Figure 2.

Ectopic expression of GATA-1 potently reduces cell-surface expression of CCR5 in primary human cell targets of HIV-1. (A) Primary human HSC-derived DCs were transduced with HDV, HDV.GATA-1, or HDV.GATA-3 and cultured for an additional 4 days. CCR5 expression levels on HSC-derived DCs were determined via flow cytometric analyses by costaining cells with anti-CCR5, anti-CD1c (Miltenyi Biotech) to monitor DC differentiation and anti-mCD24 to resolve DCs expressing the lentiviral transgene. (B) Primary human conventional T-cell subsets (top 3 panels) or NKT cells (bottom) were TCR activated and transduced to express GATA-1, GATA-3, or the control HDV (see “Materials and methods”). Cells were expanded in the presence of IL-2, and cell-surface CCR5 expression was determined on unsorted populations of transduced cells and costained with anti-mCD24 antibodies. Additionally, NKT cells were costained with a fluorescein isothiocyanate (FITC)–conjugated anti–human Vβ11 antibody (Coulter), to exclude any contaminating non-NKT cells. These results are representative of at least 3 experiments performed on each cell type from unique adult or umbilical cord blood preparations.