Many multipotential gene-marked progenitor or stem cell clones contribute to hematopoiesis in nonhuman primates

Retroviral Insertion site analysis was used to track the contribution of re trovirally transduced primitive progenitors to hematopoiesis after autologo us transplantation in the rhesus macaque model. CD34-enriched mobilized per ipheral blood cells were transduced with retroviral marking vectors contain ing the neo gene and were reinfused after total body irradiation. High-leve l gene transfer efficiency allowed insertion site analysis of individual my eloid and erythroid colony-forming units (CFU) and of highly purified B- an d T-lymphoid populations in 2 animals. At multiple time points up to 1 year after transplantation, retroviral insertion sites were identified by perfo rming inverse polymerase chain reaction and sequencing vector-containing CF U or more than 99% pure T- and B-cell populations. Forty-eight unique inser tion sequences were detected in the first animal and also in the second ani mal, and multiple clones contributed to hematopoiesis at 2 or more time poi nts. Multipotential clones contributing to myeloid and lymphoid lineages we re identified. These results support the concept that hematopoiesis in larg e animals is polyclonal and that individual multipotential stem or progenit or cells can contribute to hematopoiesis for prolonged periods. Gene transf er to long-lived, multipotent clones is shown and is encouraging for human gene therapy applications. (Blood. 2000; 96:1-8) (C) 2000 by The American S ociety of Hematology.