| Project/Area Number |
09671105
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Hematology
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| Research Institution | Nagoya University |
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Principal Investigator |
TANIMOTO Mitsune School of Medicine, Nagoya University Assistant Professor, 医学部, 講師 (10240805)
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| Co-Investigator(Kenkyū-buntansha) |
IIJIMA Narikazu School of Medicine, Medical Staff, 医学部, 医員
KOUNO Akio School of Medicine, Medical Staff, 医学部, 医員
唐渡 雅行 名古屋大学, 医学部, 医員
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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 |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1998: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1997: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | gene therapy / adeno associate virus / gene transduction / hematopoetic cell / cell therapy / cell culture / GVL effect / hematopoietic neoplasms / 造血幹細胞移植 / GLV効果 / ドナーリンパ球輸注 / リコンビナント / AAV |
|---|
| Research Abstract |
Adeno associated virus (AAV) has been thought to be the promising vector for gene therapy because of its wide range of human tissues and low toxicity. We have recently initiated the study of rAAV for hematopoietic cells. Preliminary results revealed that the transduction efficiencies assessed by colony formation assay employing NeoR gene into various cell lines, were CML->AML->ALL-derived cell lines by log differences. Present study was performed using peripheral T cell as a target tissue. After purification and concentration of rAAV, 27% efficiency was observed in CML line at an MOI of 7 and a linear relationship between MOI and efficiency was confirmed, suggesting that this syatem may be useful for gene transduction into leukemia cells. Integration of the NeoR gene into the host genome was assessed by Southern blotting, that revealed various sizes of DNA fragments. A fluorescent in situ hybridization (FISH) analysis was perfomed on 11 clones, in all of which the NeoR gene was integrated out of chromosome 19. In five clones, chromosome painting probes showed that the integration sites were chromosome 1q, 2q, 2q, 11p, 12p and l3q.
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