Characterization of a DNA vaccine expressing a human immunodeficiency virus-like particle.
Characterization of a DNA vaccine expressing a human immunodeficiency virus-like particle.
复制标题
表达人类免疫缺陷病毒样颗粒的 DNA 疫苗的表征。
DOI：
10.1016/j.virol.2004.07.009
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发表时间：
2004
期刊：
影响因子：
3.7
通讯作者：
Ross,TedM
中科院分区：
文献类型：
--
作者：
Young,KellyR;Smith,JamesM;Ross,TedM
一种理想的人类免疫缺陷病毒1型（HIV-1）疫苗很可能需要引发交叉反应性中和抗体和针对多种HIV-1抗原的强细胞介导免疫反应，以赋予对攻击的保护。在本研究中，构建了表达病毒调控的人类免疫缺陷病毒样颗粒（VLP）的DNA疫苗，以引发对多种HIV-1抗原的广谱免疫反应。利用从X4分离物中编码gag和pol基因产物的序列，以及从R5或R5X4分离物中编码tat、rev、vpu和env的序列，有效地制备了VLPs。整合酶、vpr、vif和nef基因被删除。此外，长末端重复序列（LTRs）被移除，VLP插入片段的转录被添加巨细胞病毒立即早期（CMV-IE）启动子驱动。构建了第二代VLP疫苗质粒，在VLP DNA中进行突变，以产生缺乏病毒逆转录酶和蛋白酶相关活性的颗粒。短暂转染DNA的灵长类细胞系有效地将VLP分泌到上清液中，上清液在蔗糖梯度内呈带状，密度与感染性病毒粒子相似。此外，这些颗粒在结合可溶性人CD4的颗粒表面结合Env。这些VLPs提供了一种安全有效的策略，将多个HIV-1抗原通过单个插入物表达，以模仿感染性病毒粒子的结构呈现给免疫系统。
An ideal human immunodeficiency virus type-1 (HIV-1) vaccine will most likely need to elicit cross-reactive neutralizing antibodies and a strong cell-mediated immune response against multiple HIV-1 antigens to confer protection against challenge. In this study, DNA vaccines were constructed to express virally regulated human immunodeficiency virus-like particles (VLP) to elicit broad-spectrum immune responses to multiple HIV-1 antigens. VLPs were efficiently produced using sequences encoding gag and pol gene products from an X4 isolate and sequences encoding for tat, rev, vpu, and env from R5 or R5X4 isolates. The integrase, vpr, vif, and nef genes were deleted. In addition, the long terminal repeats (LTRs) were removed and transcription of the VLP insert was driven by the addition of the cytomegalovirus immediate-early (CMV-IE) promoter. A second generation of VLP vaccine plasmids was constructed with mutations engineered into the VLP DNA to produce particles deficient in activities associated with viral reverse transcriptase and protease. Primate cell lines, transiently transfected with DNA, efficiently secreted VLP into the supernatant that banded within a sucrose gradient at densities similar to infectious virions. In addition, these particles incorporated Env on the particle surface that bound soluble human CD4. These VLPs provide a safe and efficient strategy for presenting multiple HIV-1 antigens, expressed from a single insert, to the immune system in a structure that mimics the infectious virion.