| Project/Area Number |
11671326
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Thoracic surgery
|
|---|
| Research Institution | YAMAGUCHI UNIVERSITY |
|---|
Principal Investigator |
HAMANO Kimikazu YAMAGUCHI UNIVERSITY SCHOOL OF MEDICINE, ASSISTANT PROFESSOR, 医学部, 助手 (60263787)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
GOHRA Hidenori YAMAGUCHI UNIVERSITY HOSPITAL, ASSISTANT PROFESSOR, 医学部・附属病院, 助手 (30301310)
KAJIWARA Koji YAMAGUCHI UNIVERSITY HOSPITAL, ASSISTANT PROFESSOR, 医学部・附属病院, 助手 (90253161)
FUJIMURA Yoshihiko YAMAGUCHI UNIVERSITY HOSPITAL, LECTURER, 医学部・附属病院, 講師 (60199362)
|
|---|
| Project Period (FY) |
1999 – 2000
|
| Project Status |
Completed (Fiscal Year 2000)
|
| Budget Amount *help |
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2000: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1999: ¥2,200,000 (Direct Cost: ¥2,200,000)
|
|---|
| Keywords | CTLA4-Ig / fetal cardiomyocytes / gene transfer / xenograft / artial natriuretic peptide (ANP) / infection / survive / adenovirus vectors / 細胞移植 / 心筋梗塞 / 遺伝子治療 / モノクローナル抗体 |
|---|
| Research Abstract |
Background. Experimental allografting of fetal cardiomyocytes has been performed successfully. In this study, we attempted to transplant rat fetal cardiomyocytes into the hearts of mice by blocking the CD28/B7 co-stimulatory pathway via CTLA4-Ig gene transfer.
Methods. Fetal cardiomyocytes derived from Dark Agouti (DA) rat were infected with CTLA4-Ig-expressing adenovirus vectors (AdCTLA) and injected directly into the normal myocardium of C3H/He mice (n=15). For control, cells infected with β-Gal-expressing adenovirus vector (AdRL) and cells without infection were injected into additional mice (n=30). The percentages of cardiomyocytes infection by AdRL or AdCTLA were estimated by X-gal staining or immunostaining with anti-CTLA4-Ig antibody. Mice were killed at 2 (n=5), 4 (n=5), and 8 (n=5) weeks after xenotransplantation. Transplanted fetal cardiomyocytes were examined for survival by immunostaining with anti-rat artial natriuretic peptide (ANP) and anti-CTLA4-Ig antibodies.
Results. X-gal staining and immunostaining showed infection to be approximately 30% at 10 MOI, 65% at MOI 50, and 50〜70% at MOI>100, 3 days after infection with AdCTLA or AdRL.The cardiomyocytes infected with AdCTLA expressed CTLA4-Ig and survived to 8 weeks after xenotransplantation in all of these mice. However, cardiomyocytes infected with AdRL and non-infected cells were not detected even 2 weeks after xenotransplantation.
Conclusion. Fetal cardiomyocytes were successfully infected by AdCTLA and AdRL.Survival of xenografted fetal cardiomyocytes is prolonged by adenovirus-mediated CTLA4-Ig expression.
|
