Equine seminal plasma reduces sperm binding to polymorphonuclear neutrophils (PMNs) and improves the fertility of fresh semen inseminated into inflamed uteri

Abstract

Seminal plasma (SP) is known to have immunosuppressive properties in several species. Equine SP has been reported to reduce or inhibit chemotaxis, phagocytosis and complement activityin vitro. The type and amount of the SP component that suppresses sperm–polymorphonuclear neutrophil (PMN) bindingin vitrowas determined, and the effect of such suppression on the fertility of mares inseminated in the presence of uterine inflammation, was analyzed. Sperm cells were suspended in either SP, semen extender or a mixture of both, and each was mixed with PMN-rich uterine secretions collected at 12 h after artificial insemination (AI). SP reduced binding between spermatozoa and PMNs significantly (P< 0.05). Fertile spermatozoa were suspended in SP or semen extender and used to inseminate mares 12 h after the induction of uterine inflammation. The pregnancy rate was normal (77%) when spermatozoa were suspended in SP, but was dramatically reduced to only 5% when spermatozoa were suspended in extender. The proteins from SP, blood plasma (BP) and a skim-milk-based semen extender (skim milk extender, SME) were precipitated by ammonium sulfate, resuspended in PBS and dialyzed. The effect of the precipitated proteins on sperm–PMN binding was compared with fresh, untreated SP. Both fresh SP, and isolated SP proteins reduced sperm–PMN binding (P< 0.001). Conversely, proteins isolated from either BP or SME did not reduce sperm–PMN binding. The different concentrations of SP proteins used showed a dose-dependent suppression of sperm–PMN binding. Concentrations of 1 mg/ml SP protein significantly reduced sperm–PMN binding and 6 mg/ml reduced the binding to a level similar to that observed with fresh whole SP (P< 0.001). Finally, SP protein digested with proteinase K resulted in the complete loss of SP suppressive activity confirming that the effective component is a proteinaceous substance.