Transfection of prion protein gene suppresses coxsackievirus B3 replication in prion protein gene-deficient cells

• Published in: Journal of general virology Vol. 84; no. 12; pp. 3495 - 3502
• Main Authors: Nakamura, Yuko, Sakudo, Akikazu, Saeki, Keiichi, Kaneko, Tomomi, Matsumoto, Yoshitsugu, Toniolo, Antonio, Itohara, Shigeyoshi, Onodera, Takashi
• Format: Journal Article
• Language: English
• Published: England Soc General Microbiol 01.12.2003
• Subjects: Animals; Animals, Newborn; Apoptosis; Brain; Cells, Cultured; Coxsackievirus B3; Cytopathogenic Effect, Viral; Disease Models, Animal; Disease Susceptibility; Enterovirus B, Human - physiology; Enterovirus Infections - prevention & control; Enterovirus Infections - virology; Humans; Interferon Type I - biosynthesis; Mice; Mice, Inbred C57BL; Neurons - metabolism; Neurons - virology; Prions - biosynthesis; Prions - genetics; Transfection; Virus Replication
• ISSN: 0022-1317; 1465-2099
• DOI: 10.1099/vir.0.19222-0

1 Department of Molecular Immunology, School of Agricultural and Life Sciences, University of Tokyo, 1-1-1, Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan 2 Department of Clinical and Biological Sciences, University of Insubria, Varese, Italy 3 Laboratory for Behavioural Genetics, Brain Science Institute, RIKEN, Saitama, Japan Correspondence Takashi Onodera aonoder{at}mail.ecc.u-tokyo.ac.jp 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.