J Bacteriol Virol.  2007 Mar;37(1):23-30. 10.4167/jbv.2007.37.1.23.

Development of Neutralization Assay using Murine Leukemia Virus (MuLV) Pseudotyped with Japanese encephalitis Virus (JEV) env Gene

Affiliations
  • 1Department of Animal Biotechnology, College of Animal Bioscience & Technology, Konkuk University, Seoul, 143-701, Korea. kimera@konkuk.ac.kr
  • 2Biological Diagnostic Products Team Biologics Headquarters #231 Jinheungno Eunpyung-Gu, Seoul,122-704, Korea.
  • 3Department of Biotechnology, The Catholic University of Korea, 43-1 Yeokgok Dong, Wonmi-Ku, Bucheon, 420-743, Korea.

Abstract

The envelope (E) glycoprotein of JEV is the major antigen to elicit neutralizing antibody (NAb) against JEV infection. In order to develop a rapid and safe neutralization assay system for evaluation of the JEV vaccine strains, we constructed JEV-pseudotyped viruses with JEV env genes (Nakayama-NIH, Beijing-1). The titers of JEV-pseudotyped viruses with NK and BJ strains were 4.0x10(4) IFU/ml and 1.3x10(5) IFU/ml in Vero cell cultures, respectively. We have analyzed the neutralization activity of immunized mouse sera with JEV-NK and JEV-BJ pseudotyped viruses. The neutralizing antibody titers of NK and BJ (50% reduction of virus) were about 1:10,000 at each immunized sera. Compared with conventional plaque reduction neutralization test (PRNT), the method using JEV-pseudotyped virus has desirable advantages such as more rapid, easier, and non-biohazardous. This neutralization assay system might be useful to evaluate NAb activity against JEV vaccine strains or vaccine candidates.

Keyword

JEV; Envelope (E); Pseudotyped virus; Neutralization

MeSH Terms

Animals
Antibodies, Neutralizing
Asian Continental Ancestry Group*
Encephalitis Virus, Japanese*
Encephalitis, Japanese*
Genes, env*
Glycoproteins
Humans
Leukemia Virus, Murine*
Mice
Neutralization Tests
Vero Cells
Antibodies, Neutralizing
Glycoproteins
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