| Project/Area Number |
09557045
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
内科学一般
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| Research Institution | The University of Tokyo |
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Principal Investigator |
KANEGASAKI Shiro The Univ.of Tokyo, The Inst.of Med.Sci., Professor, 医科学研究所, 教授 (10012767)
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| Co-Investigator(Kenkyū-buntansha) |
NUNOI Hiroyuki Kumamoto University, School of Med., Associate Professor, 医学部, 助教授 (50218260)
KOBAYASHI Sonoko The Univ.of Tokyo, The Inst.of Med.Sci., Research Associate, 医科学研究所, 教務職員 (00013764)
島津 光伸 三菱BCL, 研究員
松本 良二 東京大学, 医科学研究所, 助手 (20272495)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥11,700,000 (Direct Cost: ¥11,700,000)
Fiscal Year 1998: ¥5,500,000 (Direct Cost: ¥5,500,000)
Fiscal Year 1997: ¥6,200,000 (Direct Cost: ¥6,200,000)
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| Keywords | superoxide generation / retroviral vector / chronic granulomatous disease / MDR gene / gene therapy / gp91 / p47 / p67 / 遺伝子導入 / 食細胞 / MDR / 好中球 / B細胞株 |
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| Research Abstract |
Chronic granulomatous disease is an inherited disorder where phagocytes can not generate reactive oxygen species to kill infectious agents and most of the patients with this disease die from severe infections before age 20. The disease is a result from a defect in any of the four single genes essential for superoxide generation ; namely, genes for gp91-phox and p22-phox that compose cytochrome b558 and those for p47-and p67-phox. We performed experiments toward gene therapy for CGD patients by testing two kinds of retroviral vectors for correction of patient's cells. One type of the vectors developed is bicistronic retrovirus vectors, Ha-MDR-IRES-gp91, Ha-MDR-IRES-p47 and Ha-MDR-IRES-p67 ; Ha-MDR-IRES-p22. In the construct, the MDR1 gene is translated in a cap-dependent manner and the phox genes under the control of an internal ribosome entry site(IRES). When Ha-MDR-IRES-gp91 was introduced into B cell lines derived from X-linked CGD patients or bone marrow cells from the gp91-phox knockout mice, cytochrome b558 was expressed (together with P-glycoprotein, the product of MDR gene). The cells acquired the similar level of superoxide generating activity as that of normal controls but only after selection with vincristin. The drug-selectable bicistronic vectors would give higher benefit on the gene therapy of CGD patients when we can obtain high titer particles using a stable 293 amphotropic packaging line. Using this line, he could obtaine a very high titer of MFGS-91 particles(10^7)and clinically used in US.We demonstrated that gp91-phox was efficiently expressed in B cell lines from X-linked patients when transduced useing this vector.
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