mRNA expression profiles in productively HIV-1 infected and bystander primary human CD4+ T-cells
CD4+ T-cells are the main target of HIV-1 and several host factors can positively or negatively modulate HIV-1 infection of these cells. MiRNAs aresmall regulatory RNAs that are involved in the regulation of basic cellular functions. They are also increasingly recognized as host factors regulatingHIV-1 infection, replication and persistence. In order to identify miRNAs involved in HIV-1 infection of CD4+ T-cells, we performed globaltranscriptomic analyses of productively infected and HIV-1 exposed but non infected bystander CD4+ T-cells and compared their MIRNA profiles with uninfected cells. Theseanalyses were performed in CD4+T-cells isolated from 3 different healthy blood donors. Both miRNA and mRNA expression profiles were compared. Our results show that bystander and uninfected CD4+ T-cells do not display important differences in their miRNA expression profiles even though their respectivemRNA expression profiles were markedly different. In contrast, both mRNA and miRNA expressionprofiles from productively infected CD4+ T-cells were significantly different from those of uninfected cells. Overall, these results suggest that HIV-1infection impacts the miRNA expression profile of primary CD4+ T-cells. Overall design: Human CD4+ T-cells from 3 different healthy blood donors were isolated from peripheral blood mononuclear cells (PBMCs) and activated for 3 days in the presence of anti-CD3/anti-CD28 and IL-2 in RPMI+10% fetal bovine serum (FBS). Activated T-cells were infected with GFP-marked HIV-1 (strain NL4.3-ADA-IRES-GFP) for 48h at a multiplicity of infection (MOI)=2. Productively infected (GFP+) and bystander cells (GFP-) were sorted by FACS and their total RNA extracted using the RNeasy kit from QIAGEN.