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|標題:||Evaluation of Genotype III Vaccine against Newly
Emerged Genotype I and Development of Genotype I
Vaccine Candidate to Improve Vaccine Potency against
Genotype I and III of Japanese Encephalitis Viruses
評估第三型基因型疫苗對抗新興第一型基因型及開發 第一型基因型候選疫苗以提升疫苗對抗第一型和第三 型基因型日本腦炎病毒之效力
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Genotype I (GI) Japanese encephalitis virus (JEV), which has replaced genotype III (GIII) virus, has become the dominant circulating virus in Asia. Vaccination was the
most effective method to control JEV transmission, but currently only GIII JEV-derived vaccines are available for human and animal use. Based on molecular survey data of mosquitoes, GI virus was first identified in Taiwan during 2008 and became the only circulating genotype within one year. In Taiwan, annual vaccination of sows and the compulsory vaccination of children has been implemented using GIII JEV-derived vaccines. Thus, the potential impact(s) of and solution(s) to the genotype replacement event in terms of JEV vaccines were evaluated in this study.
Initially, serum samples from vaccinated pigs and humans were collected in order to assess the potency of the live-attenuated GIII at222 virus vaccine and the
formalin-inactivated GIII Nakayama virus vaccine, respectively.
A reduced seropositivity rate and neutralizing activity against GI wild-type (WT) TC2009-1 virus were observed in immunized pigs as well as in vaccinated children as compared to against the vaccine strain or against the GIII WT CJN virus.
Before developing a GI-derived vaccine in order to enhance GI-specific neutralizing activity, it was important to investigate the antigenic and immunogenic
impact on JEV of formalin inactivation in order to help with choosing the type of vaccine. We found a decreased binding activity of the T16 murine monoclonal antibody
(mAb) against commercial formalin-inactivated Nakayama virus vaccines (FICV), and the formalin-treated JEV (FIV-JEV) Nakayama, SA14-14-2, T1P1, and YL2009-4 strains, when compared to untreated JE viruses (IV-JEV). The contribution of the neutralizing activity of IgG antibodies elicited by the residues 329/331/389 epitope in domain III of E protein (EDIII 329/331/389), which is recognized by T16 mAb, was found to be higher among mice immunized with formalin-untreated Nakayama virus than among mice immunized with formalin-treated virus or the commercial JEV vaccine. We concluded that the formalin-inactivation process introduces antigenic modification(s) of EDIII of JEV and distorts the profile of vaccine-induced neutralizing antibodies after the formalin-inactivated vaccine.
Based on the above results, we utilized a virus-like particle (VLP) expression plasmid to construct a GI-derived JEV vaccine and established JEV mutant
VLP-expression plasmids that encoded fusion peptide mutants in order to reduce the immunogenicity of low-neutralizing or non-neutralizing epitopes in the fusion peptide(FP). Four JEV DNA vaccines were evaluated as part of this study; these consisted of plasmids expressing the GI WT YL2009-4 virus VLP (pJGI WT), the GIII WT SA14
virus VLP (pJGIII WT), the GI FP mutant VLP (pJGI GKD), and the GIII FP mutant VLP (pJGIII GKD). The geometric mean titer (GMT) of the PRNT50 against the GI and
GIII viruses among pJGI WT immunized mice was higher than among pJGIII WT immunized mice; however, it was significantly reduced among pJGI GKD and pJGIII GKD immunized mice. In addition, only 43% (3/7) of the pJGIII WT immunized mice were able to survive challenge with the GI and GIII JEVs; in contrast, 100% (12/12) and 92% (11/12) of pJGI WT immunized mice survived GI and GIII JEVs infection,respectively. Among the JEV WT and mutant VLP-expression plasmids, pJGI WT was the most potent vaccine in terms of evoking cross-neutralizing antibodies and protecting mice from GI and GIII JEVs infection.
This study included a serosurvey of GI JEV infection in pigs together with an analysis of the vaccine efficacies of inactivated vaccine and of live-attenuated vaccine
against GI WT JEVs, which are the first time such investigations have been carried out in Taiwan. One important concern that arose from these results was the poor vaccine potency of the commercial GIII vaccine when challenged with GI WT JEVs and future monitoring of JEV activity in pigs and mosquitoes is important. In terms of a future JEV vaccine, there is a need to search for an antigenically stable inactivation method that can
be used to produce an effective inactivated vaccine and it will also be very valuable to evaluate any developed GI virus-derived vaccine in pigs as well as humans.
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