Protection of bone marrow transplant recipients from lethal doses of methotrexate by the generation of methotrexate-resistant bone marrow.
To develop a highly efficient means for generating methotrexate resistant (MTXr) hematopoietic cells in vivo, a recombinant retroviral genome was constructed that encodes a MTXr dihydrofolate reductase (DHFRr). Cell lines producing high titers of virus capable of transmitting the DHFR gene were generated and used to infect mammalian cells in vitro. Analysis of infected fibroblasts indicated that the DHFRr gene was transmitted intact and conferred a high level of MTXr upon cells. Based on these findings, DHFRr-containing virus was used to infect murine bone marrow cells in vitro. Following infection, the transduced cells were introduced into lethally irradiated recipients via bone marrow transplantation techniques. The presence of the proviral sequences in cells of the spleen and bone marrow of engrafted recipients was associated with significantly increased survival of mice treated with otherwise lethal doses of MTX.
15 references, click to expand
-
The growth of mouse bone marrow cells in vitro.Aust J Exp Biol Med Sci. 1966 Jun;44(3):287-99 PMID: 4164182
-
Human purine nucleoside phosphorylase and adenosine deaminase: gene transfer into cultured cells and murine hematopoietic stem cells by using recombinant amphotropic retroviruses.Mol Cell Biol. 1987 Feb;7(2):838-46 PMID: 3102947
-
Evidence for early recruitment of granulocyte precursors during high-dose methotrexate infusions in mice.Blood. 1976 Aug;48(2):301-7 PMID: 949552
-
Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter.J Mol Appl Genet. 1982;1(4):327-41 PMID: 6286831
-
Isolation and expression of an altered mouse dihydrofolate reductase cDNA.Proc Natl Acad Sci U S A. 1983 May;80(9):2495-9 PMID: 6573667
-
Construction and applications of a highly transmissible murine retrovirus shuttle vector.Cell. 1984 Jul;37(3):1053-62 PMID: 6331674
-
Introduction of new genetic material into pluripotent haematopoietic stem cells of the mouse.Nature. 1984 Aug 9-15;310(5977):476-80 PMID: 6087158
-
High-efficiency gene transfer into mammalian cells: generation of helper-free recombinant retrovirus with broad mammalian host range.Proc Natl Acad Sci U S A. 1984 Oct;81(20):6349-53 PMID: 6093098
-
Introduction of a selectable gene into primitive stem cells capable of long-term reconstitution of the hemopoietic system of W/Wv mice.Cell. 1985 Aug;42(1):71-9 PMID: 4016956
-
Expression of a foreign gene in myeloid and lymphoid cells derived from multipotent haematopoietic precursors.Nature. 1985 Nov 14-20;318(6042):149-54 PMID: 3903518
-
Somatic gene therapy. Current status and future prospects.J Clin Invest. 1986 Apr;77(4):1053-6 PMID: 3514670
-
Retrovirus-mediated transfer and expression of drug resistance genes in human haematopoietic progenitor cells.Nature. 1986 Mar 20-26;320(6059):275-7 PMID: 3960109
-
Retrovirus-mediated transfer of human adenosine deaminase gene sequences into cells in culture and into murine hematopoietic cells in vivo.Proc Natl Acad Sci U S A. 1986 Apr;83(8):2566-70 PMID: 3458218
-
Developmental potential and dynamic behavior of hematopoietic stem cells.Cell. 1986 Jun 20;45(6):917-27 PMID: 2871944
-
Erythroid colony formation in cultures of mouse and human bone marrow: analysis of the requirement for erythropoietin by gel filtration and affinity chromatography on agarose-concanavalin A.J Cell Physiol. 1974 Apr;83(2):309-20 PMID: 4822532
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