of the virus. These researchers then designed a method to screen entry inhibitors for activity against cell-associated HIV-1, using pseudotyped viruses. The main limitation of this method, however, was that it can only be reliably employed against viruses that cannot infect target cells as cell-free virion in the absence of a polycation supplement such as DEAE (diethylaminoethyl). Thus, in the current study we provide modifications to this method that solves the problem and makes it possible to study entry inhibitors against cell-to-cell infection of both polycation depend and independent viruses. The main modification involves the introduction of the relative light unit (RLU) vs. virus producing 293-T cells / corresponding supernatants graph. This graph is used to select a virus input that only allows for the detection of cell-associated viruses infection. • The method is a modification of the cell-to-cell transmission assay published by Abela et al. • The method allows for the study of the inhibition of cell-to-cell transmission of both polycation dependent and independent HIV-1 pseudoviruses. Elsevier 2020-07-26 /pmc/articles/PMC7475115/ /pubmed/32923375 http://dx.doi.org/10.1016/j.mex.2020.101014 Text en © 2020 Published by Elsevier B.V. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Agricultural and Biological Science
Alexandre, Kabamba
Malatji, Kanyane
Mulaudzi, Takalani
Modification and optimization of the inhibition of HIV-1 cell-to-cell transmission assay
title Modification and optimization of the inhibition of HIV-1 cell-to-cell transmission assay
title_full Modification and optimization of the inhibition of HIV-1 cell-to-cell transmission assay
title_fullStr Modification and optimization of the inhibition of HIV-1 cell-to-cell transmission assay
title_full_unstemmed Modification and optimization of the inhibition of HIV-1 cell-to-cell transmission assay
title_short Modification and optimization of the inhibition of HIV-1 cell-to-cell transmission assay
title_sort modification and optimization of the inhibition of hiv-1 cell-to-cell transmission assay
topic Agricultural and Biological Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7475115/
https://www.ncbi.nlm.nih.gov/pubmed/32923375
http://dx.doi.org/10.1016/j.mex.2020.101014
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