COXSACKIEVIRUS B INFECTION INDUCES APOPTOSIS IN HUMAN EMBRYONIC STEM CELLS

ROMORINI LEONARDO; SCASSA MARÍA ELIDA; BLUGUERMANN CAROLINA; RICHARDSON VIDELA GUILLERMO; QUESTA MARÍA; FERNANDEZ ESPINOZA DARIO DAMIAN; SEVLEVER GUSTAVO EMILIO; GOMEZ RICARDO M; MIRIUKA SANTIAGO GABRIEL

Congreso; 2011 9th Annual Meeting International Society for Stem Cell Research; 2011

ISSCR

Human embryonic stem cells (hESCs) are self-renewing pluripotent cells thatcan differentiate to a wide range of specialized cells. Group B Coxsackieviruses (CVBs) produce acute myocarditis and chronic dilated cardiomyopathy.We have previously shown that hESCs and differentiated contractileembryoid bodies express CVB receptors and that CVBs infection of thesecells causes a cytopathic effect compatible with cell death. The aim of thepresent work was to study if CVBs infection induces programmed cell deathin hESCs. hESCs HUES-5 (H5) and WA-01 (H1) and CVBs (1 and 3) wereused. Undifferentiated state of hESCs was validated by immunofluorescenceand RT-PCR of pluripotent markers (TRA1-60, TRA1-81, SSEA-4, Oct-4 andNanog). H1 and H5 cell viability (XTT assay) and apoptosis (DAPI staining,TUNEL, PARP cleavage and caspase-3-like activity assays) were measuredat different time points post-infection (pi). Cell viability decreased dependingon moi at 24hs CVB1 and CVB3 pi; DAPI and TUNEL positive apoptoticnuclei increased at 5 and 8 hours CVB3 pi, caspase-3-like activity peaks at1 hour CVB3 pi and PARP cleavage started at 5 hours CVB3 pi. Quantificationof anti-(Bcl-2, Bcl-XL) and pro-(Bax, Bad) apoptotic genes mRNA levelsby RT-Real Time PCR showed a decrease in Bcl-XL mRNA levels for bothH1 and H5 at 8hs CVB3 pi. However Bcl-2, Bcl-XL and Bax protein levelsdid not change at 5, 8, 16 and 24 hours CVB3 pi. Conclusions: Cytopathiceffect observed after hESCs infection with CVBs is compatible with an earlyinduction of apoptosis.