Milos Petronijevic

Effects of anti-phospholipid antibodies on a human trophoblast cell line (HTR-8/SVneo).

Antibodies to phospholipids (aPL) have been shown to adversely affect trophoblast invasion in vivo and in vitro. HTR-8/SVneo cells derived from first trimester of pregnancy extravillous trophoblast were studied. Matrigel invasion assay, cytochemistry and cell-based enzyme-linked immunosorbant assay (ELISA) with aPL or normal IgG was used. Our data show that aPL at 100 microg/ml decrease invasiveness of HTR-8/SVneo cells to 60% of control (p<0.01), and this was also shown for primary cytotrophoblast (to 15.5% of control, p<0.001). aPL treatment caused a significant decrease in integrin alpha(1), alpha(5), and beta(1) proteins (86%, 84%, and 87%, respectively). We conclude that HTR-8/SVneo cell culture is a suitable model to study mechanisms of action of aPL on trophoblast, which in HTR-8/SVneo cells inhibit invasion by decreasing integrins alpha(5), alpha(1), and beta(1).

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.

Choriocarcinoma cell line Response to Dexamethasone

Choriocarcinoma cell line Response to Dexamethasone Choriocarcinoma cell lines JAr and JEG-3 are model systems for the study of transformed trophoblast. Both cell lines were shown to produce galectin-1, expression of which was increased in choriocarcinoma when compared to the normal trophoblast of pregnancy. In this study the effects of synthetic glucocorticoid dexametha-sone were investigated in both JAr and JEG-3 cell lines by the MTT test, cell based ELISA, and the cell adhesion and migration tests. Viable cell number/cell proliferation of JAr cells was significantly increased after treatment with 0.1 and 1 nmol/L of dexamethasone, while proliferation of JEG-3 cells was significantly increased after treatment in the whole concentration range of dexamethasone (0.1-100 nmol/L). Galectin-1 in JAr cells was modulated by dexamethasone, which mildly, but significantly decreased production at low concentrations (0.1 and 1 nmol/L). In JEG-3 cells production of galectin-1 was significantly decreased only after treatment with 100 nmol/L of dexamethasone. Cell adhesion of JEG-3 was significantly increased in the presence of lactose, an inhibitory sugar for gal-1, while dexamethasone induced decrease of JEG-3 cell migration. These findings have shown that dexamet-hasone may affect proliferation, gal-1 production and cell migration, in a cell line specific manner. These data suggest that glucocorticoid treatment in vivo might have the potential to affect cell functions in choriocarcinoma. Odgovor Horiokarcinomskih Ćelijskih Linija na Tretman Deksametazonom Horiokarcinomske ćelijske linije JAr i JEG-3 su model sistemi za ispitivanje transformisanog trofoblasta. Obe ćelijske linije eksprimiraju galektin-1, čija je povećana ekspresija pokazana u horiokarcinomima u odnosu na normalni trofoblast trudnoće. U ovom radu ispitivan je efekat sintetskog glukokortikoida deksameta-zona na JAr i JEG-3 ćelijske linije, upotrebom MTT testa, ELISA testa na ćelijama i testovima adhezije i migracije. Broj živih ćelija, kao indikator proliferacije ćelija, značajno je povećan nakon tretmana 0,1 i 1 nmol/L deksametazonom, a proliferacija JEG-3 je značajno povećana u čitavom opsegu upotrebljenih koncentracija deksametazona (0,1-100 nmol/L). Galektin-1 je u JAr moduliran deksa-metazonom, koji blago, ali značajno smanjuje produkciju galektina-1 u niskim koncentracijama (0,1 i 1 nmol/L). Kod JEG-3 ćelija, produkcija galektina-1 je značajno smanjena samo nakon tretmana 100 nmol/L deksametazonom. U prisustvu laktoze kao inhibitornog šećera za ga-lektin-1 adhezija JEG-3ćelija je značajno povećana, dok deksametazon uz to smanjuje i migraciju JEG-3 ćelija. Rezultati dobijeni u ovom radu pokazuju da deksametazon utiče na proliferaciju i u manjoj meri na galektin-1 kod JAr i JEG-3 ćelija, na način specifičan za ćelijsku liniju. Dobijeni podaci ukazuju na to da bi tretman glukokortikoidima in vivo mogao imati uticaja na ćelije horiokarcinoma.

P22.01: Fetal echocardiography and pregnancy outcome in Serbia

Results: Low VCI at the entry was found in 19 patients (0.3% of 6024 screenings). Uterine isthmus had already been opened at the entry in 12 patients. Nine cases of VP and one of low VCI without VP were diagnosed at the entry and consistent until the delivery. Two cases were diagnosed as placenta previa with low VCI at the entry. They progressed to low-lying placenta with VP at delivery due to atrophy of the placenta around the internal os. On the other hand, seven patients were diagnosed as low VCI near the closed isthmus at the entry. Five were diagnosed as low VCI without VP at the delivery, however, two cases of velamentous vessels were descending toward the isthmus with the expansion of the amniotic bag after the opening of the isthmus, resulting in VP at the delivery. Conclusions: Not only atrophy of the placenta previa around os develops VP, but also low VCI occasionally migrates in the direction of the uterine cervix with uterine isthmus opening, resulting in VP.