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Journal of Virology, April 1999, p. 3095-3101, Vol. 73, No. 4
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Yellow Fever/Japanese Encephalitis Chimeric Viruses: Construction and Biological Properties

Thomas J. Chambers,1,* Ann Nestorowicz,2 Peter W. Mason,3 and Charles M. Rice4

Department of Molecular Microbiology and Immunology, St. Louis University Health Sciences Center, St. Louis, Missouri 631041; Endocrine Division, Lilly Research Laboratories, Indianapolis, Indiana 462852; USDA Agricultural Research Service, PIADC, Greenport, New York 119443; and Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 631104

Received 4 November 1998/Accepted 4 January 1999

A system has been developed for generating chimeric yellow fever/Japanese encephalitis (YF/JE) viruses from cDNA templates encoding the structural proteins prM and E of JE virus within the backbone of a molecular clone of the YF17D strain. Chimeric viruses incorporating the proteins of two JE strains, SA14-14-2 (human vaccine strain) and JE Nakayama (JE-N [virulent mouse brain-passaged strain]), were studied in cell culture and laboratory mice. The JE envelope protein (E) retained antigenic and biological properties when expressed with its prM protein together with the YF capsid; however, viable chimeric viruses incorporating the entire JE structural region (C-prM-E) could not be obtained. YF/JE(prM-E) chimeric viruses grew efficiently in cells of vertebrate or mosquito origin compared to the parental viruses. The YF/JE SA14-14-2 virus was unable to kill young adult mice by intracerebral challenge, even at doses of 106 PFU. In contrast, the YF/JE-N virus was neurovirulent, but the phenotype resembled parental YF virus rather than JE-N. Ten predicted amino acid differences distinguish the JE E proteins of the two chimeric viruses, therefore implicating one or more residues as virus-specific determinants of mouse neurovirulence in this chimeric system. This study indicates the feasibility of expressing protective antigens of JE virus in the context of a live, attenuated flavivirus vaccine strain (YF17D) and also establishes a genetic system for investigating the molecular basis for neurovirulence determinants encoded within the JE E protein.

* Corresponding author. Mailing address: Department of Molecular Microbiology and Immunology, St. Louis University Health Sciences Center, 1402 South Grand Blvd., St. Louis, MO 63104. Phone: (314) 577-8447. Fax: (314) 773-3403. E-mail: chambetj{at}wpogate.slu.edu.

Journal of Virology, April 1999, p. 3095-3101, Vol. 73, No. 4
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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