title: Immunotargeting of glucose oxidase to endothelium in vivo causes oxidative vascular injury in the lungs.
abstract: Vascular immunotargeting is a novel approach for site-selective drug delivery to endothelium. To validate the strategy, we conjugated glucose oxidase (GOX) via streptavidin with antibodies to the endothelial cell surface antigen platelet endothelial cell adhesion molecule (PECAM). Previous work documented that 1) anti-PECAM-streptavidin carrier accumulates in the lungs after intravenous injection in animals and 2) anti-PECAM-GOX binds to, enters, and kills endothelium via intracellular H(2)O(2) generation in cell culture. In the present work, we studied the targeting and effect of anti-PECAM-GOX in animals. Anti-PECAM-GOX, but not IgG-GOX, accumulated in the isolated rat lungs, produced H(2)O(2,) and caused endothelial injury manifested by a fourfold elevation of angiotensin-converting enzyme activity in the perfusate. In intact mice, anti-PECAM-GOX accumulated in the lungs (27 +/- 9 vs. 2.4 +/- 0.3% injected dose/g for IgG-GOX) and caused severe lung injury and 95% lethality within hours after intravenous injection. Endothelial disruption and blebbing, elevated lung wet-to-dry ratio, and interstitial and alveolar edema indicated that anti-PECAM-GOX damaged pulmonary endothelium. The vascular injury in the lungs was associated with positive immunostaining for iPF(2alpha)-III isoprostane, a marker for oxidative stress. In contrast, IgG-GOX caused a minor lung injury and little (5%) lethality. Anti-PECAM conjugated with inert proteins induced no death or lung injury. None of the conjugates caused major injury to other internal organs. These results indicate that an immunotargeting strategy can deliver an active enzyme to selected target cells in intact animals. Anti-PECAM-GOX provides a novel model of oxidative injury to the pulmonary endothelium in vivo.
authors.affiliation: Pulmonary Critical Care Division, Department of Medicine, University of Pennsylvania Medical Center, Philadelphia 19104, USA.
authors.forename: M
authors.initials: M
authors.lastname: Christofidou-Solomidou
authors.forename: G G
authors.initials: GG
authors.lastname: Pietra
authors.forename: C C
authors.initials: CC
authors.lastname: Solomides
authors.forename: E
authors.initials: E
authors.lastname: Arguiris
authors.forename: D
authors.initials: D
authors.lastname: Harshaw
authors.forename: G A
authors.initials: GA
authors.lastname: Fitzgerald
authors.forename: S M
authors.initials: SM
authors.lastname: Albelda
authors.forename: V R
authors.initials: VR
authors.lastname: Muzykantov
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000201:enzymology*; D018390:Gene Targeting; D005949:Glucose Oxidase / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D066298:In Vitro Techniques; D008171:Lung Diseases / Q000139:chemically induced / Q000473:pathology; D051379:Mice; D018384:Oxidative Stress; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000276:immunology; D011652:Pulmonary Circulation; D051381:Rats; D014018:Tissue Distribution; D014652:Vascular Diseases / Q000139:chemically induced* / Q000473:pathology
chemical_list: D000906:Antibodies; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D005949:Glucose Oxidase