|
|
 Previous Article | Table of Contents | Next Article 
Retrovirus-mediated gene transduction into long-term repopulating marrow
cells of dogs
FG Schuening, K Kawahara, AD Miller, R To, S Goehle, D Stewart, K Mullally, L Fisher, TC Graham and FR Appelbaum
Transplantation Biology Program, Fred Hutchinson Cancer Research Center,
Seattle, WA 98104.
Amphotropic helper-free retrovirus vectors containing the bacterial
neomycin phosphotransferase gene (neo) and the human adenosine deaminase
gene (adenosine aminohydrolase, EC 3.5.4.4; ADA) were used to transduce
canine marrow cells. In one approach, dogs were treated for 7 days with
recombinant human granulocyte colony-stimulating factor to stimulate
hematopoietic cell division. Bone marrow cells were collected and
transduced by 24 hours of cocultivation on vector-producing cells followed
by incubation in a vector-containing long-term marrow culture system for 4
days. Transduced autologous marrow (0.4 to 1.0 x 10(8) cells/kg) was
infused into dogs administered otherwise lethal total body irradiation
(TBI) of 920 cGy. Two of four dogs engrafted, and their marrows showed
intermittently between 1% and 11% G418-resistant colony-forming unit
granulocyte-macrophage (CFU-GM) colonies for up to 2 years after
transplantation. In a different experimental approach, autologous marrow,
obtained at the time of the PB neutrophil nadir 7 days after a single
cyclophosphamide injection (40 mg/kg intravenously), was cocultivated for
24 hours on vector-producing cells and infused at doses of 0.06 to 0.18 x
10(8) cells/kg into dogs administered 920 cGy TBI. One of three dogs
engrafted, and the marrow showed intermittently 1% to 10% G418-resistant
CFU-GM colonies for at least 2 years. Culture results were confirmed by
polymerase chain reaction (PCR) showing the presence of the neo gene in
marrow cells, peripheral blood (PB) granulocytes, and PB and lymph node
lymphocytes. Dilution experiments indicated that up to 10% of marrow, lymph
node, and PB cells contained the neo gene, consistent with the culture
results. Samples harboring the neo gene also contained the gene for human
ADA. However, repeated analyses of PB and marrow cells for human ADA gene
expression by starch gel electrophoresis were negative. PB samples of all
dogs were free of helper virus, and no long-term side effects from the
transduction were observed.
Volume 78,
Issue 10,
pp. 2568-2576,
11/15/1991
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:
|
|
|
|
 
S. Barrette, J. L. Douglas, N. E. Seidel, and D. M. Bodine
Lentivirus-based vectors transduce mouse hematopoietic stem cells with similar efficiency to Moloney murine leukemia virus-based vectors
Blood,
November 15, 2000;
96(10):
3385 - 3391.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Goerner, B. Bruno, P. A. McSweeney, G. Buron, R. Storb, and H.-P. Kiem
The Use of Granulocyte Colony-Stimulating Factor During Retroviral Transduction on Fibronectin Fragment CH-296 Enhances Gene Transfer Into Hematopoietic Repopulating Cells in Dogs
Blood,
October 1, 1999;
94(7):
2287 - 2292.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Whitwam, M. E. Haskins, P. S. Henthorn, J. N. Kraszewski, S. E. Kleiman, N. E. Seidel, D. M. Bodine, and J. M. Puck
Retroviral Marking of Canine Bone Marrow: Long-Term, High-Level Expression of Human Interleukin-2 Receptor Common Gamma Chain in Canine Lymphocytes
Blood,
September 1, 1998;
92(5):
1565 - 1575.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M.K. Brenner, H.E. Heslop, D. Rill, C. Li, T. Nilson, M. Roberts, C. Smith, R. Krance, and C. Rooney
Gene Transfer and Bone Marrow Transplantation
Cold Spring Harb Symp Quant Biol,
January 1, 1994;
59(0):
691 - 697.
[Abstract]
[PDF]
|
|
|
|
|
|
|
R. Mulligan
The basic science of gene therapy
Science,
May 14, 1993;
260(5110):
926 - 932.
[Abstract]
[PDF]
|
|
|
|
|
