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Updated: Apr 3, 2026
Inducing Ischemia-reperfusion Injury in the Mouse Ear Skin for Intravital Multiphoton Imaging of Immune Responses
Published on: December 22, 2016
Anti-P-selectin monoclonal antibody attenuates reperfusion injury to the rabbit ear
R K Winn1, D Liggitt, N B Vedder
1Department of Surgery, University of Washington School of Medicine, Seattle 98104.
Abstract:
Neutrophil adherence and/or aggregation has been implicated in ischemia reperfusion injuries. We examined the role of P-selectin in PMN-mediated injury after reperfusion of the rabbit ear. The ear was partially amputated, and then reattached leaving the central artery and vein intact. To induce ischemia the central artery was then occluded. Treatment was at reperfusion with either saline or one of two murine P-selectin mAbs, designated PB1.3 and PNB1.6 mAb PB1.3 cross-reacts with rabbit P-selectin and prevents histamine-induced leukocyte rolling, whereas PNB1.6 does not. Using a peroxidase-antiperoxidase system P-selectin was detected in the ischemic ear, but not in the nonischemic ear. Ear volume increased to 5.3 times baseline in the saline-treated animals (n = 8), 6.6 times baseline in the nonblocking mAb PNB1.6-treated animals (n = 2), and 3.7 times baseline in the blocking mAb PB1.3-treated animals (n = 8). Estimated tissue necrosis of the combined saline- and PNB1.6-treated animals was 46 vs. 2.7% for the mAb PB1.3-treated animals. We conclude that: (a) P-selectin is expressed in ischemia reperfusion; (b) P-selectin participates in PMN-endothelial cell interactions in ischemia reperfusion; and (c) inhibiting P-selectin adhesion significantly reduces reperfusion injury.
Insights
P-selectin plays a key role in ischemia reperfusion injury by mediating neutrophil interactions. Blocking P-selectin adhesion significantly reduces tissue damage, highlighting its therapeutic potential.
Area of Science:
- Biomedical Science
- Immunology
- Vascular Biology
Background:
- Neutrophil adherence and aggregation are implicated in ischemia reperfusion injuries.
- P-selectin's role in polymorphonuclear leukocyte (PMN)-mediated injury requires further elucidation.
Purpose of the Study:
- To investigate the role of P-selectin in rabbit ear ischemia reperfusion injury.
- To assess the efficacy of P-selectin blockade in mitigating reperfusion-induced tissue damage.
Main Methods:
- A rabbit ear model was used, involving partial amputation, reattachment, and arterial occlusion to induce ischemia.
- Treatment with saline, a non-blocking P-selectin mAb (PNB1.6), or a blocking P-selectin mAb (PB1.3) was administered at reperfusion.
- P-selectin expression was detected using a peroxidase-anti-peroxidase system, and tissue injury was quantified by ear volume increase and necrosis percentage.
Main Results:
- P-selectin was detected in the ischemic ear but not the non-ischemic ear.
- Blocking P-selectin with mAb PB1.3 significantly reduced ear swelling (3.7x baseline) and tissue necrosis (2.7%) compared to saline (5.3x baseline, 46% necrosis) and non-blocking mAb PNB1.6 (6.6x baseline, 46% necrosis) treatments.
- These findings indicate P-selectin's involvement in PMN-endothelial cell interactions during reperfusion.
Conclusions:
- P-selectin is expressed during ischemia reperfusion.
- P-selectin actively participates in PMN-endothelial cell interactions following reperfusion.
- Inhibition of P-selectin-mediated adhesion offers a significant protective effect against ischemia reperfusion injury.
