Factors affecting the transduction of pluripotent hematopoietic stem cells: Long-term expression of a human adenosine deaminase gene in mice
article
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
Topics
Adenosine deaminaseInterleukin 1alphaInterleukin 3Animal cellBone marrow cellCell transplantationColony forming unit sControlled studyGene expressionGene transferGenetic transductionHematopoietic stem cellHumanMouseNonhumanRetrovirusAdenosine DeaminaseAnimalBone MarrowBone Marrow CellsBone Marrow TransplantationCells, CulturedFluorouracilGene ExpressionGenetic VectorsHematopoietic Cell Growth FactorsHematopoietic Stem CellsInterleukin-1Interleukin-3MiceMice, Inbred C57BLMice, Inbred CBARetroviridaeSpleenStem Cell FactorTransfection
TNO Identifier
232111
ISSN
00064971
Source
Blood, 81(1), pp. 254-263.
Pages
254-263
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