Title
Factors affecting the transduction of pluripotent hematopoietic stem cells: Long-term expression of a human adenosine deaminase gene in mice
Author
Einerhand, M.P.W.
Bakx, T.A.
Kukler, A.
Valerio, D.
Radiobiologisch Instituut TNO
Publication year
1993
Abstract
An amphotropic retroviral vector, LgAL(ΔMo + PyF101) containing a human adenosine deaminase (ADA) cDNA was used to optimize procedures for the lasting genetic modification of the hematopoietic system of mice. The highest number of retrovirally infected cells in the hematopoietic tissues of long- term reconstituted mice was observed after transplantation of bone marrow (BM) cells that had been cocultured in the presence of both interleukin-1α (IL-1α) and IL-3. A significantly lower number was detected when IL-1α was omitted from such cocultures. The yield of cells that generate spleen colony- forming cells (CFU-S) in the BM of lethally irradiated recipients (MRA-CFU- S) significantly improved on inclusion of the adherent cell fraction of cocultures in the transplant. Retroviral integration patterns in MRA-CFU-S- derived spleen colonies showed that an MRA-CFU-S can produce many CFU-S during BM regeneration. Expression of hADA was detected in the circulating white blood cells of long-term reconstituted animals, demonstrating that the LgAL(ΔMo + PyF101) vector is capable of directing the sustained expression of hADA, and in approximately 35% of the transduced MRA-CFU-S-derived spleen colonies. These results should facilitate the development of gene therapy protocols for the treatment of severe combined immunodeficiency caused by a lack of functional ADA. Chemicals/CAS: adenosine deaminase, 9026-93-1; Adenosine Deaminase, EC 3.5.4.4; Fluorouracil, 51-21-8; Genetic Vectors; Hematopoietic Cell Growth Factors; Interleukin-1; Interleukin-3; Stem Cell Factor
Subject
Adenosine deaminase
Interleukin 1alpha
Interleukin 3
Animal cell
Bone marrow cell
Cell transplantation
Colony forming unit s
Controlled study
Gene expression
Gene transfer
Genetic transduction
Hematopoietic stem cell
Human
Mouse
Nonhuman
Retrovirus
Adenosine Deaminase
Animal
Bone Marrow
Bone Marrow Cells
Bone Marrow Transplantation
Cells, Cultured
Fluorouracil
Gene Expression
Genetic Vectors
Hematopoietic Cell Growth Factors
Hematopoietic Stem Cells
Interleukin-1
Interleukin-3
Mice
Mice, Inbred C57BL
Mice, Inbred CBA
Retroviridae
Spleen
Stem Cell Factor
Transfection
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TNO identifier
232111
ISSN
0006-4971
Source
Blood, 81 (1), 254-263
Document type
article