Figure 6

The effects of transduction with lentiviral vector HR-Hutat2 on the gene expression of human macrophage-related functional and regulatory genes and on kinetics of pro-inflammatory cytokines IL1β, IL8, IL10, and TNF-α. Human monocyte-derived macrophages (hMDM) were differentiated from isolated peripheral blood mononuclear cells in M-CSF-containing medium. On day 7 and day 8 in vitro (DIV 7 and DIV 8), hMDMs were transduced with HR-Hutat2 vector at a MOI of 10 or 50. Total RNA was extracted from non-transduced hMDM (Normal) and transduced hMDM on day 9 post-transduction. Cell culture mediums were collected every 3 days post-transduction. (A) Comparative analysis of the transcriptional profiling of 15 hMDM-related functional and regulatory genes by qRT-PCR. Among the 15 genes, only the transcription of IL8, STAT1, and IDO1 genes changed. (B-E) Sequential changes of IL1β, IL10, IL8, and TNF-α levels in the supernatants of normal and transduced hMDMs at a MOI of 10 or 50. Normal, Non-transduced hMDM; MOI 10, hMDM transduced with HR-Hutat2 at the MOI of 10; MOI 50, hMDM transduced with HR-Hutat2 at the MOI of 50. *P <0.01, ^# P <0.05 compared with normal. Results shown represent mean values from three independent experiments. Error bars denote the s.e.m.