1991 Fiscal Year Final Research Report Summary
Development of the Methods for Regulated Expression of Transduced Gene in Transkaryotic Gene Therapy
| Project/Area Number |
01870051
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
内分泌・代謝学
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| Research Institution | The University of Tokushima (1990-1991)
University of Tsukuba (1989) |
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Principal Investigator |
ITAKURA Mitsuo Sch. of Med., Univ. of Tokushima, Prof., 医学部, 教授 (60134227)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAUCHI Hiromitsu The Inst. of Physic. and Chem. Res., Researcher, 国際フロンティア研究システム クロモゾーム研究チーム, 研究員 (40175485)
NAGATA Akihiko Sumitomo Pharmaceut. Co., Researcher, 研究所, 研究員
IWAHANA Hiroyuki Sch. of Med., Univ. of Tokushima, Res. Assist., 医学部, 助手
YOSHIMOTO Katsuhiko Sch. of Med., Univ. of Tokushima, Assoc. Prof., 医学部, 助教授 (90201863)
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| Project Period (FY) |
1989 – 1991
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| Keywords | gene therapy / diabetes mellitus / fibrablasts / proinsulin / metallothionein / differentiation antigen / monoclonal antibody / transkaryotic |
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| Research Abstract |
The principle of controlling gene expression in transkaryotic or somatic cell gene therapy was developped. The practical methods used in this study were as follows : At first recombinant human insulin cDNA constucted in the plasmid of pBMG-Neo was transfected to mouse cultured fibroblasts of L-cells. Among the clones, the one with the highest proinsulin secretion was selected. Then the second recombinant plasmid containing mouse genomic CD8.2 gene in pHEBo was further transfected to this proinsulin producing cell line. This doubly transfected cells produced proinsulin at 3.4x10^<-6> ng/hr/cell. 2x10^6 of this cell line were intraperitoneally transplanted to streptozocininduced diabetic C3H mice. The blood glucose concentrations were remarkably decreased from 430 mg/dl of the pretreatment level to 80 mg/dl at the 30th day after the transplantation. As expected, animals died of hypoglycemia due to the excessive production of proinsulin. To remove the transplanted cells, an immunological safety system using anti-CD8.2 monoclonal antibodywas tested. The administation of this antibody on the 14th day after the transplantation, once a day for 14 days, completely revered the hypoglycemic effects of transplantation, proving the complete removal of transplated cells. Thus we have developed the animal model of somatic cell gene therapy against diabetes, with the immunological safety system eligible to completely remove the transplanted cells.
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