TY - JOUR
T1 - Endobronchial transfection of naked viral interleukin-10 gene in rat lung allotransplantation
AU - Itano, Hideki
AU - Zhang, Wanjiang
AU - Ritter, Jon H.
AU - McCarthy, Timothy J.
AU - Yew, Nelson S.
AU - Mohanakumar, Thalachallour
AU - Patterson, G. Alexander
N1 - Funding Information:
The authors thank Paul J. Goodfellow, PhD, for assistance with gel image processing, Richard B. Schuessler, PhD, for the statistical consultation, Kathleen Grapperhaus, for experimental set-up, and Dawn Schuessler and Marry Ann Kelly for secretarial assistance. This work was supported by the National Institutes of Health grants RO1 HL-41281 (GAP) and RO1 HL56643 (TM), and Genzyme Corporation. Timothy J. McCarthy was supported by the Whitaker Foundation. Hideki Itano was supported by the Okayama University Medical School.
PY - 2001
Y1 - 2001
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0035063769&partnerID=8YFLogxK
U2 - 10.1016/S0003-4975(00)02651-5
DO - 10.1016/S0003-4975(00)02651-5
M3 - Article
C2 - 11308148
AN - SCOPUS:0035063769
SN - 0003-4975
VL - 71
SP - 1126
EP - 1133
JO - Annals of Thoracic Surgery
JF - Annals of Thoracic Surgery
IS - 4
ER -