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Journal of Virology, July 1999, p. 5527-5534, Vol. 73, No. 7
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

The Anamnestic Neutralizing Antibody Response Is Critical for Protection of Mice from Challenge following Vaccination with a Plasmid Encoding the Japanese Encephalitis Virus Premembrane and Envelope Genes

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

Department of Health Sciences, Kobe University School of Medicine, Kobe 654-0142,1 Department of Medical Zoology, Kobe University School of Medicine, Kobe 650-0017,2 and Department of Microbiology, Kinki University School of Medicine, Osaka-sayama 589-8511,3 Japan, and Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, New York 119444

Received 19 November 1998/Accepted 30 March 1999

For Japanese encephalitis (JE), we previously reported that recombinant vaccine-induced protection from disease does not prevent challenge virus replication in mice. Moreover, DNA vaccines for JE can provide protection from high challenge doses in the absence of detectable prechallenge neutralizing antibodies. In the present study, we evaluated the role of postchallenge immune responses in determining the outcome of JE virus infection, using mice immunized with a plasmid, pcDNA3JEME, encoding the JE virus premembrane (prM) and envelope (E) coding regions. In the first experiment, 10 mice were vaccinated once (five animals) or twice (remainder) with 100 µg of pcDNA3JEME. All of these mice showed low (6 of 10) or undetectable (4 of 10) levels of neutralizing antibodies. Interestingly, eight of these animals showed a rapid rise in neutralizing antibody following challenge with 10,000 50% lethal doses of JE virus and survived for 21 days, whereas only one of the two remaining animals survived. No unimmunized animals exhibited a rise of neutralizing antibody or survived challenge. Levels of JE virus-specific immunoglobulin M class antibodies were elevated following challenge in half of the unimmunized mice and in the single pcDNA3JEME-immunized mouse that died. In the second experiment, JE virus-specific primary cytotoxic T-lymphocyte (CTL) activity was detected in BALB/c mice immunized once with 100 µg of pcDNA3JEME 4 days after challenge, indicating a strong postchallenge recall of CTLs. In the third experiment, evaluation of induction of CTLs and antibody activity by plasmids containing portions of the prM/E cassette demonstrated that induction of CTL responses alone were not sufficient to prevent death. Finally, we showed that antibody obtained from pcDNA3JEME-immunized mice 4 days following challenge could partially protect recipient mice from lethal challenge. Taken together, these results indicate that neutralizing antibody produced following challenge provides the critical protective component in pcDNA3JEME-vaccinated mice.

* Corresponding author. Mailing address: Department of Health Sciences, Kobe University School of Medicine, 7-10-2 Tomogaoka, Suma-ku, Kobe 654-0142, Japan. Phone and Fax: 81-78-796-4594. E-mail: ekon{at}ams.kobe-u.ac.jp.

dagger Present address: Hyogo Prefectural Institute of Public Health, Kobe 652-0032, Japan.

Dagger Present address: Department of Virology 1, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.

§ Present address: Department of Neurology, Kinki University School of Medicine, Osaka-sayama 589-8511, Japan.

Journal of Virology, July 1999, p. 5527-5534, Vol. 73, No. 7
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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