Figure 2

Efficiency of transduction of cynomologus macaque primitive hematopoietic cells with SIV-based lentiviral vectors. A: Non transduced cells were used as a control for each animal. B: Transduction of bone marrow progenitor cells with an SIV-based vector. CD34⁺ cells were cultured in the presence of cytokines (see materials and methods) and exposed to vector particles at an MOI of 100 for 24 hours before FACS analysis for eGFP production. C: CD34⁺ cells were cultured overnight in a proliferation medium supplemented with various concentrations of AZT (100 nM, 1 mM, 10 mM). Cells were then washed twice and transduced with various multiplicities of infection (MOI) of the lentiviral vector (0, 1, 10, 100). After 24 hours of coculture with lentiviral vector, some of the CD34⁺ cells were used to evaluate the rate of transduction of undifferentiated CD34⁺ cells (C); * indicate statistically significant differences (Kruskal Wallis test) between cultures with and without AZT treatment for MOI = 1 (p = 0,0378), MOI = 10 (p = 0,0224) and MOI = 100 (p = 0,0247). Some of the cells were cultured for 14 days, to allow the myeloid differentiation of CFC. Cells were then resuspended, washed and fixed for three days. They were analyzed by flow cytometry, to evaluate the percentage of eGFP-positive cells and determine the rate of transduction (D); * indicates a statistically significant difference (p = 0,0237(Kruskal Wallis test)) between cultures with and without AZT treatment for MOI = 100. The results shown are the mean values for the three monkeys, each studied in triplicate.