Shiga toxin type 2 impairs trophoblast cell migration but not cell viability.

SCALISE MARÍA LUJÁN; SACERDOTI FLAVIA; IBARRA CRISTINA

Mar del Plata

Congreso; LXI Reunión Anual de la Sociedad Argentina de Investigación Clínica, LXIV Reunión Anual de la Sociedad Argentina de Inmunología y XLVIII Reunión Anual de la Sociedad Argentina de Farmacología Experimental.; 2016

Sociedad Argentina de Investigación Clínica

Shiga toxin type 2 (Stx2) is the main virulence factor of Shigatoxin producing Escherichia coli (STEC). STEC are pathogensinvolved in food-borne diseases and are responsible for Hemo-lytic Uremic Syndrome (HUS) development. We have previouslydemonstrated that Stx2 induce miscarriage and premature deliveryin rats. We propose that STEC infections during early pregnancymay cause damage in the development of placenta mediated byStx2. The aim of this study was to evaluate the effects of Stx2,alone or in combination with lipopolysaccharide (LPS), on humanfirst trimester trophoblast cells. In order to analyze if viability orcell migration could be affected by Stx2, citotoxicity and wound-healing assays were performed. HTR-8 and Swan 71 first trimestertrophoblast cells were exposed to different concentrations of pureStx2 (1 ng to 1 μg/ml), with or without LPS (5 μg/ml). Cell viabilitywas analyzed by neutral red uptake at 72 h after treatment. Onthe other hand, HTR-8 cells were seed in 24 well plates andpre-incubated for 24 h in arrested conditions with the differentconcentrations of Stx2 (1-10 ng/ml) with or without LPS to estimatethe effect of Stx2 on extent of cell migration. After that cells werewashed with PBS and a vertical scratch was made in the centerof each well. Photos of the scratch were taken at regular times (0,5, 24 h). Images were analyzed by TScratch software version 1.0to estimate the percentage of bound closure. Stx2 did not affect HRT-8 or Swan 71 cell viability even in combination with LPS.However Stx2 impaired HTR-8 cell migration after 24 h of treat-ment with Stx2 alone or in combination with LPS. These resultssuggest that Stx2 can alter in vitro cell trofoblast migration. Thesedata suggest that Stx2 may affect trophoblast invasion and earlyplacentation. Although nowadays there are not reports indicatingthat Stx2 may affect early pregnancy in humans this data suggesta possible direct effect of Stx2 in human trophoblast migration