Human embryonic stem cells and derived contractile embryoid bodies are susceptible to Coxsakievirus B infection and respond to interferon Iβ treatment.
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 differentiation stages studied. Real-time PCR analysis showed that CAR mRNA levels were 3.4-fold higher in undifferentiated cells, while DAF transcript levels were 2.78-fold more abundant in differentiated cultures (P<0.05). All cell lines were susceptible to Coxsackievirus serotypes B1-5 infection as shown by RT-PCR detection of viral RNA, immunofluorescence detection of viral protein and infectivity titration of cell culture supernatants resulting in cell death. Supernatants infectivity titers 24-48 h post-infection ranged from 10⁵-10⁶ plaque forming units (PFU)/ml, the highest titers were detected in undifferentiated cells. Cell viability 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 24-48 h observation period, as detected by reduced infectivity titers in the supernatants and increased cell viability by a colorimetric assay, respectively. We propose human embryonic stem cell and derived contractile embryoid bodies as a valid model to study cardiac Coxsackievirus B infection.
Scassa
María E
ME
Laboratorio de Biología del Desarrollo Celular, FLENI, Buenos Aires, Argentina.
Jaquenod de Giusti
Carolina
C
Questa
María
M
Pretre
Gabriela
G
Richardson
Guillermo A Videla
GA
Bluguermann
Carolina
C
Romorini
Leonardo
L
Ferrer
María F
MF
Sevlever
Gustavo E
GE
Miriuka
Santiago G
SG
Gómez
Ricardo M
RM
D002460:Cell Line; D003384:Coxsackievirus Infections / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D058732:Embryoid Bodies / Q000187:drug effects / Q000821:virology*; D053595:Embryonic Stem Cells / Q000187:drug effects / Q000821:virology*; D029822:Enterovirus B, Human / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D016899:Interferon-beta / Q000494:pharmacology*; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects
D011991:Receptors, Virus; D016899:Interferon-beta