Retroviral marking and transplantation of rhesus hematopoietic cells by nonmyeloablative conditioning.
The ability to engraft significant numbers of genetically modified hematopoietic stem and progenitor cells without the requirement for fully myeloablative conditioning therapy is a highly desirable goal for the treatment of many nonmalignant hematologic disorders. The aims of this study were to examine, in nonhuman primates (rhesus), (1) the effects of pretreatment of host animals with cytokines (G-CSF and SCF), i.e., before nonmyeloablative irradiation, on the degree and duration of neo gene marking of circulating leukocytes after autologous cell reinfusion and (2) to compare transduction of primitive hematopoietic target cells in the presence of our standard transduction cytokine combination of IL-3, IL-6, and stem cell factor (SCF) and in the presence of an alternative combination containing SCF, G-CSF, and the thrombopoietin analog MGDF. Cytokine-mobilized rhesus peripheral blood progenitor/stem cells (PBSCs) were enriched for CD34+ cells and transduced with neo vectors (either G1Na or LNL6) for 96 hr in cultures containing rhIL-3, rhIL-6, and rhSCF or MGDF, rhSCF, and rhG-CSF and cryopreserved. Four animals underwent minimal myeloablative conditioning with 500 cGy irradiation with or without pretreatment with SCF and G-CSF, followed by reinfusion of the cryopreserved cells on the subsequent day. Neutrophil nadirs (< or =500/mm3) were 0-3 days in duration; there were no significant periods of severe thrombocytopenia. Marking of circulating granulocytes and mononuclear cells was extensive and durable in all animals (exceeding 12% in the mononuclear cells of one animal) and persisted beyond the final sampling time in all animals (up to 33 weeks). No difference in extent or duration of marking was attributable to either cytokine presensitization of recipients prior to irradiation, or to the substitution of MDGF and G-CSF for IL-3 and IL-6 during transduction.
Huhn
R D
RD
Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Tisdale
J F
JF
Agricola
B
B
Metzger
M E
ME
Donahue
R E
RE
Dunbar
C E
CE
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D018952:Antigens, CD34 / Q000378:metabolism; D004351:Drug Resistance / Q000235:genetics; D018014:Gene Transfer Techniques; D005819:Genetic Markers; D005822:Genetic Vectors; D006085:Graft Survival / Q000187:drug effects* / Q000528:radiation effects*; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D006098:Granulocytes / Q000276:immunology; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000528:radiation effects / Q000821:virology; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008253:Macaca mulatta; D009355:Neomycin / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011829:Radiation Dosage; D012190:Retroviridae / Q000235:genetics*; D019089:Stem Cell Factor / Q000494:pharmacology; D014161:Transduction, Genetic; D019172:Transplantation Conditioning
D000900:Anti-Bacterial Agents; D018952:Antigens, CD34; D005819:Genetic Markers; D019089:Stem Cell Factor; D016179:Granulocyte Colony-Stimulating Factor; D009355:Neomycin