Coxsackievirus b infection induces apoptosis in human embryonic stem cells.

LEONARDO ROMORINI; MARÍA ELIDA SCASSA; CAROLINA JAQUENOD DE GIUSTI; CAROLINA BLUGUERMANN; GUILLERMO VIDELA RICHARDSON; DARÍO FERNÁNDEZ ESPINOSA; RICARDO GÓMEZ; SANTIAGO G. MIRIUKA

Puerto Madryn

Congreso; SAIB; 2010

Human embryonic stem cells (hESCs) are self-renewing pluripotent cells that can differentiate to a wide range of specialized cells. Group B Coxsackie viruses (CVBs) produce acute myocarditis and chronic dilated cardiomyopathy. We have shown that hESCs and differentiated contractile embryoid bodies express CVB receptors and that CVBs infection of these cells cause a cytopathic effect compatible with cell death. The aim of the present work was to study if CVBs infection induces programmed cell death in hESCs. hESCs HUES-5 (H5) and WA-01 (H1) and CVBs (1 and 3) were used. Undifferentiated state of hESCs was validated by immunofluorescense 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 caspasa-3-like activity assays) were measured at different time point post-infection (pi). Cell viability decreased moi dependently at 24hs CVB1 and CVB3 pi; DAPI and TUNEL positive apoptotic nuclei increased at 5 and 8 hours CVB3 pi and caspase-3-like activity peaks at 1 hour CVB3 pi. Quantification of anti-(Bcl-2, Bcl-XL) and pro-(Bax, Bad) apopotic genes mRNA levels by RT-Real Time PCR showed a decrease in Bcl-XL mRNA levels for both H1 and H5 at 8hs CVB3 pi Conclusions: cytopathic effect observed after hESCs infection with CVBs is compatible with an induction of apoptotis.