NIH Research Festival
Natural antisense transcripts (NATs), which are transcribed from the opposite strand of protein-coding genes, regulate gene expression through epigenetic, post-transcriptional, and/or post-translational modifications. The human immunodeficiency virus type I (HIV-1) has been shown to express one or more NATs from a promoter within the 3‚Äô long terminal repeat. HIV-1 antisense transcripts (Ast) are capable of inducing proviral latency in in vitro cell culture. Our studies will identify and quantify HIV-1 Ast in vivo. We developed and optimized a digital PCR-based assay to measure levels of Ast in small pools of infected cells using the ACH2 cell line. ACH2 cells each carry a single copy of an integrated HIV- 1 provirus that expresses sense and antisense RNA at low levels. In 6 experimental replicates of 50 ACH2 cells, our digital PCR approach detected HIV-1 Ast in an average of 36% of the cells (range 22%-70%), consistent with previous studies. We next applied our new assay to PBMC collected from a donor with HIV- 1 and on antiretroviral therapy. Testing 2 pools of aliquots of about 22 infected PBMC from this donor, we found that up to 54% of the infected cells contain HIV-1 Ast. These data demonstrate that Ast is expressed in people living with HIV-1 and lead to the question of whether HIV-1 Ast expression can contribute to viral latency and persistence during treatment. In our future studies, we will investigate varying levels of plasma viremia in a cohort of 50 people living with HIV-1.
Scientific Focus Area: Virology
This page was last updated on Monday, September 25, 2023