Macrophage-Derived LIF and IL1B Regulate Alpha(1,2)Fucosyltransferase 2 (Fut2) Expression in Mouse Uterine Epithelial Cells During Early Pregnancy1

Abstract

Macrophages accumulate within stromal tissue subjacent to the luminal epithelium in the mouse uterus during early pregnancy after seminal fluid exposure at coitus. To investigate their role in regulating epithelial cell expression of fucosylated structures required for embryo attachment and implantation, fucosyltransferase enzymes Fut1, Fut2 (Enzyme Commission number [EC] 2.4.1.69), and Fut4 (EC 2.4.1.214) and Muc1 and Muc4 mRNAs were quantified by quantitative real-time PCR in uterine epithelial cells after laser capture microdissection in situ or after epithelial cell coculture with macrophages or macrophage-secreted factors. When uterine macrophage recruitment was impaired by mating with seminal plasma-deficient males, epithelial cell Fut2 expression on Day 3.5 postcoitus (pc) was reduced compared to intact-mated controls. Epithelial cell Fut2 was upregulated in vitro by coculture with macrophages or macrophage-conditioned medium (MCM). Macrophage-derived cytokines LIF, IL1B, and IL12 replicated the effect of MCM on Fut2 mRNA expression, and MCM-stimulated expression was inhibited by anti-LIF and anti-IL1B neutralizing antibodies. The effects of acute macrophage depletion on fucosylated structures detected with lectins Ulex europaeus 1 (UEA-1) and Lotus tetragonolobus purpureas (LTP), or LewisX immunoreactivity, were quantified in vivo in Cd11b-dtr transgenic mice. Depletion of macrophages caused a 30% reduction in luminal epithelial UEA-1 staining and a 67% reduction in LewisX staining in uterine tissues of mice hormonally treated to mimic early pregnancy. Together, these data demonstrate that uterine epithelial Fut2 mRNA expression and terminal fucosylation of embryo attachment ligands is regulated in preparation for implantation by factors including LIF and IL1B secreted from macrophages recruited during the inflammatory response to insemination.

Seminal Fluid Signalling in the Female Reproductive Tract: Implications for Reproductive Success and Offspring Health

“…In different species, effects include clearing the uterine cavity of microorganisms introduced at mating , induction of ovulation and promoting corpus luteum formation (Gangnuss et al 2004 ), supporting development of the pre-implantation embryo , inducing endometrial expression of embryo attachment molecules and angiogenic factors to promote uterine receptivity (Jasper et al 2011 ), and priming the female immune response to paternal antigens (Robertson and Sharkey 2001 ;Moldenhauer et al 2009 ) to promote T cell-mediated immune tolerance (Moldenhauer et al 2009 ) (Fig. 6.1 ).…”

Macrophages regulate corpus luteum development during embryo implantation in mice

“…In the uterus, macrophages fluctuate in number and phenotype over the ovarian cycle, and additional macrophages are recruited into the endometrium in an inflammationlike response to seminal fluid during the pre-implantation period of early pregnancy (8,9). Uterine macrophages are implicated in remodeling processes and in inducing expression of epithelial glycoproteins associated with embryo attachment (10), and they potentially contribute to subsequent events of the uterine decidual response and placental trophoblast invasion (11). Within the ovary, macrophages accumulate within the thecal layer of developing follicles, and there is evidence for a supporting role in follicle growth and ovulation (12)(13)(14) as well as for a contribution to vascular homeostasis (15).…”

TGF-β Mediates Proinflammatory Seminal Fluid Signaling in Human Cervical Epithelial Cells

“…In animals, seminal fluid interacts with epithelial cells lining the female reproductive tract to activate cytokine gene expression and elicit changes in the abundance and behavior of infiltrating leukocyte populations, which, in turn, promote receptivity for embryo implantation (5). In mice, uterine dendritic cells recruited in response to seminal fluid are instrumental in activating local T cell responses (6,7), including expansion of regulatory T (Treg) cell populations that mediate tolerance of the conceptus in an ensuing pregnancy (8), whereas recruited macrophages influence epithelial cell expression of glycosylated structures required for embryo attachment and invasion (9).…”