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Updated: Aug 8, 2026
Humanized NOD/SCID/IL2rγnull (hu-NSG) Mouse Model for HIV Replication and Latency Studies
Published on: January 7, 2019
Uptake of 2',3'-dideoxyadenosine in human immunodeficiency virus-infected and noninfected human cells
R P Agarwal1, M E Busso, A M Mian
1Department of Oncology, University of Miami School of Medicine, FL 33101.
Insights
The uptake of 2
Area of Science:
- Pharmacology
- Cell Biology
- Virology
Background:
- 2',3'-dideoxyadenosine is an antiviral nucleoside analog.
- Understanding its cellular uptake is crucial for developing effective HIV therapies.
Purpose of the Study:
- To investigate the cellular uptake mechanisms of 2',3'-dideoxyadenosine in various human cell types.
- To compare uptake patterns in HIV-infected and uninfected cells, and in cell lines versus primary cells.
Main Methods:
- Incubation of H9 cells (T cell line), Namalwa cells (B cell line), and peripheral blood mononuclear cells (PBMCs) with radiolabeled 2',3'-dideoxyadenosine.
- Assessment of intracellular vs. extracellular radioactivity at different temperatures.
- Inhibition studies using various nucleosides and enzyme inhibitors.
- Analysis of dideoxyadenosine triphosphate formation.
Main Results:
- Cellular uptake of 2',3'-dideoxyadenosine was rapid and temperature-dependent, suggesting diffusion.
- Inhibition patterns varied between H9 cells and PBMCs.
- Dideoxyadenosine triphosphate accumulation did not consistently correlate with uptake in H9 cells but did in PBMCs.
Conclusions:
- The primary mechanism for 2',3'-dideoxyadenosine uptake appears to be simple diffusion.
- Significant metabolic differences exist between cell lines and primary PBMCs.
- These cellular distinctions are important for optimizing anti-HIV drug development.
Abstract:
The uptake of 2',3'-dideoxyadenosine was examined in a human immunodeficiency virus (HIV) infected and uninfected T cell line (H9 cells), a B cell line (Namalwa), and in normal peripheral blood mononuclear cells. After a 10-minute incubation at ambient temperature, the intracellular 2',3'-dideoxyadenosine-derived radioactivity was 8- to 16-fold higher than the extracellular radioactivity. In metabolically inactive cells (0 degrees C), the intracellular and extracellular 2',3'-dideoxyadenosine-derived radioactivities were nearly equal. In infected and noninfected H9 cells, a large excess of p-nitrobenzylmercaptopurine riboside or pyrimidine nucleosides weakly inhibited the uptake of 2',3'-dideoxyadenosine (7-30%), whereas deoxycoformycin was a stronger inhibitor (50-80%). Purine nucleosides minimally enhanced the uptake (10-20%). The cellular uptake was not associated with the accumulation of dideoxyadenosine triphosphate. In normal peripheral blood mononuclear cells, the uptake of 2',3'-dideoxyadenosine was inhibited by all agents except 2'-deoxyadenosine (15% enhancement). In contrast to H9 cells, the formation and accumulation of dideoxyadenosine triphosphate paralleled the uptake of dideoxyadenosine. The results of these studies suggest that the major route of transport of 2',3'-dideoxyadenosine into cells is by simple diffusion and that different metabolic patterns exist among cell lines and normal peripheral blood mononuclear cells. An understanding of these cellular differences could aid in the development of therapeutic strategies directed against HIV.
