|
|
 Previous Article | Table of Contents | Next Article 
Retroviral gene transfer of human adenosine deaminase in murine
hematopoietic cells: effect of selectable marker sequences on long-term
expression
JF Apperley, BD Luskey and DA Williams
Howard Hughes Medical Institute, Department of Pediatrics, Children's
Hospital, Boston, MA 02115.
Retroviral-mediated gene transfer of human adenosine deaminase (hADA)
provides a model system for the development of somatic gene therapy as a
therapy for diseases of bone marrow-derived cells. We have previously
demonstrated that hADA can be observed in all hematopoietic lineages in a
minority of mice transplanted with bone marrow cells infected with a
simplified retroviral vector, ZipPGK-ADA. Here we report a majority of mice
(six of eight) demonstrate expression of hADA in the peripheral blood at
least 6 months after transplantation with bone marrow infected with this
simplified retroviral vector, which contains no selectable marker. The
failure to express hADA in two of eight mice was associated with the
absence of the recombinant retroviral provirus in DNA prepared from bone
marrow cells of these mice apparently due to failure to efficiently infect
the reconstituting hematopoietic stem cell. In an effort to preselect bone
marrow stem cells containing proviral integrations, we incorporated the
selectable marker neo phosphotransferase (NEO) into a retroviral vector
encoding hADA, N2/ZipPGK-ADATKNEO, and used G418 selection of infected bone
marrow cells before transplantation. In contrast to the simplified
retroviral vector, hADA expression in these recipients was short lived
(less than 8 weeks), despite the continued presence of intact provirus in
DNA prepared from bone marrow of these mice. To determine whether the
preselection of bone marrow using G418 was responsible for the lack of
sustained hADA expression, we repeated the infection with the N2/ZipPGK-
ADATKNEO vector but omitted the G418 selection step. Again, the majority of
recipient mice failed to express hADA long term, although the continued
presence of provirus in DNA prepared from peripheral blood cell mononuclear
cells was clearly demonstrated. Finally, we demonstrate clonal fluctuation
of infected stem cells, and observe a temporal correlation between
cessation of expression of hADA and the emergence of a dominant stem cell
clone between 14 and 20 weeks posttransplantation in one recipient. These
data suggest that inclusion of a second transcriptional unit that includes
neo phosphotransferase sequences in this simplified vector is associated
with decreased expression of the nonselectable ADA sequences.
Volume 78,
Issue 2,
pp. 310-317,
07/15/1991
Copyright © 1991 by The American Society of Hematology
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
T. W. Austin, S. Salimi, G. Veres, F. Morel, H. Ilves, R. Scollay, and I. Plavec
Long-term multilineage expression in peripheral blood from a Moloney murine leukemia virus vector after serial transplantation of transduced bone marrow cells
Blood,
February 1, 2000;
95(3):
829 - 836.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. A. Delviks and V. K. Pathak
Development of Murine Leukemia Virus-Based Self-Activating Vectors That Efficiently Delete the Selectable Drug Resistance Gene during Reverse Transcription
J. Virol.,
October 1, 1999;
73(10):
8837 - 8842.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K.L. MacKenzie, A. Dolnikov, M. Millington, Y. Shounan, and G. Symonds
Mutant N-ras Induces Myeloproliferative Disorders and Apoptosis in Bone Marrow Repopulated Mice
Blood,
March 15, 1999;
93(6):
2043 - 2056.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Pei, L. A. Barber, R. C. Murphy, C. A. Johnson, S. W. Kelley, L. C. Dy, R. H. Fertel, T. M. Nguyen, D. A. Williams, and J. B. Travers
Activation of the Epidermal Platelet-Activating Factor Receptor Results in Cytokine and Cyclooxygenase-2 Biosynthesis
J. Immunol.,
August 15, 1998;
161(4):
1954 - 1961.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. A. Bunnell, M. Metzger, E. Byrne, R. A. Morgan, and R. E. Donahue
Efficient In Vivo Marking of Primary CD4+ T Lymphocytes in Nonhuman Primates Using a Gibbon Ape Leukemia Virus-Derived Retroviral Vector
Blood,
March 15, 1997;
89(6):
1987 - 1995.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
