Blocking Both E-Selectin and P-Selectin Inhibits Neutrophil Recruitment Into the Murine Testis After Ischemia-Reperfusion Injury

Abstract

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 sham-operation (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.