altering an artificial gagpolnef polyprotein and mode of env co-administration affects the immunogenicity of a clade c hiv dna vaccine改变一个人工gagpolnef多蛋白和env合并施打模式影响分支c艾滋病毒dna疫苗的免疫原性.pdfVIP

Altering an Artificial Gagpolnef Polyprotein and Mode of ENV Co-Administration Affects the Immunogenicity of a Clade C HIV DNA Vaccine ¨ 1. 1. 1¤a ¨ 1¤b ¨ 1 1,2 Katharina Bockl , Jens Wild , Simon Bredl , Kathrin Kindsmuller , Josef Kostler , Ralf Wagner * 1 Institute of Medical Microbiology, University of Regensburg, Regensburg, Germany, 2 Geneart AG/Life Technologies, Regensburg, Germany Abstract HIV-1 candidate vaccines expressing an artificial polyprotein comprising Gag, Pol and Nef (GPN) and a secreted envelope protein (Env) were shown in recent Phase I/II clinical trials to induce high levels of polyfunctional T cell responses; however, Env-specific responses clearly exceeded those against Gag. Here, we assess the impact of the GPN immunogen design and variations in the formulation and vaccination regimen of a combined GPN/Env DNA vaccine on the T cell responses against the various HIV proteins. Subtle modifications were introduced into the GPN gene to increase Gag expression, modify the expression ratio of Gag to PolNef and support budding of virus-like particles. I.m. administration of the various DNA constructs into BALB/c mice resulted in an up to 10-fold increase in Gag- and Pol-specific IFNc+ CD8+ T cells compared to GPN. Co-administering Env with Gag or GPN derivatives largely abrogated Gag-specific responses. Alterations in the molar ratio of the DNA vaccines and spatially or temporally separated administration induced more balanced T cell responses. Whereas forced co-expression of Gag and Env from one plasmid induced predominantly Env-specific T cells responses, deletion of the only H-2d T cell epitope in E