COXSACKIEVIRUS B INFECTION INDUCES APOPTOSIS IN HUMAN EMBRYONIC STEM CELLS

ROMORINI LEONARDO; SCASSA MARÍA ELIDA; JAQUENOUD DE GUISTI CAROLINA; BLUGUERMANN CAROLINA; RICHARDSON VIDELA GUILLERMO; FERNANDEZ ESPINOZA DARIO DAMIAN; GOMEZ RICARDO M; MIRIUKA SANTIAGO GABRIEL

Pto Madryn

Congreso; XLVI Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2010

.Human embryonic stem cells (hESCs) are self-renewing pluripotentcells that can differentiate to a wide range of specialized cells. GroupB Coxsackie viruses (CVBs) produce acute myocarditis and chronicdilated cardiomyopathy. We have shown that hESCs anddifferentiated contractile embryoid bodies express CVB receptorsand that CVBs infection of these cells causes a cytopathic effectcompatible with cell death. The aim of the present work was to studyif CVBs infection induces programmed cell death in hESCs. hESCsHUES-5 (H5) and WA-01 (H1) and CVBs (1 and 3) were used.Undifferentiated state of hESCs was validated byimmunofluorescence and RT-PCR of pluripotent markers (TRA1-60, TRA1-81, SSEA-4, Oct-4 and Nanog). H1 and H5 cell viability(XTT assay) and apoptosis (DAPI staining, TUNEL and caspase-3-like activity assays) were measured at different time points postinfection(pi). Cell viability decreased moi dependently at 24hsCVB1 and CVB3 pi; DAPI and TUNEL positive apoptotic nucleiincreased at 5 and 8 hours CVB3 pi and caspase-3-like activity peaksat 1 hour CVB3 pi. Quantification of anti-(Bcl-2, Bcl-XL) and pro-(Bax, Bad) apoptotic genes mRNA levels by RT-Real Time PCRshowed a decrease in Bcl-XL mRNA levels for both H1 and H5 at8hs CVB3 pi. Conclusions: cytopathic effect observed after hESCsinfection with CVBs is compatible with an induction of apoptosis