Gene transfer into nonhuman primate CD34+CD11b- bone marrow progenitor cells capable of repopulating lymphoid and myeloid lineages.
We investigated whether rhesus monkey CD34+CD11b- hematopoietic stem cells can be transduced with recombinant retroviruses carrying the human adenosine deaminase (hADA) gene by co-cultivation with a virus-producing cell line. Following autologous transplantation, polymerase chain reaction (PCR) analysis on peripheral blood mononuclear cells and granulocytes showed that the hADA-retrovirus was present in approximately 0.1% of the cells for at least 400 days post transplantation in 2 monkeys. Bone marrow that was harvested 16 months after transplantation carried ADA-overexpressing myeloid progenitor cells capable of in vitro colony formation. In addition, hADA activity could be demonstrated in T lymphocytes that were harvested 9 months post transplantation. Thus, in vitro transduction of CD34+CD11b- cells led to long-term repopulation of the hematopoietic system with transduced cells of lymphoid and myeloid lineages expressing the hADA gene. To investigate whether infusion of virus-producing cells into a rhesus monkey undergoing autologous bone marrow transplantation could lead to in vivo transfer of the recombinant retrovirus, 1 monkey was infused with CD34+CD11b- bone marrow cells (BMC) and a large quantity of virus-producing cells. Few provirus-carrying cells could temporarily be detected in this animal. This shows that in vivo gene transfer into a regenerating hemopoietic system can occur, albeit at very low efficiency.
Van Beusechem
V W
VW
Department of Gene Therapy, TNO Medical Biological Laboratory, Rijswijk, The Netherlands.
Bart-Baumeister
J A
JA
Bakx
T A
TA
Kaptein
L C
LC
Levinsky
R J
RJ
Valerio
D
D
D000243:Adenosine Deaminase / Q000235:genetics; D000818:Animals; D015703:Antigens, CD; D018952:Antigens, CD34; D018845:CD11 Antigens; D002454:Cell Differentiation; D002460:Cell Line; D004261:DNA Replication; D018014:Gene Transfer Techniques; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D008253:Macaca mulatta; D013601:T-Lymphocytes / Q000166:cytology
D015703:Antigens, CD; D018952:Antigens, CD34; D018845:CD11 Antigens; D000243:Adenosine Deaminase