J Korean Soc Transplant.
2000 Jun;14(1):31-40.
Effect of Adenovirus Mediated Rat Interleukin-10 (Ad:ratIL-10) Gene Transfer in Mouse-to-rat Skin Graft
- Affiliations
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- 1Department of Surgery, Keimyung University School of Medicine, Taegu, Korea.
- 2Department of Pathology, Keimyung University School of Medicine, Taegu, Korea.
- 3The Institute for Medical Science, Keimyung University School of Medicine, Taegu, Korea.
Abstract
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PURPOSE: IL-10, one of the potent Th2 cytokine, has strong anti-inflammatory reaction and immunosuppression action by stabilization of polarized Th2 cell gene expression and blocking of function of antigen presenting cells. The aims of this study were to investigate the immunosuppressive effect of recombinant adenovirus mediated rat IL-10 (Ad:ratIL-10) in mouse to rat skin graft.
METHODS
The transgene expression of the recombinant adenovirus was confirmed by X-gal staining of Ad:LacZ infected skin graft. The skin graft was done between mouse and rat after 1 hour infection of donated skin with Ad:ratIL-10, which was already been produced in our Institute for Medical Science. Checked gene expression in grafted skin by in situ RT-PCR and systemic blood by ELISA on day 1, 3, 5 and 7. Immunosuppressive effect of the Ad:ratIL-10 was evaluated by graft survival and compared with control group which was infected by saline.
RESULTS
Transgene expression of recombinant adenovirus was peak on third day of skin graft and became disappeared on day 5 and 7. The same expression was confirmed by in situ RT-PCR of Ad:ratIL-10 infected skin graft. The systemic blood level of infected Ad:ratIL-10 checked by ELISA was undetectable but their expression checked in culture cell line of HeLa cell was 275 ng/mL on day 5. Mean grafted skin survival was 6.0 +/- 0.7 days in Ad:ratIL-10 group but statistically indistinguishable to control group of infection with saline (5.6 +/- 0.6 days, p>0.05).
CONCLUSION
In summary, the Ad:ratIL-10 alone, infected in grafted skin, was peak on day 3 after graft but showed no immunosuppressive effect on mouse-to-rat skin graft.