The Journal of Experimental Medicine
Janeway's Immunobiology 7th Edition
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Journal of Experimental Medicine, Vol 172, 729-736, Copyright © 1990 by Rockefeller University Press

ARTICLES

Expression of human adenosine deaminase in mice transplanted with hemopoietic stem cells infected with amphotropic retroviruses

VW van Beusechem, A Kukler, MP Einerhand, TA Bakx, AJ van der Eb, DW van Bekkum and D Valerio
Institute of Applied Radiobiology and Immunology-TNO, Rijswijk, The Netherlands.

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.
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