Changes in mononuclear immune cells during bovine pregnancy.

This study aimed to determine the differences in gene expression between mononuclear cells derived from peripheral blood and endometrium during pregnancy in cattle and to determine the proportion of mononuclear cells in the endometrium of pregnant and diestrous cows. Endometrial tissue and peripheral blood were collected from Day 34±2 pregnant cows, and mononuclear cell populations were quantified and sorted (n =5). The relative mRNA levels of inflammatory mediators was assessed by quantitative real time polymerase chain reaction. During pregnancy, the proportion of CD8+ , CD4+ , CD4+ CD25- and CD4+ CD25dim cells among mononuclear cells was greater in blood than endometrium, and cells positive for CD14 and CD68 expressed greater mRNA amounts of interleukin (IL ) 6 , CXCL8 and IL10 in endometrium compared with blood. Cells positive for γ/δ-T cell receptor expressed greater amounts of IL1A transcript in the endometrium than in blood of diestrous cows, CD4+ CD25bright cells expressed more CTLA4 mRNA in the endometrium compared with blood of diestrous cows, and endometrial natural killer cells expressed greater CXCL8 mRNA compared with blood of pregnant and diestrous cows. The percentages of CD21+ , NCR1+ , CD8+ , FoxP3+ , CD3+ and CD68+ cells were greater in the endometrium of Day 35 pregnant cows compared with diestrous cows.

Lymphocyte soluble factors from pregnant cows modulate mRNA transcript abundances encoding for proteins associated with trophoblast growth and development.

This study was conducted to determine whether T cell populations are responsible for modulating placental development during gestation in cattle. It was hypothesized that CD4+CD25+ and γ/δ+ T cells modulate gene expression, based on mRNA transcript abundances, and promote proliferation and survival of trophoblast cells. Peripheral blood was collected from cows at 160 to 180 days of gestation and non-pregnant cows, T cell populations CD8+, CD4+, CD4+CD25+, CD24+CD25-, and γ/δ+ T cells were isolated, cultured for 48 h, and supernatant was collected. Placental samples were digested, and trophoblast cells were cultured for 24 h. Trophoblast cells were cultured with 50 µL of T cell-conditioned media and 50 µL of fresh culture media for an additional 48 h. Samples in control wells were treated with unconditioned media. Trophoblast cell proliferation, apoptosis, and mRNA transcript assays were conducted. There was no effect of T cell population on trophoblast apoptosis rate, proliferation, and relative mRNA transcript abundances. The T cell supernatant from pregnant and non-pregnant cows induced greater apoptosis rates in trophoblast cells than unconditioned media. Trophoblast cells proliferated less when treated with T cell supernatant from pregnant compared to unconditioned medium and non-pregnant cows. Treatment with the T cell supernatant from pregnant cows resulted in larger abundances of BMP5, IGF1R, PAG10, FGF2, RSPO3 and TMED2 and also a lesser abundance of FGF2 mRNA transcript than non-pregnant group and unconditioned media treatments. Supernatant from T cell derived from pregnant cows modulates trophoblast mRNA transcript abundances differently from T cell supernatant of non-pregnant cows.