| Project/Area Number |
08670908
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Pediatrics
|
|---|
| Research Institution | NARA MEDICAL UNIVERSITY |
|---|
Principal Investigator |
TANAKA Ichiro NARA MEDICAL UNIVERISTY,MEDICALDEPARTMENT,ASSISTANT PROFESSOR, 医学部, 講師 (00201616)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
NAKA Hiroyuki NARA MEDICAL UNIVERISTY,MEDICALDEPARTMENT,ASSISTANT PROFESSOR, 医学部, 助手 (40281761)
|
|---|
| Project Period (FY) |
1996 – 1997
|
| Project Status |
Completed (Fiscal Year 1997)
|
| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1996: ¥1,000,000 (Direct Cost: ¥1,000,000)
|
|---|
| Keywords | factor VIII / subunit / gene therapy / 遺伝子発現 / ヒト第四因子 / イヌ第四因子 / 血友病犬 / cross-reactivity |
|---|
| Research Abstract |
Each of approximately 2kb of cDNA,corresponding to factor VIII heavy and light chain, was individually inserted into the expression vector pNeoIG502. After CHO (Chinese Hamster Ovary) cells were transfected with the constracted vectors, cell supernatants were tested for the presence of factor VIII coagulant activity using both clotting and chromogenic assays, and factor VIII antigen using subunit-specific ELISAs. In the supernatant 0.02-0.04U/ml of light chain subunit was detected, whereas no heacy chain subunit was detected using subunit-specific ELISAs. Factor VIII coagulant activity was also undetectable by both clotting and chromogenic assays. When adenoviral vector was used as the expression vector instead of pNeoIG502,0.1-0.2U/ml of light chain subunit was detected in the supernatants of cells with no detection of heavy chain and coagulant activity of factor VIII.
Further investigations have been attemptted using various vectors, such as retroviral or adeno-associated viral vector as the expression vector, and hepatoma cells (HepG2), fibroblast cells, or myoblast cells as the target cell.
|
