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Right arrow Lung - transplantation

Ann Thorac Surg 2001;71:1126-1133
© 2001 The Society of Thoracic Surgeons

Original article: general thoracic

Endobronchial transfection of naked viral interleukin-10 gene in rat lung allotransplantation

Hideki Itano, MDa, Wanjiang Zhang, MDa, Jon H. Ritter, MDb, Timothy J. McCarthy, PhDc, Nelson S. Yew, PhDd, Thalachallour Mohanakumar, PhDa, G. Alexander Patterson, MDa

a Division of Cardiothoracic Surgery, Washington University School of Medicine, St. Louis, Missouri, USA
b Department of Pathology, Washington University School of Medicine, St. Louis, Missouri, USA
c Department of Radiology, Washington University School of Medicine, St. Louis, Missouri, USA
d Genzyme Corporation, Framingham, Massachusetts, USA

Address reprint requests to Dr Patterson, Division of Cardiothoracic Surgery, Washington University School of Medicine, 3108 Queeny Tower, One Barnes-Jewish Hospital Plaza, St. Louis, MO 63110
e-mail: kellym{at}msnotes.wustl.edu

Presented at the Poster Session of the Thirty-sixth Annual Meeting of The Society of Thoracic Surgeons, Fort Lauderdale, FL, Jan 31–Feb 2, 2000.

Background. Recent studies suggest that viral interleukin-10 (vIL-10) suppresses alloimmune response in transplantation. Tissue mRNA expression of inducible nitric oxide synthase (iNOS) and exhaled nitric oxide (NO) levels have been observed to increase in lung allograft rejection. The aims of this study were to examine the feasibility of vIL-10 gene transfer into rat lung allografts and to investigate its effect on subsequent allograft rejection.

Methods. Male Lewis rats (RT1l) underwent left lung transplantation with allografts from Brown Norway rats (RT1n). The donor rats were endobronchially transfected 2 minutes before harvest with 400 µg (group I, n = 5), 600 µg (group II, n = 5), or 800 µg (group III, n = 5) of naked pCMVievIL-10. Group IV (n = 5) animals, serving as control, received 400 µg of naked pCF1-CAT. All recipients were sacrificed on postoperative day 5. Transgene expression of vIL-10 was assessed by both reverse transcriptase-polymerase chain reaction and immunohistochemistry. Allograft gas exchange, exhaled NO level, histologic rejection score, and mRNA expression of graft cyokines were also assessed.

Results. Transgene expression of lung graft vIL-10 was detected by both reverse transcriptase-polymerase chain reaction and immunohistochemistry. The iNOS mRNA expression in groups I, II, and III was significantly lower than that of group IV (p < 0.05, analysis of variance). Exhaled NO levels in groups I, II, and III were significantly lower than in group IV (p < 0.01, analysis of variance). There was no significant difference between groups with respect to gas exchange, peak airway pressure, or histologic rejection score.

Conclusions. It appears that endobronchial transfection of naked vIL-10 plasmid in a rat lung allotransplant model is feasible and suppresses lung iNOS mRNA expression and exhaled NO levels. An association between iNOS upregulation and high exhaled NO levels in lung allograft resection was also noted.




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