Transfection of prion protein gene suppresses coxsackievirus B3 replication in prion protein gene-deficient cells.
The susceptibility of prion protein gene (Prnp)-null cells to coxsackievirus B3 (CVB3) was investigated. Primary cultures of murine Prnp(-/-) brain cells were more sensitive to CVBs than corresponding cells from wild-type mice. The viral susceptibility of a Prnp-null cell line (HpL3-4) derived from the murine hippocampus was compared with that of two established cell lines (HeLa and HEp-2) that are widely employed for CVB3 studies. After infection with CVB3, HpL3-4 cells showed a very rapid and complete cytopathic effect (CPE). CPE developed earlier and viruses replicated at higher titres in HpL3-4 cells compared with HeLa and HEp-2 cells. Under a semi-solid medium, plaques developed rapidly in CVB3-infected HpL3-4 cells. To confirm the effect of Prnp on virus infection, a Prnp(-/-) cell line and a Prnp-transfected neuronal cell line were analysed. The replication and release of infectious particles of CVB3 in Prnp(-/-) cells were significantly more effective than those of the Prnp-transfected cell line. Levels of type I interferon (IFN) after CVB3 infection were higher in the Prnp-transfected cell line than in Prnp(-/-) cells, whereas apoptotic cells were more obvious in the Prnp(-/-) cells than in those of the Prnp-transfected cell line. These findings suggest that the absence of Prnp retards the induction of CVB3-induced IFNs, resulting in an enhanced CVB3 production and apoptotic cell death. Furthermore, our data indicate that the HpL3-4 cell line may provide a novel and sensitive system for isolation of CVB3 from clinical specimens.
Nakamura
Yuko
Y
Department of Molecular Immunology, School of Agricultural and Life Sciences, University of Tokyo, 1-1-1, Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
Sakudo
Akikazu
A
Department of Molecular Immunology, School of Agricultural and Life Sciences, University of Tokyo, 1-1-1, Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
Saeki
Keiichi
K
Department of Molecular Immunology, School of Agricultural and Life Sciences, University of Tokyo, 1-1-1, Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
Kaneko
Tomomi
T
Department of Molecular Immunology, School of Agricultural and Life Sciences, University of Tokyo, 1-1-1, Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
Matsumoto
Yoshitsugu
Y
Department of Molecular Immunology, School of Agricultural and Life Sciences, University of Tokyo, 1-1-1, Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
Toniolo
Antonio
A
Department of Clinical and Biological Sciences, University of Insubria, Varese, Italy.
Itohara
Shigeyoshi
S
Laboratory for Behavioural Genetics, Brain Science Institute, RIKEN, Saitama, Japan.
Onodera
Takashi
T
Department of Molecular Immunology, School of Agricultural and Life Sciences, University of Tokyo, 1-1-1, Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis; D001921:Brain; D002478:Cells, Cultured; D003588:Cytopathogenic Effect, Viral; D004195:Disease Models, Animal; D004198:Disease Susceptibility; D029822:Enterovirus B, Human / Q000502:physiology*; D004769:Enterovirus Infections / Q000517:prevention & control / Q000821:virology*; D006801:Humans; D007370:Interferon Type I / Q000096:biosynthesis; D051379:Mice; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000378:metabolism / Q000821:virology; D011328:Prions / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection; D014779:Virus Replication
D007370:Interferon Type I; D011328:Prions