Human embryonic stem cells and derived contractile embryoid bodies are susceptible to Coxsackievirus B infection and respond to interferon I-β treatment

SCASSA, MARÍA E.; JAQUENOD DE GIUSTI, CAROLINA; QUESTA, MARÍA; PRETRE, GABRIELA; VIDELA RICHARDSON, GUILLERMO; BLUGUERMANN, CAROLINA; ROMORINI, LEONARDO; FERRER, MARÍA F.; SEVLEVER, GUSTAVO E.; MIRIUKA, SANTIAGO G.; GÓMEZ, RICARDO M.

STEM CELL RESEARCH

ELSEVIER SCIENCE BV

Año: 2011 vol. 6 p. 13 - 22

1873-5061

We studied the susceptibility of human embryonic stem cells and derived contractile embryoid bodies from WAO9,HUES-5 and HUES-16 cell lines to Coxsackievirus B infection. After validating stem cell-like properties and cardiac phenotype,Coxsackievirus B receptors CAR and DAF, as well as type I interferon receptors were detected in all cell lines and differentiationstages studied. Real-time PCR analysis showed that CAR mRNA levels were 3.4-fold higher in undifferentiated cells, while DAFtranscript levels were 2.78-fold more abundant in differentiated cultures (Pb0.05). All cell lines were susceptible toCoxsackievirus serotypes B1-5 infection as shown by RT-PCR detection of viral RNA, immunofluorescence detection of viralprotein and infectivity titration of cell culture supernatants resulting in cell death. Supernatants infectivity titers 24-48 h postinfectionranged from 105-106 plaque forming units (PFU)/ml, the highest titers were detected in undifferentiated cells. Cellviability detected by a colorimetric assay, showed inverse correlation with infectivity titers of cell culture supernatants.Treatment with 100 U of interferon Iβ significantly reduced viral replication and associated cell death during a 24?48h observation period, as detected by reduced infectivity titers in the supernatants and increased cell viability by a colorimetricassay, respectively. We propose human embryonic stem cell and derived contractile embryoid bodies as a valid model to studycardiac Coxsackievirus B infection.