Abstract
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.
Original language | English |
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Pages (from-to) | 295-305 |
Number of pages | 11 |
Journal | Human gene therapy |
Volume | 5 |
Issue number | 3 |
DOIs | |
Publication status | Published - Mar 1994 |
Keywords
- Adenosine Deaminase/genetics
- Animals
- Antigens, CD
- Antigens, CD34
- CD11 Antigens
- Cell Differentiation
- Cell Line
- DNA Replication
- Gene Transfer Techniques
- Hematopoietic Stem Cell Transplantation
- Hematopoietic Stem Cells/cytology
- Humans
- Macaca mulatta
- T-Lymphocytes/cytology