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Journal of Virology, April 2003, p. 4248-4260, Vol. 77, No. 7
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.7.4248-4260.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Inoculation of Plasmids Encoding Japanese Encephalitis Virus PrM-E Proteins with Colloidal Gold Elicits a Protective Immune Response in BALB/c Mice

Zijiang Zhao,1,2 Takaji Wakita,1 and Kotaro Yasui1*

Department of Microbiology and Immunology, Tokyo Metropolitan Institute for Neuroscience, Tokyo 183-8526, Japan,1 Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052, People's Republic of China2

Received 26 July 2002/ Accepted 10 January 2003

We established a simple and effective method for DNA immunization against Japanese encephalitis virus (JEV) infection with plasmids encoding the viral PrM and E proteins and colloidal gold. Inoculation of plasmids mixed with colloidal gold induced the production of specific anti-JEV antibodies and a protective response against JEV challenge in BALB/c mice. When we compared the efficacy of different inoculation routes, the intravenous and intradermal inoculation routes were found to elicit stronger and more sustained neutralizing immune responses than intramuscular or intraperitoneal injection. After being inoculated twice, mice were found to resist challenge with 100,000 times the 50% lethal dose (LD50) of JEV (Beijing-1 strain) even when immunized with a relatively small dose of 0.5 µg of plasmid DNA. Protective passive immunity was also observed in SCID mice following transfer of splenocytes or serum from plasmid DNA- and colloidal gold-immunized BALB/c mice. The SCID mice resisted challenge with 100 times the LD50 of JEV. Analysis of histological sections detected expression of proteins encoded by plasmid DNA in the tissues of intravenously, intradermally, and intramuscularly inoculated mice 3 days after inoculation. DNA immunization with colloidal gold elicited encoded protein expression in splenocytes and might enhance immune responses in intravenously inoculated mice. This approach could be exploited to develop a novel DNA vaccine.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Tokyo Metropolitan Institute for Neuroscience, 2-6 Musashidai, Fuchu-shi, Tokyo 183-8526, Japan. Phone: (81) 42 325 3881, ext. 4416. Fax: (81) 42 321 8678. E-mail: yasui{at}tmin.ac.jp.

Journal of Virology, April 2003, p. 4248-4260, Vol. 77, No. 7
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.7.4248-4260.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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