Abstract
High rate of fetal mortality in ruminant somatic cell nuclear transfer (SCNT) pregnancies is due, at least in part, to immune-mediated abortion of fetuses. In the present study, goat uterine leukocytes were isolated by Dolichos biflorus agglutinin (DBA) coated magnetic beads, and with majority being were CD56+CD16- in phenotype with low levels of perforin and Granzyme B expression. The responses of the isolated cells to SCNT and in vitro fertilization (IVF) embryos conditioned mediums containing hormone steroids were compared by measuring their phenotype and cytokines expression. The results showed there was a 2-fold increase in the numbers of isolated uterine leukocytes after incubation with different conditioned mediums for 120 h. However, significantly lower percentage and absolute numbers of uterine CD56+CD16- leukocytes incubated with SCNT conditioned mediums were detected as compared with those incubated with IVF conditioned mediums (P < 0.05). The group treated with progesterone (P4) or the combination of P4 and 17β-estradiol (E2) were associated with significantly higher percentage and absolute numbers of CD56+CD16- cells as compared with those treated with E2 alone (P < 0.05). Furthermore, in the presence of steroids, the isolated leukocytes incubated with SCNT conditioned mediums associated with greater levels of IFN-γ secretion and expression, as well as lesser levels of VEGF, as compared with those treated with IVF conditioned mediums (P < 0.05). In conclusion, this study demonstrates that SCNT embryos have a profound effect on the phenotype expression of goat uterine DBA+ leukocytes, as well as the secretion and expression of IFN-γ and VEGF by these cells in vitro.
Highlights
The ability to produce clones by somatic cell nuclear transfer (SCNT) has improved significantly in the last decade
in vitro fertilization (IVF) embryos were used as a control for the identification of specific responses of the uterine Dolichos biflorus agglutinin (DBA)+ leukocytes derived from non-pregnant endometrium to SCNT embryos
DBA lectin reactivity has been widely used for histological recognition and isolation of uterine NK cells in pregnant mice [14, 15]
Summary
The ability to produce clones by somatic cell nuclear transfer (SCNT) has improved significantly in the last decade. The first trimester losses of more than 50% of the transferred embryos are common for SCNT pregnancies in cattle, sheep, and goats [1, 2]. Uterine Leukocytes Respond to Cloned versus Fertilized Embryos the majority of fetal rejection in SCNT pregnancies in ruminants is due to immune-mediated placental rejection [3, 4]. Uterine natural killer (uNK) cells represent a distinct lymphocyte subset with a central role in immune regulation during pregnancy. An inadequate uterine environment induced by insufficient steroid concentration during early pregnancy in ewes can cause early embryonic death [9]
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