title: Coxsackievirus preferentially replicates and induces cytopathic effects in undifferentiated neural progenitor cells.
abstract: Enteroviruses, including coxsackieviruses, exhibit significant tropism for the central nervous system, and these viruses are commonly associated with viral meningitis and encephalitis. Previously, we described the ability of coxsackievirus B3 (CVB3) to infect proliferating neuronal progenitor cells located in the neonatal subventricular zone and persist in the adult murine central nervous system (CNS). Here, we demonstrate that cultured murine neurospheres, which comprise neural stem cells and their progeny at different stages of development, were highly susceptible to CVB3 infection. Neurospheres, or neural progenitor and stem cells (NPSCs), isolated from neonatal C57BL/6 mice, supported high levels of infectious virus production and high viral protein expression levels following infection with a recombinant CVB3 expressing enhanced green fluorescent protein (eGFP) protein. Similarly, NPSCs isolated from neonatal actin-promoter-GFP transgenic mice (actin-GFP NPSCs) were highly susceptible to infection with a recombinant CVB3 expressing DsRed (Discosoma sp. red fluorescent protein). Both nestin-positive and NG2(+) progenitor cells within neurospheres were shown to preferentially express high levels of viral protein as soon as 24 h postinfection (p.i.). By day 3 p.i., viral protein expression and viral titers increased dramatically in NPSCs with resultant cytopathic effects (CPE) and eventual cell death. In contrast, reduced viral replication, lower levels of CPE, and diminished viral protein expression levels were observed in NPSCs differentiated for 5 or 16 days in the presence of fetal bovine serum (FBS). Despite the presence of CPE and high levels of cell death following early CVB3 infection, surviving neurospheres were readily observed and continued to express detectable levels of viral protein as long as 37 days after initial infection. Also, CVB3 infection of actin-GFP NPSCs increased the percentage of cells expressing neuronal class III β-tubulin following their differentiation in the presence of FBS. These results suggest that neural stem cells may be preferentially targeted by CVB3 and that neurogenic regions of the CNS may support persistent viral replication in the surviving host. In addition, normal progenitor cell differentiation may be altered in the host following infection.
authors.affiliation: Cell and Molecular Biology Joint Doctoral Program, Department of Biology, San Diego State University, 5500 Campanile Drive, San Diego, CA 92182-4614, USA.
authors.forename: Ginger
authors.initials: G
authors.lastname: Tsueng
authors.forename: Jenna M
authors.initials: JM
authors.lastname: Tabor-Godwin
authors.forename: Aparajita
authors.initials: A
authors.lastname: Gopal
authors.forename: Chelsea M
authors.initials: CM
authors.lastname: Ruller
authors.forename: Steven
authors.initials: S
authors.lastname: Deline
authors.forename: Naili
authors.initials: N
authors.lastname: An
authors.forename: Ricardo F
authors.initials: RF
authors.lastname: Frausto
authors.forename: Richard
authors.initials: R
authors.lastname: Milner
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Crocker
authors.forename: J Lindsay
authors.initials: JL
authors.lastname: Whitton
authors.forename: Ralph
authors.initials: R
authors.lastname: Feuer
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D003588:Cytopathogenic Effect, Viral; D029822:Enterovirus B, Human / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology* / Q000648:ultrastructure; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D058953:Neural Stem Cells / Q000166:cytology / Q000648:ultrastructure / Q000821:virology*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication; D000097573:Red Fluorescent Protein
chemical_list: D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D014764:Viral Proteins; D000097573:Red Fluorescent Protein; C485184:enhanced green fluorescent protein