아데노바이러스를 이용한 쥐의 재조합 IL-10 (Ad:ratIL-10)의 거부반응 억제효과

Abstract

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 (의과학연구소). 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.

아데노바이러스를 vector로 한 재조합 IL-10 (Ad:ratIL-10)이 동향성 이종이식에서 거부반응 억제효과를 갖는지 알아보기 위해 ICR-mouse의 피부를 절제하여 Ad:ratIL-10에 감염시킨 후 Lewis rat에 이식하여 생착정도롤 조사하였다. 감염된 바이러스의 발현은 10^9 pfu/mL 농도에서 60분 감염시 이식 3일째에 절정을 보였고, 이식 5일 및 7일째는 다시 소멸된 것을 알 수 있었다. 쥐의 IL-10을 배양세포주에 감염시켰을 때 발현정도는 감염 후 5일째에 275 ng/mL 정도였으나, 생체에서 감염시킨 피부를 이식한 후 측정한 혈중농도는 ELISA검사에서 이식 후 어느 날에도 측 정되지 않을 정도로 소량이어서 국소감염된 Ad:ratIL-10의 전신 효과는 없음올 보여 주었다. Ad:ratIL-10 감염군의 피부생착기간은 6.0±0.7일로 대조군의 5.6 ±0.6일보다 약간 길었으나 통계적 유의성은 없었다 (p>0.05).