Role of intrauterine administration of transfected peripheral blood mononuclear cells by GM-CSF on embryo implantation and pregnancy rate in mice.

Abstract

One of the effective treatments in women with recurrent implantation failure (RIF) is the use of immune cells to facilitate embryo implantation. Previous studies have shown that intrauterine transmission of peripheral blood mononuclear cells (PBMC) increased the embryo implantation rate. In this study using B6D2F1 (C57BL/6 × DBA2) mice, a fragment of the granulocyte macrophage colony-stimulating factor (Gm-csf) gene was cloned into an enhanced green fluorescent protein vector (pEGFP-N1) and then transfected into PBMC. The protein level of GM-CSF was evaluated in the transfected PBMC and untransfected PBMC by ELISA. Attachment of mouse embryos and the mRNA expression levels of leukemia inhibitory factor (Lif), vascular endothelial growth factor (Vegf), matrix metalloproteinase 9 (Mmp9), Gmcsf-receptor (Gmcsf-r) and interleukin 6 (Il6) in vitro were assessed by real-time PCR in endometrial cells. To determine the pregnancy rate and number of implantation sites in vivo, the mouse uterine horns were analyzed on Day 7.5 post coitum. A greater amount of GM-CSF was produced in PBMC transfected with recombinant vector (552pg/mL) compared with the untransfected PBMC (57pg/mL) and PBMC transfected with empty vector (34pg/mL) (P< 0.05). The data showed that the embryo attachment rate and mRNA expression levels (Vegf [1.7-fold], Mmp9 [1.4-fold], Lif [1.5-fold], Gm-csf r [1.6-fold] and Il6 [1.2-fold]) in the in vitro study (P< 0.01), pregnancy rate (P< 0.01) and number of implantation sites (P< 0.01) in the in vivo investigation (P< 0.05) were increased in PBMC transfected with recombinant vector compared with the PBMC group. The study demonstrated that, in mice, endometrium immunotherapy with transfected PBMC that contained recombinant GM-CSF before embryo implantation was effective in improving embryo implantation and endometrial receptivity.