Figure 3
Comparative analysis of Tat-mediated gene modulation in iDC and MDM. A. Microarray RNA levels in MDM infected with Ad-TatSF2 or Ad-TatHXB2 (left panel) and in MDM infected with Ad-TatHXB2 for 30 h or with HIVBal for 7 and 10 days (right panel). B. Venn diagrams representing total number of genes modulated by Tat or HIV-1 in MDM and the overlapping subset similarly modulated in the two conditions. C. Modulation of a subset of ISG in MDM evaluated by quantitative RT-PCR. TatSF2 and TatSF272 were expressed using an adenoviral vector and RNA was purified 16 and 30 h after infection. TatBal, and TatBal1-72 were expressed using a lentiviral vector and RNA was purified 30 and 48 h after infection from cells sorted 24 h after infection using a GFP maker present in the lentivector. Results are reported as the average (n-fold) induction of three independent experiments relative to that of the control sample infected with the background vector. Values are normalized using 18S RNA and Tat levels. Significant differences between the cumulative levels of all the evaluated ISG expressed in TatSF2 and in TatSF21-72 were detected at 16 and 30 h (p < 0.0001 for both time points, T-test) and also between TatBal, and TatBal1-72 at 30 (p = 0.0004) and 48 h (p < 0.0001). Significant differences could also be detected when individual gene expression was compared. P values for gene fold inductions in TatSF21-72 samples compared to TatSF2 were: IRF7-16 h: p = 0.03, IRF7-30 h: p = 0.006; STAT1-16 h: p = 0.04, STAT1-30 h: p = 0.03; IP10-30 h: p = 0.04; MIG-16 h: p = 0.0001, MIG-30 h: p = 0.03; TRAIL-16 h: p = 0.04; ISG20-16 h: p = 0.02, ISG20-30 h: p = 0.002; CCL20-16 h: p = 0.003, CCL20-30 h: p = 0.03; IFI35-16 h and 30 h: p = 0.03; RSAD2-30 h: p = 0.01; OASL-16 h: p = 0.01, OASL-30 h: p = 0.008. P values for gene fold inductions in the TatBal samples compared to the TatBal1-72 were: MCP3-30 h: p = 0.008; TRAIL-30 h: p = 0.002; ISG20-30 h: p = 0.01; IFI35-30 h: p = 0.05, IRF7-48 h: p = 0.02; STAT1-48 h: p = 0.02; MIG-48 h: p = 0.02. D. Number of genes modulated by Tat and HIV-1 in MDM and in iDC and subset similarly modulated in the two cell types.