Specific killing of human endothelial cells by antibody-conjugated glucose oxidase

1-Cys peroxiredoxin (1-cysPrx) is a novel antioxidant enzyme able to reduce phospholipid hydroperoxides in vitro by using glutathione as a reductant. This enzyme is widely expressed and is enriched in lungs. A fusion protein of green fluorescent protein with 1-cysPrx was stably expressed in a lung-derived cell line (NCI-H441) lacking endogenous enzyme. Overexpressing cells (C17 or C48) degraded H 2O2 and t-butylhydroperoxide more rapidly and showed decreased sensitivity to oxidant stress as measured by 51

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“…Cell injury was determined by the release of 51 Cr (14). To label cells, 51 Cr (2 ϫ 10 5 cpm per well) was added 24 h before the experiment.…”

Section: Methodsmentioning

confidence: 99%

1-Cys peroxiredoxin overexpression protects cells against phospholipid peroxidation-mediated membrane damage

Manevich

Sweitzer

Pak

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

196

169

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“…This could limit the utility of such conjugates as therapeutic reagents in vivo. However, glucose oxidase conjugates that are active in the presence of catalase have been reported (26). The insensitivity of this particular conjugate to catalase neutralization might be due to the generation of H2O2 intracellularly or in environments that are inaccessible to serum enzymes (19,27).…”

Section: Speciesmentioning

confidence: 99%

Generation of cytotoxic agents by targeted enzymes

et al. 1993

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“…For example, GOX conjugated with collagen antibody caused skin lesions after intradermal injection in rats (24). GOX conjugated with antibodies to endothelial antigens caused oxidative injury in endothelial cell culture (22,23). 125 I-GOX conjugated with monoclonal antibody against angiotensin-converting enzyme (ACE) displayed preferential accumulation in the rat lungs after intravenous injection (29).…”

mentioning

confidence: 99%

Immunotargeting of glucose oxidase to endothelium in vivo causes oxidative vascular injury in the lungs

Christofidou‐Solomidou

Pietra²,

Solomides

et al. 2000

American Journal of Physiology-Lung Cellular and Molecular Phys

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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.

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Specific killing of human endothelial cells by antibody-conjugated glucose oxidase

Cited by 27 publications

References 12 publications

1-Cys peroxiredoxin overexpression protects cells against phospholipid peroxidation-mediated membrane damage

1-Cys peroxiredoxin overexpression protects cells against phospholipid peroxidation-mediated membrane damage

Generation of cytotoxic agents by targeted enzymes

Immunotargeting of glucose oxidase to endothelium in vivo causes oxidative vascular injury in the lungs

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