Porcine spermatozoa inhibit post-breeding cytokine induction in uterine epithelial cells in vivo.
Early endometrial cytokine responses after exposure to various inseminate components were investigated for a better understanding of the immunological reactions occurring in the porcine uterus after insemination. Baseline values were established for the mRNA concentrations of GM-CSF, IL-6, IL-10, CXCL8 (interleukin-8), Tumour Necrosis Factor alpha (TNF-alpha), TGF-beta, cyclooxygenase-2 (COX-2) and arachidonate 5-lipooxygenase (ALOX-5) in periovulatory uterine endometrial tissue using quantitative RT-PCR. Synchronized gilts were inseminated with spermatozoa diluted either in the semen extender Androhep or seminal plasma. Uterine infusions of media without spermatozoa were used as controls. Three hours after insemination sows were slaughtered and the expression of the above mentioned cytokines was measured in uterine epithelial cells. Simultaneously, the influx of polymorphonuclear neutrophilic (PMN) granulocytes into the uterus was quantified. Compared to baseline values seminal plasma (SP) and Androhep (AH) respectively, if used alone, caused a significant increase in mRNA concentrations of IL-10 (SP: 1.5-fold), TGF-beta (AH: 1.5-fold), CXCL8 (AH: 7.1-fold), TNF-alpha (AH: 1.9-fold) and COX-2 (AH: 7-fold). Surprisingly, in the presence of spermatozoa, none of the tested cytokines revealed mRNA concentrations higher than baseline values. The number of immigrated, intra-luminal PMN correlated only with mRNA concentrations of CXCL8 in presence of Androhep (r=0.51). None of the other cytokines tested seemed to be involved in the regulation of neutrophil recruitment. However, the most interesting result was the sperm-induced down-regulation in the expression of TNF-alpha, TGF-beta, IL-10, CXCL8 and COX-2 to mRNA concentration levels similar to or even below baseline values. In conclusion the results show that CXCL8 contributes significantly to uterine PMN recruitment and indicate a so far underestimated role of porcine spermatozoa in the general regulation of the uterine post-mating inflammatory response.
Taylor
Ulrike
U
Institute of Farm Animal Genetics, Friedrich Loeffler Institute, Federal Research Institute for Animal Health, 31535 Neustadt, Germany.
Zerbe
Holm
H
Seyfert
Hans-Martin
HM
Rath
Detlef
D
Baulain
Ulrich
U
Langner
Kathrin Friederike Annika
KF
Schuberth
Hans-Joachim
HJ
D000818:Animals; D001094:Arachidonate 5-Lipoxygenase / Q000235:genetics; D001947:Breeding; D051546:Cyclooxygenase 2 / Q000235:genetics; D016207:Cytokines / Q000096:biosynthesis*; D004717:Endometrium / Q000502:physiology; D004847:Epithelial Cells / Q000502:physiology*; D005260:Female; D006098:Granulocytes / Q000502:physiology; D007315:Insemination, Artificial / Q000662:veterinary; D007378:Interleukins / Q000235:genetics; D008297:Male; D009504:Neutrophils / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics; D012661:Semen / Q000502:physiology; D013094:Spermatozoa / Q000502:physiology*; D013552:Swine; D016212:Transforming Growth Factor beta / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics; D014599:Uterus / Q000502:physiology*
D016207:Cytokines; D007378:Interleukins; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D001094:Arachidonate 5-Lipoxygenase; D051546:Cyclooxygenase 2