Endobronchial transfection of naked TGF-beta1 cDNA attenuates acute lung rejection.
BACKGROUND
We investigated endobronchial transfection of CAT and TGF-beta1 cDNA selectively delivered to the lung graft with or without liposomes.


METHODS
Phase I: F344 rats received 130 microg of naked plasmid pCF1-CAT or complexed to liposome GL67 via left main bronchus instillation. Rats were awakened (pCF1-CAT, n = 4; GL67:pCF1-CAT, n = 4) or served as donors in an isogenic transplant (pCF1-CAT, n = 5; GL67:pCF1-CAT, n = 5). ELISA was performed on lungs, hearts, and livers on POD 2. Phase II: BN lungs received TGF-beta1 sense (n = 6); antisense (n = 5); GL67:TGF-beta1 sense (n = 10); or saline solution (n = 10). F344 recipients were sacrificed on POD 5. The arterial pO2 and rejection were assessed. RT-PCR for murine TGF-beta1 was performed.


RESULTS
Phase I: CAT expression was 519+/-287 pg and 63+/-68 with pCF1-CAT and 104+/-67 and 37+/-45 with GL67:pCF1-CAT, respectively, in the non-transplant and in the transplant setting. No protein was detected in the hearts, livers, and in the native lung of the recipients. Phase II: RT-PCR confirmed murine TGF-beta1 transfection. pO2 was 362.7+/-110.2 (mean mm Hg +/- SD) for sense TGF-beta1; 146.88+/-85.5 for antisense; 241.5+/-181.5 for GL67-TGF-beta1 sense; and 88.4+/-38.7 for saline. TGF-beta1 sense versus all other groups, p<0.05, GL67-TGF-beta1 sense versus saline, p = 0.01. Rejection was significantly lower for TGF-beta1 sense versus saline, p = 0.04.


CONCLUSIONS
Endobronchial administration of naked plasmid achieves selective transfection of lung grafts. Using this strategy, TGF-beta1 reduces early lung allograft rejection.
D'Ovidio
F
F
Department of Surgery, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Yano
M
M
Ritter
J H
JH
Mohanakumar
T
T
Patterson
G A
GA
D000208:Acute Disease; D000818:Animals; D001980:Bronchi; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000008:administration & dosage*; D000229:Dependovirus; D004797:Enzyme-Linked Immunosorbent Assay; D017930:Genes, Reporter; D006084:Graft Rejection / Q000628:therapy*; D008081:Liposomes; D016040:Lung Transplantation; D008297:Male; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012194:RNA-Directed DNA Polymerase; D051381:Rats; D011914:Rats, Inbred BN; D011916:Rats, Inbred F344; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000235:genetics*; D014184:Transplantation, Homologous; D014185:Transplantation, Isogeneic
D018076:DNA, Complementary; D008081:Liposomes; D016212:Transforming Growth Factor beta; D015500:Chloramphenicol O-Acetyltransferase; D012194:RNA-Directed DNA Polymerase