Drug-selected complete restoration of superoxide generation in Epstein-Barr virus-transformed B cells from p47
phox
-deficient chronic granulomatous disease patients by using a bicistronic retrovirus vector encoding a human multi-drug resistance gene (MDR1) and the p47
phox
gene |
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Authors: | Mayumi Iwata Hiroyuki Nunoi Ichiro Matsuda Shiro Kanegasaki Takashi Tsuruo Y Sugimoto |
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Institution: | (1) Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, 1-37-1 Kami-ikebukuro, Toshima-ku, Tokyo 170, Japan e-mail: yoshi-s@ims.u-tokyo.ac.jp, Tel.: +81-3-3918-0111, Fax: +81-3-3918-3716, JP;(2) Department of Pediatrics, Kumamoto University School of Medicine, 1-1-1 Honjo, Kumamoto 860, Japan, JP;(3) Department of Bacterial Infection, Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108, Japan, JP;(4) Institute of Molecular and Cellular Biosciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113, Japan, JP |
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Abstract: | Chronic granulomatous disease (CGD) is a group of disorders characterized by the failure of phagocytes to produce superoxide.
One-third of the cases of CGD in the USA and Europe results from defects in the gene encoding p47
phox
, a cytoplasmic component of NADPH oxidase for superoxide generation. In this study, we constructed the bicistronic retrovirus
vector Ha-MDR-IRES-p47, which carries cDNAs for a human multi-drug-resistance gene (MDR1) and p47
phox
. The amphotropic retroviral producer cells were generated, and the supernatant of the producer cells was used to transduce
Epstein-Barr virus-transformed B (EBV-B) cells, established from B cells of p47
phox
-deficient CGD patients, as an in vitro model of gene therapy for p47
phox
-deficient CGD. The transduced cells expressed both P-glycoprotein and p47
phox
protein, and the expression levels were increased after appropriate vincristine selection. The levels of superoxide production
in the vincristine-selected cells were increased to a level similar to normal EBV-B cells. This result suggests that it is
possible to achieve 100% correction of the CGD defect in p47
phox
-deficient EBV-B cells by using the bicistronic vector. This strategy could be employed not only in vitro, but also in vivo,
in the gene therapy of a number of inherited diseases.
Received: 8 June 1998 / Accepted: 5 August 1998 |
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Keywords: | |
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