Fas ligand gene transfer combined with low dose cyclosporine A reduces acute lung allograft rejection.
The interaction between Fas and its ligand (FasL) induces apoptosis in the Fas-expressing cell. We hypothesized that liposome-mediated FasL gene transduction to the lung allograft, in addition to low-dose immunosuppression, might reduce acute rejection. Orthotopic left lung allotransplantation was performed in male rats (Brown Norway to Fischer F344). FasL gene transfer was performed by use of the plasmid pBCMGSNeo carrying the gene coding for murine FasL and the cationic liposome GL#67:DOPE. Six hundred and sixty micrograms of DNA in 250 microl H2O and 0.5 micromol GL#67 in 250 microl H2O were diluted to 5 ml with saline solution. This emulsion (20 degrees C) was instilled retrogradely through the left pulmonary vein after flushing with LPD solution (20 ml, at 4 degrees C). Subsequently, the graft was stored at 10 degrees C for 3 h. A single dose of cyclosporine A (CsA; 2.5 mg/kg i.m.) was given to all groups 48 h after the transplantation. In group 1 (n = 6), FasL/GL#67 was instilled as described. In group 2 (n = 5), GL#67 was given without DNA. Group 3 (n = 5) animals received CsA only. Five days after transplantation, gas exchange was assessed after exclusion of the contralateral native lung (FiO2 = 1.0). Grafts were flushed with saline solution and fixed in formaldehyde for histological evaluation. No statistical difference in gas exchange (PaO2) between the two control groups 2 (6.4 +/- 0.4 kPa) and 3 (7.4 +/- 0.4 kPa) could be detected 5 days postoperatively (P = 0.9). In contrast, grafts transduced with FasL (group 1) had significantly better gas exchange on postoperative day 5 (PaO2: group 1 37.0 +/- 10.6 kPa vs group 2 6.4 +/- 0.41 kPa; P = 0.002). Two animals in group 1 revealed no or only minimal improvement in gas exchange. Histologically, all lung specimen of all groups showed signs of acute rejection (A2). Leukocyte infiltrates, rated by two independent observers, were less severe in all group 1 animals. Liposome-mediated FasL gene transfer at the time of harvest in combination with low-dose CsA reduces acute rejection in four out of six animals in this model of rat lung allotransplantation.
Schmid
R A
RA
Department of Medicine, University Hospital Zurich, Switzerland.
Stammberger
U
U
Hillinger
S
S
Gaspert
A
A
Boasquevisque
C H
CH
Malipiero
U
U
Fontana
A
A
Weder
W
W
D000818:Animals; D016572:Cyclosporine / Q000627:therapeutic use*; D004305:Dose-Response Relationship, Drug; D053222:Fas Ligand Protein; D018014:Gene Transfer Techniques; D006084:Graft Rejection / Q000517:prevention & control*; D007165:Immunosuppression Therapy / Q000379:methods; D008081:Liposomes; D016040:Lung Transplantation / Q000276:immunology* / Q000502:physiology; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000502:physiology*; D010100:Oxygen / Q000097:blood; D010313:Partial Pressure; D010957:Plasmids; D011667:Pulmonary Veins; D051381:Rats; D011914:Rats, Inbred BN; D011916:Rats, Inbred F344; D014184:Transplantation, Homologous; D019014:fas Receptor / Q000502:physiology
D053222:Fas Ligand Protein; C505662:Faslg protein, rat; D008081:Liposomes; D008562:Membrane Glycoproteins; D019014:fas Receptor; D016572:Cyclosporine; D010100:Oxygen