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Next Article 
Blood, Vol. 96 No. 1 (July 1), 2000:
pp. 1-8
PLENARY PAPER
Many multipotential gene-marked progenitor or stem cell clones
contribute to hematopoiesis in nonhuman primates
Hyeoung Joon Kim,
John F. Tisdale,
Tong Wu,
Masaaki Takatoku,
Stephanie E. Sellers,
Philipp Zickler,
Mark E. Metzger,
Brian A. Agricola,
James D. Malley,
Ikunoshin Kato,
Robert E. Donahue,
Kevin E. Brown, and
Cynthia E. Dunbar
From the Hematology Branch, National Heart, Lung and Blood
Institute, Bethesda, MD; the Molecular and Clinical Hematology Branch,
National Institute of Diabetes and Digestive and Kidney Diseases; the
Center for Information Technology, National Institutes of Health,
Bethesda, MD; and Biotechnology Research Laboratories, Takara Shuzo,
Shiga, Japan.
Retroviral insertion site analysis was used to track the
contribution of retrovirally transduced primitive progenitors to hematopoiesis after autologous transplantation in the rhesus macaque model. CD34-enriched mobilized peripheral blood cells were transduced with retroviral marking vectors containing the neo gene and
were reinfused after total body irradiation. High-level gene transfer efficiency allowed insertion site analysis of individual myeloid and
erythroid colony-forming units (CFU) and of highly
purified B- and T-lymphoid populations in 2 animals. At multiple time
points up to 1 year after transplantation, retroviral insertion sites were identified by performing inverse polymerase chain reaction and
sequencing vector-containing CFU or more than 99% pure T- and B-cell
populations. Forty-eight unique insertion sequences were detected in
the first animal and also in the second animal, and multiple clones
contributed to hematopoiesis at 2 or more time points. Multipotential
clones contributing to myeloid and lymphoid lineages were identified.
These results support the concept that hematopoiesis in large animals
is polyclonal and that individual multipotential stem or progenitor
cells can contribute to hematopoiesis for prolonged periods. Gene
transfer to long-lived, multipotent clones is shown and is encouraging
for human gene therapy applications.
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