A novel, sensitive dual-indicator cell line for detection and quantification of inducible, replication-competent latent HIV-1 from reservoir cells
| Autor: | David W. Gludish, Saikat Boliar, Fanny Salasc, Hoi Ping Mok, Mark R. Wills, Isobel Jarvis, David G. Russell, Andrew M. L. Lever |
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| Přispěvatelé: | Salasc, Fanny [0000-0002-6574-1358], Russell, David G [0000-0002-9748-750X], Apollo - University of Cambridge Repository, Russell, David G. [0000-0002-9748-750X], Boliar, Saikat [0000-0002-3340-5368] |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
CD4-Positive T-Lymphocytes viruses 030106 microbiology Human immunodeficiency virus (HIV) lcsh:Medicine HIV Infections 13/109 medicine.disease_cause Virus Replication Virus Cell Line 03 medical and health sciences Gaussia 38/1 Replication (statistics) 631/326/596/1787 medicine Humans Luciferase Latency (engineering) lcsh:Science Luciferases 631/1647/2234 Disease Reservoirs Multidisciplinary Retrovirus biology lcsh:R article virus diseases biology.organism_classification Virology 3. Good health Virus Latency 13/31 030104 developmental biology Viral replication Cell culture 14/63 HIV-1 lcsh:Q Microbiology techniques |
| Zdroj: | Scientific Reports Scientific Reports, Vol 9, Iss 1, Pp 1-10 (2019) |
| Popis: | Understanding the mechanisms involved in HIV infection and latency, and development of a cure, rely on the availability of sensitive research tools such as indicator cells, which allow rigorous quantification of viral activity. Here we describe the construction and validation of a novel dual-indicator cell line, Sup-GGR, which offers two different readouts to quantify viral replication. A construct expressing both Gaussia luciferase and hrGFP in a Tat- and Rev-dependent manner was engineered into SupT1-CCR5 to create Sup-GGR cells. This cell line supports the replication of both X4 and R5-tropic HIV as efficiently as its parental cell line, SupT1-CCR5, and allows repeated sampling without the need to terminate the culture. Sup-GGR demonstrates comparable sensitivity and similar kinetics in virus outgrowth assays (VOA) to SupT1-CCR5 using clinical samples. However the Gaussia luciferase reporter is significantly less labor-intensive and allows earlier detection of reactivated latent viruses compared to the conventional HIV p24 ELISA assay. The Sup-GGR cell line constitutes a versatile new tool for HIV research and clinical trials. |
| Databáze: | OpenAIRE |
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