Figure 6. Transduction of long-term repopulating rhesus macaque HSCs with lentiviral vectors.

Using DNA isolated from the BM of rhesus macaque RQ4876 more than 1 year after transplant, we performed LAM-PCR and isolated 10 ISs. Demonstration of common lentiviral ISs in sorted subpopulations of the PB — T cells, B cells, and granulocytes — collected before and after BG/TMZ or BG/BCNU selection was subsequently performed by PCR using 1 vector LTR primer and 1 genomic primer spanning a region of each IS. We detected 2 common ISs, providing a clear demonstration of the transduction of HSCs contributing in the long term to the different hematopoietic lineages. DNA extracted from wbcs of a nontransplanted animal was used as a negative control. Each DNA sample was amplified using β-actin–specific primers as a positive control.