Abstract

Sensitization with conceptus antigens has been shown to be useful for improving reproductive performance facilitating maternal acceptance of an allogeneic embryo through the induction of cytokines and immunoregulatory cells in the uterine microenvironment. As FOXP3, IDO, IL10 and CSF1 in the uterus are important on the recognition and development of embryos during early pregnancy, this study aimed to determine whether simultaneous or isolated administration of paternal (semen) and maternal (PBMCs) antigens in the uterus of cow, on the day of estrus, influence the gene expression of these cytokines. Forty crossbred cows were divided into four treatments: T0: Control; T1: Semen; T2: PBMCs (peripheral blood mononuclear cells) from another cow and T3: PBMCs+Semen. Antigens were administered into the uterine body on the estrus day (D0). Uterine biopsies designed for molecular analysis of gene expression were collected in vivo seven (D7) and fourteen (D14) days after immunostimulation. Transcripts from FOXP3, IDO, IL-10 and CSF-1 were detected in all RNA samples extracted from uterine biopsies. The semiquantitative analysis showed that none of the treatments caused significant increase in the expression of these genes. Furthermore, on D14 all treatments led to a decline in the number of CSF-1 transcripts; moreover, treatment with PBMCs+Semen also led to a drop in the abundance of IL-10 transcripts. Such results suggest that isolated or simultaneous administration of both antigens would not increase maternal tolerance to embryo alloantigens, nor would it create favorable conditions to its growth and pre-implantation development, at least regarding the effects mediated by these genes on D7 and D14 of the estrous cycle.

Highlights

  • Improving the bovine embryo transfer technique (ET) is strongly encouraged by the breeding industry need to increase genetic development (MERTON et al, 2003)

  • As FOXP3, IDO, IL10 and CSF1 in the uterus are important on the recognition and development of embryos during early pregnancy, this study aimed to determine whether simultaneous or isolated administration of paternal and maternal (PBMCs) antigens in the uterus of cow, on the day of estrus, influence the gene expression of these cytokines

  • Uterine gene expression of FOXP3, IDO, Interleukin 10 (IL-10) and CSF-1 during early pregnancy seems to be a consequence of immunomodulatory mechanism adjustments during pregnancy that favor the establishment of appropriate intrauterine environment to tolerate the development of an allogeneic embryo

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Summary

Introduction

Improving the bovine embryo transfer technique (ET) is strongly encouraged by the breeding industry need to increase genetic development (MERTON et al, 2003). Most pregnancy losses take place between the 7th and 16th day of gestation (BERG et al, 2010), a period in which the bovine embryo remains free and floating in the uterine lumen, completely dependent on the intrauterine environment to survive and to start its preimplantation growth and susceptible to death from changes in the suitability of this environment during ET (KIMURA; MATSUYAMA, 2014) During this second week, the embryonic mortality after embryo transfer produced in vitro or in vivo is, on average, almost twice as high compared to the one derived from embryos originating from natural mating or artificial insemination (BERG et al, 2010). Pregnancy loss in recipients can be attributed to the interactions between the maternal uterus and the pre-implantation embryo (fully allogeneic), leading to failure in maternal recognition of pregnancy and/or mother intolerance of embryo alloantigens (IDETA et al, 2010a, b; MINTEN et al, 2013).

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