1995 Fiscal Year Final Research Report Summary
Restoration of superoxide generating ability to the cells derived from chronic granulomatous disease by protein or gene transfer.
| Project/Area Number |
05557020
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Bacteriology (including Mycology)
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| Research Institution | The University of Tokyo |
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Principal Investigator |
KANEGASAKI Shiro Inst.of Med.Sci., Univ.of Tokyo Professor, 医科学研究所, 教授 (10012767)
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| Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI Sonoko Inst.of Med.Sci., Univ.of Tokyo Reserach Associate, 医科学研究所, 教務職員 (00013764)
NUNOI Hiroyuki Inst.of Med.Sci., Univ.of Tokyo Joshu, 医科学研究所, 助手 (50218260)
IMAJOH-OHMI Shinobu Inst.of Med.Sci., Univ.of Tokyo Associate Professor, 医科学研究所, 助教授 (20160046)
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| Project Period (FY) |
1993 – 1995
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| Keywords | Super oxide / CGD / Neutrophils / p67-phox / B lymphocytes / p47-phox / Vector / Immunocompromised |
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| Research Abstract |
Chronic granulomatous disease (CGD) is an inherited disorder where phagocytes and B lymphocytes cannot generate superoxide anion. Two cytosolic proteins, namely p47- and p67-phox besides cytochrome b_<558> in plasma membrane, are essential for superoxide-generation and any defect in these proteins causes CGD.By typing of 90 CGD patents in Japan, we found 9 and patients, respectively with p47- and p67-phox deficiency. GT-dinucleotide deletion (in all p47phox deficiency examined), AT dinucleotide insertion and two exon skipping (both in p67-phox deficiency) were found. We established B cell lines from these patients. Using recombinant proteins as standards, a single neutrophils was found to possess about 2.6 x 10^6 and 8.6 x 10^5 molecules of p47- and p67-phox, respectively. Approximately 4.7 x 10^5 and 3.3 x 10^4 molecules of p47- and p67-phox, respectively are present in a single peripheral B lymphocyte. We tryed in vain to restore superoxide generating ability by transfer recombinant p67-phox in liposome to a B cell line from a CGD patient. However, we could successfully constructed a retrovirus vector containing p67-phox DNA,which restored superoxide generating ability of a p67-phox deficient B cell line. Development in future : By collaborating with Dr.Y.Sugimoto, we are now constructing a bicistronic retrovirus vector that carries the human multidrug-resistance gene, MDR1 and the therapeutic gene for p67- of p47-phox deficiency on the Havey murine sarcoma retrovirus vector. The drug-selectable bicistronic vector would give higher therapeutic benefit on gene therapy of CGD patients, since retrovirus promoters cease to function after a periods.
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