Cargando…
A scalable workflow to test “shock and kill” therapeutic approaches against the HIV-1 latent reservoir in blood cells ex vivo
Integrated HIV-1 DNA persists in cells of people living with HIV during antiretroviral treatment, but its quantification is hindered by its rarity. Here, we present an optimized protocol to evaluate “shock and kill” therapeutic strategies, including both the latency reactivation (“shock”) and elimin...
Autores principales: | , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10182335/ https://www.ncbi.nlm.nih.gov/pubmed/37097819 http://dx.doi.org/10.1016/j.xpro.2023.102253 |
_version_ | 1785041749380431872 |
---|---|
author | Procopio, Francesco Andrea Savarino, Andrea Shytaj, Iart Luca |
author_facet | Procopio, Francesco Andrea Savarino, Andrea Shytaj, Iart Luca |
author_sort | Procopio, Francesco Andrea |
collection | PubMed |
description | Integrated HIV-1 DNA persists in cells of people living with HIV during antiretroviral treatment, but its quantification is hindered by its rarity. Here, we present an optimized protocol to evaluate “shock and kill” therapeutic strategies, including both the latency reactivation (“shock”) and elimination of infected cells (“kill”) stages. We describe steps for the sequential use of nested PCR-based assays and viability sorting to allow for scalable and rapid screening of candidate therapeutics in patient-derived blood cells. For complete details on the use and execution of this protocol, please refer to Shytaj et al..(1) |
format | Online Article Text |
id | pubmed-10182335 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-101823352023-05-14 A scalable workflow to test “shock and kill” therapeutic approaches against the HIV-1 latent reservoir in blood cells ex vivo Procopio, Francesco Andrea Savarino, Andrea Shytaj, Iart Luca STAR Protoc Protocol Integrated HIV-1 DNA persists in cells of people living with HIV during antiretroviral treatment, but its quantification is hindered by its rarity. Here, we present an optimized protocol to evaluate “shock and kill” therapeutic strategies, including both the latency reactivation (“shock”) and elimination of infected cells (“kill”) stages. We describe steps for the sequential use of nested PCR-based assays and viability sorting to allow for scalable and rapid screening of candidate therapeutics in patient-derived blood cells. For complete details on the use and execution of this protocol, please refer to Shytaj et al..(1) Elsevier 2023-04-24 /pmc/articles/PMC10182335/ /pubmed/37097819 http://dx.doi.org/10.1016/j.xpro.2023.102253 Text en © 2023 The Authors https://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 | Protocol Procopio, Francesco Andrea Savarino, Andrea Shytaj, Iart Luca A scalable workflow to test “shock and kill” therapeutic approaches against the HIV-1 latent reservoir in blood cells ex vivo |
title | A scalable workflow to test “shock and kill” therapeutic approaches against the HIV-1 latent reservoir in blood cells ex vivo |
title_full | A scalable workflow to test “shock and kill” therapeutic approaches against the HIV-1 latent reservoir in blood cells ex vivo |
title_fullStr | A scalable workflow to test “shock and kill” therapeutic approaches against the HIV-1 latent reservoir in blood cells ex vivo |
title_full_unstemmed | A scalable workflow to test “shock and kill” therapeutic approaches against the HIV-1 latent reservoir in blood cells ex vivo |
title_short | A scalable workflow to test “shock and kill” therapeutic approaches against the HIV-1 latent reservoir in blood cells ex vivo |
title_sort | scalable workflow to test “shock and kill” therapeutic approaches against the hiv-1 latent reservoir in blood cells ex vivo |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10182335/ https://www.ncbi.nlm.nih.gov/pubmed/37097819 http://dx.doi.org/10.1016/j.xpro.2023.102253 |
work_keys_str_mv | AT procopiofrancescoandrea ascalableworkflowtotestshockandkilltherapeuticapproachesagainstthehiv1latentreservoirinbloodcellsexvivo AT savarinoandrea ascalableworkflowtotestshockandkilltherapeuticapproachesagainstthehiv1latentreservoirinbloodcellsexvivo AT shytajiartluca ascalableworkflowtotestshockandkilltherapeuticapproachesagainstthehiv1latentreservoirinbloodcellsexvivo AT procopiofrancescoandrea scalableworkflowtotestshockandkilltherapeuticapproachesagainstthehiv1latentreservoirinbloodcellsexvivo AT savarinoandrea scalableworkflowtotestshockandkilltherapeuticapproachesagainstthehiv1latentreservoirinbloodcellsexvivo AT shytajiartluca scalableworkflowtotestshockandkilltherapeuticapproachesagainstthehiv1latentreservoirinbloodcellsexvivo |