Longitudinal regulation over time of gene expression in mock infected and HIV-1 infected CD4+ T-cells

HIV-1 infected cells can become a stable reservoir for the virus by harboring latent, but replication competent, viral genomes. Glycolytic metabolism has been identified as a major determinant of susceptibility to HIV-1 infection. We performed microarray analysis to analyze the regulation of transcriptional expression during the transition from productive to latent HIV-1 infection Primary CD4+ T-cells were isolated from total blood of two healthy donors. After isolation CD4+ T-cells were activated for 72 hours by adding the Dynabeads® Human T-Activator CD3/CD28 using a bead/cell ratio of 1:2. Following activation cells were divided in two groups and either infected with HIV-1 NL4-3 (2ng p24/million cells) or mock infected. Cells were cultured in RPMI 1640 supplemented with 20% fetal bovine serum (FBS), penicillin/streptomycin, and 10 ng/mL IL-2 and kept at a concentration between 1-2 million cells/mL. Cell pellets were collected at 7, 9 and 14 days post-infection and used for total RNA extraction and subsequent microarray analysis