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Designing and engineering of DNA-vaccine construction encoding multiple CTL-epitopes of major HIV-1 antigens

机译：设计和工程化编码主要HIV-1抗原的多个CTL表位的DNA疫苗构建

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A synthetic T cell immunogen (TCI) has been designed as a candidate DNA-based vaccine against Human immunodeficiency virus (HIV)-1 using cytotoxic T lymphocytes (CD8(+) CTL) and T-helper lymphocytes (CD4(+) Th) epitopes retrieved from the Los Alamos HIV Molecular Immunology Database. The protein 392 amino acids in length contains about eighty CTL-epitopes, many of which are overlapping and are totally restricted by ten different HLA class I molecules. To be able to detect CTL responses induced by a DNA vaccine in experimental animals, additional epitopes, restricted by mouse and Macaque rhesus major histocompatibility complex (MHC) class I molecules, were included in the target immunogen. The gene encoding the TO protein was assembled, cloned into vector plasmids and expressed in a prokaryotic and a eukaryotic system. The presence of HIV-1 protein fragments in the immunogen structure was ascertained by ELISA and immunoblotting using panels of HIV-1-positive sera and monoclonal antibodies to p24. It has been demonstrated that DNA vaccine can induce both specific T cell responses (CTL and blast transformation) and specific antibodies in mice immunized with pcDNA-TCI
机译：使用细胞毒性T淋巴细胞（CD8（+）CTL）和T辅助淋巴细胞（CD4（+）Th）设计了合成的T细胞免疫原（TCI）作为抗人免疫缺陷病毒（HIV）-1的基于DNA的候选疫苗。从洛斯阿拉莫斯HIV分子免疫学数据库中检索到的抗原决定簇。长度为392个氨基酸的蛋白质包含约80个CTL表位，其中许多表位重叠并且完全受十种不同的I类HLA分子限制。为了能够在实验动物中检测到由DNA疫苗诱导的CTL反应，目标免疫原中包括了受小鼠和猕猴主要组织相容性复合体（MHC）I类分子限制的其他表位。组装编码TO蛋白的基因，将其克隆到载体质粒中，并在原核和真核系统中表达。通过ELISA和使用HIV-1阳性血清和针对p24的单克隆抗体的免疫印迹法确定免疫原结构中HIV-1蛋白片段的存在。已经证明DNA疫苗可以在用pcDNA-TCI免疫的小鼠中诱导特异性T细胞应答（CTL和胚细胞转化）和特异性抗体

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