Drug‐associated DNA Damage in HIV‐1 Patients taking HAART

Abstract

Despite the reduction in mortality in HIV‐1 positive patients due to the use of highly active antiretroviral therapy (HAART), adverse effects have been associated to long‐term usage. Recently, several studies have demonstrated that HIV‐1 infected patients are under chronic oxidative stress caused by an inflammatory response to the latent infection, or as an adverse effect of the antiretroviral therapy. The aim of this study was to evaluate the DNA damage in peripheral blood mononuclear cells of HIV‐1 patients treated with HAART when compared to healthy donors. Frozen peripheral blood mononuclear cells (PBMCs) obtained from HIV‐1 positive subjects with undetectable viremia and under HAART were used for this study. An aliquot of the thawed cells was used for DNA isolation and the remaining cells were cultured for seven days. Single and double‐strand DNA breaks were evaluated using an alkaline gel electrophoresis (single‐strand gel electrophoresis) and the neutral comet assay, respectively. PBMCs from healthy donors were used as negative controls. EMS (ethylmethane sulfonate) was used as a positive control. Interestingly, the alkaline gel showed higher DNA fragmentation in HIV‐1 positive samples when compared to an HIV‐negative control and to a sample treated with EMS. Additionally, the neutral comet assay showed presence of double‐stranded DNA breaks in HIV‐1 positive cells when compared to a healthy donor sample. The extent of the single or double‐stranded DNA breaks varied among samples depending on type of therapy and the length of time under the therapy. Taken together, our results suggest that HAART may affect the oxidative stress imbalance in HIV‐1 positive patients.