| Autor: |
Salasc F; Department of Medicine, University of Cambridge, Cambridge, UK., Gludish DW; Cornell University College of Veterinary Medicine, New York, USA., Jarvis I; Department of Medicine, University of Cambridge, Cambridge, UK., Boliar S; Cornell University College of Veterinary Medicine, New York, USA., Wills MR; Department of Medicine, University of Cambridge, Cambridge, UK., Russell DG; Cornell University College of Veterinary Medicine, New York, USA. dgr8@cornell.edu., Lever AML; Department of Medicine, University of Cambridge, Cambridge, UK. amll1@medschl.cam.ac.uk.; Yong Loo Lin School of Medicine National University of Singapore, Singapore, Singapore. amll1@medschl.cam.ac.uk., Mok HP; Department of Medicine, University of Cambridge, Cambridge, UK. hpm22@cam.ac.uk. |
| Abstrakt: |
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. |
