NBS1 is required for efficient HIV-1 transduction of primary cells. (A) NBS1-deficient fibroblasts (GM07166) and matched controls (GM04506) were infected with HIV-1-based vector carrying the lacZ reporter at an m.o.i. of 0.25. Five and seven days post infection (dpi) cells were stained using a β-galactosidase assay (Stratagene protocol) and transduced (blue) cells were counted under a light microscope the following day. Light grey – NBS1-deficient cells; dark grey – normal cells. The error bars represent standard deviation, p = 0.029 for 5 dpi and 0.021 for 7 dpi. (B) Light microscopic images from the same experiment as in A. (C) The effect of the NBS1 deficiency on expression of the lacZ marker. The NBS1-deficient and control cells were transfected with the lacZ plasmid and lacZ-expressing cells were counted three days post transfection. Six randomly selected fields were counted per each point. The error bars represent standard deviation. The differences were not statistically significant (p > 0.2) (D) Transduction with the HIV-1-based vector carrying the EGFP marker. NBS1-deficient and control fibroblasts were infected with the vector and transduced cells were counted by flow cytometry at multiple time points (2–7 dpi). Results from 7 dpi are shown. Histograms of mock infected cells (top) and cells infected at an m.o.i. of 0.1 (bottom) are shown. As seen in the gated regions, 13.45% of control fibroblasts were stably transduced, whereas transduction of NBS fibroblasts was only 4.79%.
Smith et al. Virology Journal 2008 5:11 doi:10.1186/1743-422X-5-11