Acta Vet. Brno 2008, 77: 321-326

Blocking Both E-Selectin and P-Selectin Inhibits Neutrophil Recruitment into the Murine Testis after Ischemia-Reperfusion-Induced Injury

M. Celebi1,2, A. G. A. Paul3

1Cardiovascular Division, University of Virginia School of Medicine, Charlottesville, VA, USA
2Department of Reproduction, Faculty of Veterinary Medicine, University of Ondokuz Mayis, Samsun, Turkey
3Department of Pathology, University of Virginia School of Medicine, Charlottesville, VA, USA

Received September 28, 2007
Accepted June 11, 2008

Ischemia-reperfusion (IR) injury of the testis results in germ cell specific apoptosis, a process in which neutrophil recruitment to the testes plays a critical role. Adhesion molecules, in particular E- and P-selectins, play a critical role in this recruitment. The present study sought to characterize the inhibitory effect of function-blocking monoclonal anti-mouse E- and P-selectin antibodies on the migration of neutrophils into the IR-induced testis of the mouse. Mice were subjected to a 2 hr period of testicular torsion (ischemia) followed by detorsion (reperfusion). Ten minutes after the onset of reperfusion mice received either a mixture of 200 μg function-blocking monoclonal E-selectin and P-selectin antibody (FBMAb group; 100 μg; each) intravenously or 200 μg of isotype-matched control-antibody (IMCAb group). Separate groups of mice underwent shamoperation (SO group) or received 500 ng of TNFα (IF group) to induce inflammation. Mice were sacrificed 24 h after reperfusion and testicular interstitial cells were isolated and analyzed for the presence of neutrophils by means of flow cytometry. The mixture of function-blocking monoclonal E- and P-selectin antibody (FBMAb) decreased neutrophil recruitment to the IR-induced testis significantly (FBMAb group as compared to the IMCAb group 20.2 ± 2.8 vs. 51.9 ± 4.0 % Gr-1+CD11b+ of total leukocytes; p = 0.0002). Therefore, blocking both E- and P-selectin may be therapeutically beneficial to protect postischemic testis.