Inoculation of Plasmids Encoding Japanese Encephalitis Virus PrM-E Proteins with Colloidal Gold Elicits a Protective Immune Response in BALB/c Mice
Inoculation of Plasmids Encoding Japanese Encephalitis Virus PrM-E Proteins with Colloidal Gold Elicits a Protective Immune Response in BALB/c Mice
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DOI：
10.1128/jvi.77.7.4248-4260.2003
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发表时间：
2003-04
影响因子：
5.4
通讯作者：
Zijiang Zhao;T. Wakita;K. Yasui
中科院分区：
文献类型：
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作者：
Zijiang Zhao;T. Wakita;K. Yasui
摘要利用编码乙型脑炎病毒PrM和E蛋白的质粒和胶体金建立了一种简单有效的DNA免疫方法。将质粒与胶体金混合接种BALB/c小鼠，可诱导小鼠产生特异性抗JEV抗体，并产生保护性应答。当我们比较不同接种途径的效力时，发现静脉内和皮内接种途径比肌肉内或腹膜内注射引起更强和更持续的中和免疫应答。两次接种后，即使用0.5 μg质粒DNA免疫小鼠，也能抵抗100，000倍于50%致死量（LD_（50））的JEV（北京1株）攻击。从质粒DNA和胶体金免疫的BALB/c小鼠转移脾细胞或血清后，在SCID小鼠中也观察到保护性被动免疫。SCID小鼠抵抗100倍于JEV LD 50的攻击。组织切片分析检测到由质粒DNA编码的蛋白质在接种后3天的静脉内、皮内和肌内接种的小鼠的组织中的表达。胶体金DNA免疫可诱导脾细胞表达编码蛋白，并可增强静脉接种小鼠的免疫应答。这种方法可以用来开发一种新的DNA疫苗。
ABSTRACT 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.