A single intramuscular injection of recombinant plasmid DNA induces protective immunity and prevents Japanese encephalitis in mice
A single intramuscular injection of recombinant plasmid DNA induces protective immunity and prevents Japanese encephalitis in mice
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DOI：
10.1128/jvi.74.9.4244-4252.2000
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发表时间：
2000-05-01
影响因子：
5.4
通讯作者：
Davis, B
中科院分区：
文献类型：
--
作者：
Chang, GJJ;Hunt, AR;Davis, B
构建了含有日本脑炎病毒（JEV）前膜（prM）和包膜（E）基因的质粒载体，其在巨细胞病毒立即早期基因启动子的控制下表达prM和E蛋白。用该质粒载体（JE-4 B克隆）转化的COS-1细胞将JEV特异性细胞外颗粒（EP）分泌到培养基中。免疫后18个月，所有小鼠均能保持JEV特异性抗体。JEVAX在3周龄小鼠中诱导100%血清转化;然而，没有一只3日龄小鼠的酶联免疫吸附测定滴度高于1：400。用该DNA疫苗免疫的雌性小鼠产生的空斑减少中和抗体滴度在1：20和1：160之间，并且在用5，000 PFU的强毒株JEV SA 14腹膜内攻击后对其后代提供45至100%的被动保护。7周龄的成年小鼠在3日龄时接受了单剂量的JEV DNA疫苗，完全保护其免受50，000-PFU JEV腹膜内攻击。这些结果表明，在体外产生JEV EPs的重组质粒DNA是一种有效的疫苗。
Plasmid vectors containing Japanese encephalitis virus (JEV) premembrane (prM) and envelope (E) genes were constructed that expressed prM and E proteins under the control of a cytomegalovirus immediate-early gene promoter. COS-1 cells transformed with this plasmid vector (JE-4B clone) secreted JEV-specific extracellular particles (EPs) into the culture media. Groups of outbred ICR mice were given one or two doses of recombinant plasmid DNA or two doses of the commercial vaccine JEVAX, All mice that received one or two doses of DNA vaccine maintained JEV-specific antibodies 18 months after initial immunization. JEVAX induced 100% seroconversion in 3-week-old mice; however, none of the 3-day-old mice had enzyme-linked immunosorbent assay titers higher than 1:400. Female mice immunized with this DNA vaccine developed plaque reduction neutralization antibody titers of between 1:20 and 1:160 and provided 45 to 100% passive protection to their progeny following intraperitoneal challenge with 5,000 PFU of virulent JEV strain SA14. Seven-week-old adult mice that had received a single dose of JEV DNA vaccine when 3 days of age were completely protected from a 50,000-PFU JEV intraperitoneal challenge. These results demonstrate that a recombinant plasmid DNA which produced JEV EPs in vitro is an effective vaccine.