Title
Expression of human adenosine deaminase in mice transplanted with hemopoietic stem cells infected with amphotropic retroviruses
Author
Instituut voor Toegepaste Radiobiologie en Immunologie TNO
van Beusechem, V.W.
Kukler, A.
Einerhand, M.P.W.
Bakx, T.A.
van der Eb, A.J.
van Bekkum, D.W.
Valerio, D.
Publication year
1990
Abstract
Amphotropic recombinant retroviruses were generated carrying sequences encoding human adenosine deaminase (ADA). Transcription of the human ADA gene was under control of a hybrid long terminal repeat in which the enhancer from the Moloney murine leukemia virus was replaced by an enhancer from the F101 host-range mutant of polyoma virus. Hemopoietic stem cells in murine bone marrow were infected with this virus under defined culture conditions. As a result, 59% of day-12 colony forming unit spleen (CFU-S) stem cells became infected without any in vitro selection. Infected CFU-S were shown to express human ADA before transplantation and this expression sustained upon in vivo maturation. Mice transplanted with infected bone marrow exhibited human ADA expression in lymphoid, myeloid, and erythroid cell types. Moreover, human ADA expression persisted in secondary and tertiary transplanted recipients showing that human ADA-expressing cells were derived from pluripotent stem cells. These characteristics of our amphotropic viruses make them promising tools in gene therapy protocols for the treatment of severe combined immunodeficiency caused by ADA deficiency. In this respect it is also relevant that the viral vector that served as backbone for the ADA vector was previously shown to be nonleukemogenic.
Subject
adenosine deaminase
animal experiment
article
combined immunodeficiency
enzyme deficiency
gene expression
genetic engineering
genetic transcription
hematopoietic stem cell
human
mouse
priority journal
retrovirus
Adenosine Deaminase
Animal
Blotting, Southern
Bone Marrow Transplantation
Cell Line
Cells, Cultured
DNA
Gene Expression
Genetic Vectors
Hematopoietic Stem Cell Transplantation
Human
Mice
Nucleoside Deaminases
Recombination, Genetic
Retroviridae
Support, Non-U.S. Gov't
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DOI
https://doi.org/10.1084/jem.172.3.729
TNO identifier
231232
ISSN
0022-1007
Source
Journal of Experimental Medicine, 172 (172), 729-736
Document type
article