Blocking both E-selectin and P-selectin inhibits neutrophil recruitment into the murine testis after ischemia-reperfusion-induced 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 mu g function-blocking monoclonal E-selectin and P-selectin antibody (FBMAb group; 100 mu g; each) intravenously or 200 mu g of isotype-matched control-antibody (IMCAb group). Separate groups of mice underwent sham-operation (SO group) or received 500 ng of TNF alpha (IF group) to induce inflammation. Mice were sacrificed 24 It 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.