Svetlana Vrzic-Petronijevic

Galectin-1 binds mucin in human trophoblast.

Mucins are multifunctional highly glycosylated proteins expressed by the female reproductive tract. Differential expression of MUC1 and MUC15 has been shown in trophoblast. This study was undertaken to establish the distribution of mucin(s) in cytotrophoblast cell cultures using anti-bovine submaxillary mucin (BSM) and to investigate the possibility of MUC1/mucin(s) being a binding partner of trophoblast galectin-1. MUC1 is demonstrated here using immunocytochemistry on isolated cytotrophoblast and the HTR-8/SVneo extravillous trophoblast cell line but detection of additional trophoblast mucins cannot be excluded. Western blot analysis showed similar bands ranging from 30 to >200xa0kDa with anti-BSM and the well-known mucin antibodies HMFG1 and B72.3. Immunocytochemistry and cell-based ELISA data were found to support that all of the antibodies used are reactive with BSM, suggesting the presence of shared epitopes between BSM and trophoblast mucin(s). Binding of galectin-1 to trophoblast MUC1/mucin(s) was analyzed using a solid-phase assay and co-immunoprecipitation. Recombinant galectin-1 binding to isolated trophoblast mucin in solid-phase assay was sensitive to lactose, a carbohydrate inhibitor of galectin binding. In whole HTR-8/SVneo lysates, ~200xa0kDa mucin was detected in galectin-1 immunoprecipitates, while endogenous galectin-1 was present in BSM-immunoprecipitates. Furthermore, double fluorescence immunocytochemistry showed overlap of galectin-1 and trophoblast mucins at the plasma membrane of HTR-8/SVneo cells. These results suggest that trophoblast mucin(s) could act as binding partners of galectin-1, in a carbohydrate-dependent manner.

Immunoglobulins from sera of APS patients bind HTR-8/SVneo trophoblast cell line and reduce additional mediators of cell invasion

Immunoglobulins from sera of patients with antiphospholipid syndrome (APS) decrease trophoblast cell invasion in vitro. This study aimed to extend understanding of cellular effects of immunoglobulins from APS (aPL+) in HTR-8/SVneo cells. aPL+ IgG induced change in effector molecules important for cell invasion was investigated further. After 1h of culture 21% cells bound aPL+ IgG, as opposed to 6% in control (aPL-). This was accompanied by increase in phospho-p38 at 30min. After 24h treatment aPL+IgG decreased protein levels of integrin subunits α1 (78% of control; p<0.01), α4 (65% of control, p<0.01), α5 (76% of control; p<0.01) and β1 (80% of control; p<0.01), and secreted gal-1 (68% of control; p<0.05). ProMMP-9 was reduced to 70% of control (p<0.001). Treatment with inhibitor of p38 MAPK signaling SB202190 reversed inhibition in integrin β1 and secreted gal-1. Involvement of p38 MAPK signaling and decrease in integrin subunit α4, proMMP-9, and secreted gal-1 in HTR-8/SVneo cells are novel and extend the list of mediators of trophoblast invasion affected by aPL.

Immunoglobulins from sera of antiphospholipid syndrome patients are internalized in the HTR-8/SVneo cell line and cytotrophoblast in culture

Women with antiphospholipid syndrome (APS) experience pregnancy complications mostly due to impaired trophoblast cell functions. Antiphospholipid antibodies (aPL) affect extravillous trophoblast in vivo and in culture, but the mechanisms are still poorly understood. Previously, syncytiotrophoblast was shown to bind and internalize aPL, which was not replicated for extravillous cytotrophoblast in short term culture. Here, aPL binding and time dependent internalization was demonstrated with exposure to aPL in the extravillous cell line HTR-8/SVneo and isolated first trimester of pregnancy cytotrophoblast (CT) using immunocytochemistry and flow cytometry. Intracellular aPL were detectable from 2u202fh of culture, reaching 30.7u202f±u202f3.1% (pu202f<u202f0.001) positive cells in CT and 24.8u202f±u202f7% (pu202f<u202f0.01) in HTR-8/SVneo cells at 24u202fh and 33u202f±u202f4.2% (pu202f<u202f0.01) at 48u202fh. The data presented show that extravillous trophoblast cells internalize aPL in a time-dependent manner significantly more than control immunoglobulins after 24u202fh of exposure.

First trimester ultrasonographic parameters in prediction of the course and outcome of monochorionic twin pregnancies

BACKGROUND/AIMnThe aim of the study was to evaluate the association and the potential predictive value of first trimester ultrasonographic parameters on the course and outcome of monochorionic diamniotic twin pregnancies.nnnMATERIALS AND METHODSnA prospective cohort study was undertaken of 39 healthy women with consecutive monochorionic diamniotic twin pregnancies. During first-trimester screening, crown-rump length (CRL) and nuchal translucency (NT) were measured. The intertwin discordance in CRL and NT was determined. As pregnancy outcomes we assessed twins live-born rates, Apgar scores, birth weight, pregnancy complications, and gestational week of delivery.nnnRESULTSnNone of the assessed pregnancy outcomes significantly correlated with standard CRL discordance ≥10%. The newly established cut-off was 3.75 mm for CRL and 1.3 mm for NT. Monochorionic diamniotic twins were delivered in a later gestational week and had better chance of survival if CRL intertwin difference was <3.75 mm. Apgar scores significantly negatively correlated only with NT of corresponding twins. When intertwin NT difference was ≥1.3 mm, twins had lower birth weight and pregnancy complications were more frequent. Regression models show that intertwin CRL difference <3.75 mm is a significant predictor of live-born monochorionic diamniotic twins.nnnCONCLUSIONnCRL and NT in monochorionic diamniotic twin pregnancies could indicate pregnancy complications and outcomes.