Endobronchial transfection of naked viral interleukin-10 gene in rat lung allotransplantation.
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 microg (group I, n = 5), 600 microg (group II, n = 5), or 800 microg (group III, n = 5) of naked pCMVievIL-10. Group IV (n = 5) animals, serving as control, received 400 microg 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.
Itano
H
H
Department of Pathology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Zhang
W
W
Ritter
J H
JH
McCarthy
T J
TJ
Yew
N S
NS
Mohnanakumar
T
T
Patterson
G A
GA
D000704:Analysis of Variance; D000818:Animals; D001483:Base Sequence; D001999:Bronchoscopy; D005786:Gene Expression Regulation; D006084:Graft Rejection; D006085:Graft Survival; D007150:Immunohistochemistry; D016753:Interleukin-10 / Q000235:genetics*; D016040:Lung Transplantation / Q000276:immunology* / Q000379:methods*; D008297:Male; D023421:Models, Animal; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000032:analysis; D016133:Polymerase Chain Reaction; D011336:Probability; D051381:Rats; D011914:Rats, Inbred BN; D011917:Rats, Inbred Lew; D012680:Sensitivity and Specificity; D018709:Statistics, Nonparametric; D014162:Transfection; D014181:Transplantation Immunology; D014184:Transplantation, Homologous; D014764:Viral Proteins / Q000235:genetics*
C066460:BCRF1 protein, Human herpesvirus 4; D014764:Viral Proteins; D016753:Interleukin-10; D009569:Nitric Oxide