Soluble Factors From LPS- and PHA-Activated PBMC Induce MAPK, Stat1 and Stat3 Phosphorylation in Primary Cultures of Human Term Placental Trophoblasts: Implications for Infection and Prematurity☆

Abstract 

Infection of the maternal vaginal tract is one of the single most important antecedents of premature labor. We have hypothesized that the abundant local synthesis of pro-inflammatory cytokines that occurs during infection may disrupt the delicate “immunological cross-talk” that must occur between maternal and fetal tissues in order to carry pregnancy to term. These experiments were undertaken as part of a larger study directed at testing that hypothesis.

We prepared primary cultures of human trophoblasts from term placentas. Cell cultures were stimulated with conditioned medium from resting, PHA or LPS-activated peripheral blood mononuclear cells (PBMC). Medium with LPS or PHA at the same concentration as that used to stimulate the PBMC was used as an additional control. Lysates were subjected to western blotting for activated forms of the mitogen-activated protein kinases (MAPK), Stat1, and Stat3.

Western blotting showed phosphorylation of the Jun kinase (JNK), p38, and Erk1/Erk2 MAPK in trophoblasts incubated with conditioned medium from LPS or PHA-activated PBMC but not from medium from resting PBMC, or with PHA or LPS alone. Phosphorylation could be detected as early as 5min and was still observable by 10min, the latest time point tested. Similarly, Stat1 and Stat3 phosphorylation was observed within 10min of exposure to conditioned medium and was still observable 10min after exposure. Immunohistochemistry also demonstrated nuclear translocation of both Stat1 and Stat3 after stimulation of trophoblasts with medium from activated PBMC.

These findings are compatible with the hypothesis that immunologic balance at the maternal-fetal interface is maintained by ongoing “cross-talk” between the fetus (and fetally-derived tissues) and the maternal immune system. Infection of the maternal vaginal tract may disrupt this delicate immunologic balance, initiating inflammatory events that ultimately result in preterm labor.

Keywords: Trophoblasts, Signaling transduction, Inflammation

Abbreviations: FBS, fetal bovine serum, LPS, lipopolysaccharide, PBMC, peripheral blood mononuclear cells, MAPK, mitogen-activated protein kinase, JNK, Jun kinase