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

Mayumi Iwata, Hiroyuki Nunoi, Ichiro Matsuda, Shiro Kanegasaki, Takashi Tsuruo, Yoshikazu Sugimoto

Research output: Contribution to journalArticle

12 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)419-423
Number of pages5
JournalHuman Genetics
Volume103
Issue number4
DOIs
Publication statusPublished - 1998
Externally publishedYes

Fingerprint

Chronic Granulomatous Disease
Multiple Drug Resistance
Retroviridae
Human Herpesvirus 4
Superoxides
B-Lymphocytes
Pharmaceutical Preparations
Genes
Vincristine
Genetic Therapy
NADPH Oxidase
P-Glycoprotein
Phagocytes
4-ethoxymethylene-2-phenyl-2-oxazoline-5-one
Complementary DNA
Proteins

ASJC Scopus subject areas

  • Genetics(clinical)
  • Genetics

Cite this

@article{1ee0fffcdc784618b0b3134d5a37d288,
title = "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",
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.",
author = "Mayumi Iwata and Hiroyuki Nunoi and Ichiro Matsuda and Shiro Kanegasaki and Takashi Tsuruo and Yoshikazu Sugimoto",
year = "1998",
doi = "10.1007/s004390050844",
language = "English",
volume = "103",
pages = "419--423",
journal = "Human Genetics",
issn = "0340-6717",
publisher = "Springer Verlag",
number = "4",

}

TY - JOUR

T1 - 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

AU - Iwata, Mayumi

AU - Nunoi, Hiroyuki

AU - Matsuda, Ichiro

AU - Kanegasaki, Shiro

AU - Tsuruo, Takashi

AU - Sugimoto, Yoshikazu

PY - 1998

Y1 - 1998

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0031782619&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031782619&partnerID=8YFLogxK

U2 - 10.1007/s004390050844

DO - 10.1007/s004390050844

M3 - Article

VL - 103

SP - 419

EP - 423

JO - Human Genetics

JF - Human Genetics

SN - 0340-6717

IS - 4

ER -