In vivo electroporation enhances the immunogenicity of hepatitis C virus nonstructural 3/4A DNA by increased local DNA uptake, protein expression, inflammation, and infiltration of CD3+ T cells
In vivo electroporation enhances the immunogenicity of hepatitis C virus nonstructural 3/4A DNA by increased local DNA uptake, protein expression, inflammation, and infiltration of CD3+ T cells
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DOI：
10.4049/jimmunol.179.7.4741
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发表时间：
2007-10-01
影响因子：
4.4
通讯作者：
Sallberg, Matti
中科院分区：
文献类型：
--
作者：
Ahlen, Gustaf;Soderholm, Jonas;Sallberg, Matti
体内电穿孔（EP）提高i.m. DNA疫苗的特点是使用丙型肝炎病毒非结构（NS）3/4A基因。在标准的i.m.注射有或没有体内EP的DNA，质粒水平在注射部位立即达到峰值，第一周下降4个对数。在体内EP没有促进质粒的持久性，并根据剂量，质粒被清除或几乎清除后60天。在体内成像和免疫组织化学显示，蛋白质表达仅限于注射部位，尽管在相邻的肌肉群中检测到显着水平的质粒。在体内，EP增加并延长了NS 3/4A蛋白表达水平以及注射部位CD 3（+）T细胞浸润的增加。这些因素很可能是NS 3/4A特异性抗体、CD 4（+）T细胞、CD 8（+）T细胞和γ-IFN产生的增强和扩大的加和作用。引发的CD 8（+）应答在体内是功能性的，导致瞬时转基因小鼠中丙型肝炎病毒NS 3/4A表达肝细胞的消除。总的来说，体内EP后注射部位的蛋白表达增强和炎症有助于体内功能性免疫应答的引发。这些局部效应很可能有助于确保在较大动物（包括兔子和人类）中测试疫苗时保持反应的强度和持续时间。因此，由体内EP介导的组合效应充当基于NS 3/4A的DNA疫苗的有效佐剂。
The mechanisms by which in vivo electroporation (EP) improves the potency of i.m. DNA vaccination were characterized by using the hepatitis C virus nonstructural (NS) 3/4A gene. Following a standard i.m. injection of DNA with or without in vivo EP, plasmid levels peaked immediately at the site of injection and decreased by 4 logs the first week. In vivo EP did not promote plasmid persistence and, depending on the dose, the plasmid was cleared or almost cleared after 60 days. In vivo imaging and immiumo-histochemistry revealed that protein expression was restricted to the injection site despite the detection of significant levels of plasmid in adjacent muscle groups. In vivo EP increased and prolonged NS3/4A protein expression levels as well as an increased infiltration of CD3(+) T cells at the injection site. These factors most likely additively contributed to the enhanced and broadened priming of NS3/4A-specific Abs, CD4(+) T cells, CD8(+) T cells, and gamma-IFN production. The primed CD8(+) responses were functional in vivo, resulting in elimination of hepatitis C virus NS3/4A-expressing liver cells in transiently transgenic mice. Collectively, the enhanced protein expression and inflammation at the injection site following in vivo EP contributed to the priming of in vivo functional immune responses. These localized effects most likely help to insure that the strength and duration of the responses are maintained when the vaccine is tested in larger animals, including rabbits and humans. Thus, the combined effects mediated by in vivo EP serves as a potent adjuvant for the NS3/4A-based DNA vaccine.