Pre-ischaemic conditioning of the pulmonary endothelium by immunotargeting of catalase via angiotensin-converting-enzyme antibodies.
BACKGROUND
Alleviation of oxidative stress via targeted delivery of catalase to the pulmonary endothelium by conjugation of angiotensin-converting-enzyme (ACE) monoclonal antibodies attenuates lung injury in an in vivo model of warm lung ischaemia and reperfusion. This study evaluates treatment of lung allografts with conjugates of anti-ACE antibody with catalase (9B9-CAT) in the setting of hypothermic preservation and reports the effect on ischaemia/reperfusion injury in this model.


METHODS
Rats were injected 1h prior to lung harvesting with mouse immunoglobulin G (IgG) (negative controls), catalase only (CAT) or anti-ACE mAb 9B9 conjugated with catalase (9B9-CAT). Lungs were flushed with low-potassium dextran (LPD) solution, excised and stored at 4 degrees C for 4 and 8h. Grafts were isolated and directly reperfused at 37 degrees C for up to 180 min. Peak inspiratory pressure (PIP), pulmonary arterial pressure (PAP) and lung weight were measured during reperfusion. Anti-oxidative capacity and catalase activity were measured in frozen lung tissue and inflammatory parameters were detected during reperfusion in perfusate solution.


RESULTS
Cold ischaemia of 8h significantly increased lung weight gain, PIP and PAP in non-immune mouse IgG and CAT-treated lungs than in 9B9-CAT-treated lungs (p<0.005). Significantly higher catalase activity and anti-oxidative status were found in the lung tissue of animals conditioned with 9B9-CAT after 4 and 8h of cold storage than in animals treated with catalase (CAT) alone or in animals treated with non-immune mouse IgG (p<0.01).


CONCLUSIONS
These results validate immunotargeting by anti-ACE mAb conjugated with catalase as a prospective and specific strategy to augment anti-oxidative defence of the pulmonary endothelium during lung transplantation. Vascular immunotargeting of anti-oxidative enzymes could limit reactive oxygen species mediated ischaemia-reperfusion (I/R) injury of the lung and has the potential to become a promising modality for extension of the viability of banked transplantation tissue.
Nowak
Kai
K
Division of Thoracic Surgery, Department of Surgery, Mannheim University Medical Center, Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68135 Mannheim, Germany. kai.nowak@chir.ma.uni-heidelberg.de
Hanusch
Christine
C
Nicksch
Kathrin
K
Metzger
Roman P
RP
Beck
Grietje
G
Gebhard
Martha M
MM
Hohenberger
Peter
P
Danilov
Sergei M
SM
D000806:Angiotensin-Converting Enzyme Inhibitors / Q000276:immunology / Q000627:therapeutic use*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000627:therapeutic use; D000975:Antioxidants / Q000378:metabolism; D002374:Catalase / Q000276:immunology* / Q000378:metabolism; D050377:Cold Ischemia / Q000379:methods*; D015925:Cryopreservation; D004730:Endothelium, Vascular / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000627:therapeutic use; D018796:Immunoconjugates / Q000627:therapeutic use; D019194:Ischemic Preconditioning / Q000379:methods*; D008168:Lung / Q000098:blood supply* / Q000378:metabolism; D008297:Male; D018384:Oxidative Stress; D011654:Pulmonary Edema / Q000378:metabolism / Q000473:pathology; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000378:metabolism / Q000517:prevention & control*
D000806:Angiotensin-Converting Enzyme Inhibitors; D000911:Antibodies, Monoclonal; D000975:Antioxidants; D004791:Enzyme Inhibitors; D018796:Immunoconjugates; D002374:Catalase