Effect of oestrous cycle and early pregnancy on uterine production and expression of immune regulatory factors in gilts

Abstract

This study was undertaken to characterize uterine immune factors involved in the establishment of pregnancy in gilts. Thirty crossbred Yorkshire–Landrace gilts of similar age and weight were observed twice a day for oestrous behaviour with intact boars. On the day of first standing oestrus (Day 0) and 12 h later, 15 gilts were inseminated with pooled semen from Duroc boars of proven fertility. Pregnant gilts were slaughtered either on Days 10, 15 or 25 of gestation ( n=5 per day). The other 15 gilts were not inseminated and were slaughtered on either Days 0, 10 or 15 of the oestrous cycle ( n=5 per day). Immediately after slaughter, endometrial tissue samples from the mesometrial side were removed for gene expression using RNase protection assay and in situ hybridization methodologies. The other uterine horn was flushed with 20 ml of PBS to collect the uterine fluid. In pregnant gilts, endometrial interleukin (IL)-6 mRNA expression was higher on Day 15 than on Days 10 and 25 ( P<0.01 and P<0.1, respectively). On Day 15, IL-6 expression was also significantly higher ( P<0.01) in pregnant gilts than in cyclic gilts. In both pregnant and cyclic gilts, transforming growth factor (TGF)-β2 in uterine fluid was significantly higher ( P<0.0001) on Day 15 than on Day 10. At the gene expression level, TGF-β2 also increased between Days 10 and 15 in both cyclic and pregnant gilts but differences were not significant. On Day 15, concentrations of interferon-γ and prostaglandin E 2 (PGE 2) in uterine fluid were markedly higher ( P<0.001) in pregnant gilts than in cyclic gilts, whereas the total amount of TGF-β2 in uterine fluid and its endometrial expression were approximately 70% higher although this increase was not significant. Finally, tumour-necrosis factor-α and granulocyte-macrophage/colony-stimulating factor mRNA expressions were undetectable in all endometrial samples. In conclusion, production and/or expression of uterine TGF-β2, IL-6 and PGE 2 increased during the embryonic attachment period and are coincidental with embryonic interferon-γ production.