A scalable workflow to test “shock and kill” therapeutic approaches against the HIV-1 latent reservoir in blood cells ex vivo
Summary: 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”) a...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723002113 |
| _version_ | 1797839084555599872 |
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| author | Francesco Andrea Procopio Andrea Savarino Iart Luca Shytaj |
| author_facet | Francesco Andrea Procopio Andrea Savarino Iart Luca Shytaj |
| author_sort | Francesco Andrea Procopio |
| collection | DOAJ |
| description | Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-09T15:52:31Z |
| format | Article |
| id | doaj.art-f8be3c6834fe4ae09552feab9061ada7 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-09T15:52:31Z |
| publishDate | 2023-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-f8be3c6834fe4ae09552feab9061ada72023-04-26T06:00:49ZengElsevierSTAR Protocols2666-16672023-06-0142102253A scalable workflow to test “shock and kill” therapeutic approaches against the HIV-1 latent reservoir in blood cells ex vivoFrancesco Andrea Procopio0Andrea Savarino1Iart Luca Shytaj2Services of Immunology and Allergy, Lausanne University Hospital, University of Lausanne, 1011 Lausanne, Switzerland; Corresponding authorDepartment of Infectious Diseases, Italian Institute of Health, 00161 Rome, ItalyFaculty of Life Sciences, School of Cellular and Molecular Medicine, University of Bristol, BS8 1TD Bristol, UK; Corresponding authorSummary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723002113Cell BiologyCell IsolationCell-based AssaysCell Separation/FractionationImmunologyMicrobiology |
| spellingShingle | Francesco Andrea Procopio Andrea Savarino Iart Luca Shytaj A scalable workflow to test “shock and kill” therapeutic approaches against the HIV-1 latent reservoir in blood cells ex vivo STAR Protocols Cell Biology Cell Isolation Cell-based Assays Cell Separation/Fractionation Immunology Microbiology |
| 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 | Cell Biology Cell Isolation Cell-based Assays Cell Separation/Fractionation Immunology Microbiology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723002113 |
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