Anti-CD11d antibody treatment reduces free radical formation and cell death in the injured spinal cord of rats.
Treatment with a monoclonal antibody (mAb) against the CD11d subunit of the leukocyte integrin CD11d/CD18 after spinal cord injury (SCI) decreases intraspinal inflammation and oxidative damage, improving neurological function in rats. In this study we tested whether the anti-CD11d mAb treatment reduces intraspinal free radical formation and cell death after SCI. Using clip-compression SCI in rats, reactive oxygen species (ROS) generated in injured spinal cord were detected using 2',7'-dichlorofluorescin-diacetate and hydroethidine as fluorescent probes. ROS in the injured cord increased significantly after SCI; anti-CD11d mAb treatment significantly reduced this ROS formation. Immunohistochemistry and western blotting were employed to assess the effects of anti-CD11d mAb treatment on spinal cord expression of gp91Phox (a subunit of NADPH oxidase producing superoxide) on formation of 4-hydroxynonenal (HNE, indicating lipid peroxidation) and on expression of caspase-3. We also assessed effects on cell death, determined by cell morphology. The expression of gp91Phox, formation of HNE, and cell death increased after SCI. Anti-CD11d mAb treatment clearly attenuated these responses. In conclusion, anti-CD11d mAb treatment significantly reduces intraspinal free radical formation caused by infiltrating leukocytes after SCI, thereby reducing secondary cell death. These effects likely underlie tissue preservation and improved neurological function that result from the mAb treatment.
Spinal Cord Injury Team, BioTherapeutics Research Group, Robarts Research Institute, London, Ontario, Canada. fbao@robarts.ca
Feng
F
Bao
Gregory A
GA
Dekaban
Lynne C
LC
Weaver
D000447:Aldehydes / Q000037:antagonists & inhibitors; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D018845:CD11 Antigens / Q000276:immunology*; D016923:Cell Death / Q000187:drug effects; D004996:Ethidium / Q000037:antagonists & inhibitors; D005260:Female; D005452:Fluoresceins / Q000493:pharmacokinetics; D005609:Free Radicals / Q000037:antagonists & inhibitors*; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors; D051381:Rats; D017208:Rats, Wistar; D013117:Spinal Cord Compression / Q000150:complications; D013119:Spinal Cord Injuries / Q000209:etiology / Q000378:metabolism / Q000503:physiopathology*; D013481:Superoxides / Q000037:antagonists & inhibitors
D000447:Aldehydes; D000911:Antibodies, Monoclonal; D018845:CD11 Antigens; D005452:Fluoresceins; D005609:Free Radicals; C503911:Itgad protein, rat; D008562:Membrane Glycoproteins; D013481:Superoxides; C037631:2',7'-dichlorofluorescein; C500593:Cybb protein, rat; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; D004996:Ethidium; C027576:4-hydroxy-2-nonenal