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J Virol, June 1998, p. 4925-4930, Vol. 72, No. 6
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Induction of Protective Immunity against Japanese Encephalitis in Mice by Immunization with a Plasmid Encoding Japanese Encephalitis Virus Premembrane and Envelope Genes

Eiji Konishi,1,2,* Masaoki Yamaoka,1 Khin-Sane-Win,1 Ichiro Kurane,3 and Peter W. Mason4

Department of Medical Zoology, Kobe University School of Medicine, Kobe 650,1 Department of Health Sciences, Kobe University School of Medicine, Kobe 654-01,2 and Department of Microbiology, Kinki University School of Medicine, Osaka-Sayama 589,3 Japan, and Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, New York 119444

Received 5 January 1998/Accepted 24 February 1998

A DNA vaccine plasmid containing the Japanese encephalitis (JE) virus premembrane (prM) and envelope (E) genes (designated pcDNA3JEME) was evaluated for immunogenicity and protective efficacy in mice. Two immunizations of 4-week-old female ICR mice with pcDNA3JEME by intramuscular or intradermal injections at a dose of 10 or 100 µg per mouse elicited neutralizing (NEUT) antibodies at titers of 1:10 to 1:20 (90% plaque reduction), and all immunized mice survived a challenge with 10,000 50% lethal doses of the P3 strain of JE virus. A single immunization with 100 µg of pcDNA3JEME did not elicit detectable NEUT antibodies but induced protective immunity. Spleen cells obtained from BALB/c mice immunized once with 10 or 100 µg of pcDNA3JEME contained JE virus-specific memory cytotoxic T lymphocytes (CTLs). BALB/c mice maintained detectable levels of memory B cells and CTLs for at least 6 months after one immunization with pcDNA3JEME at a dose of 100 µg. The CTLs induced in BALB/c mice immunized twice with 100 µg of pcDNA3JEME were CD8 positive and recognized mainly the envelope protein. These results indicate that pcDNA3JEME has the ability to induce a protective immune response which includes JE virus-specific antibodies and CTLs.

* Corresponding author. Mailing address: Department of Health Sciences, Kobe University School of Medicine, 7-10-2 Tomogaoka, Suma-ku, Kobe 654-01, Japan. Phone: 81-78-796-4594. Fax: 81-78-796-4594. E-mail: ekon{at}ams.kobe-u.ac.jp.

J Virol, June 1998, p. 4925-4930, Vol. 72, No. 6
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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