NOVEL ADENOVIRUS-VECTORED HIV VACCINE THAT KNOCKS THE SOCS1 OFF DENDRITIC CELLS

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。次级项目的主要支助 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 表示子项目使用的中心基础设施的估计数量， NCRR赠款不直接向子项目或子项目工作人员提供资金。 抗原呈递细胞（如DC）在建立和维持针对HIV感染的免疫应答中起关键作用; DC是刺激免疫信号传导的疫苗的关键靶标。 我们在本资助期内取得了以下进展： + 开发了共表达（i）SOCS 1 siRNA和（ii）报告基因绿色荧光蛋白（GFP）或抗原基因HIV-1 gag的腺病毒载体组。 + 测试了SOCS 1 siRNA对树突状细胞表型的影响。 未观察到对树突状细胞成熟的负面影响。 + 开发了一种Western印迹方法，用于检测SOCS 1 siRNA表达对INF-γ刺激后STAT 1磷酸化的影响，作为SOCS 1 siRNA沉默的替代测量。 这些实验的结果表明，我们的SOCS 1 siRNA表达载体增强STAT 1信号通路，并支持我们的假设。 意义：开发一种新型腺病毒载体平台，旨在直接调节疫苗诱导的免疫应答。 此外，迄今为止的工作表明，在诱导免疫应答期间减弱SOCS 1的下调作用将有助于显著提高疫苗效力。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Antigen-presenting cells, such as DCs, play a critical role in the establishment and maintenance of the immune responses against HIV infection; DCs are key targets for vaccines that stimulate immune signaling. We have made the following progress during this funding period: + Developed panel of adenovirus-based vectors that co-express (i) SOCS1 siRNA and (ii) the reporter gene Green Fluorescent Protein (GFP) or antigen gene HIV-1 gag. + Have tested effects of SOCS1 siRNA on dendritic cell phenotype. No negative effects on dendritic cell maturation were observed. + Developed a Western blot approach for detecting the effects of SOCS1 siRNA expression on the phosphorylation of STAT1 following INF-gamma stimulation as a surrogate measurement of SOCS1 siRNA-silencing. The results from these experiments suggest that our SOCS1 siRNA-expressing vectors enhance the STAT1 signaling pathway and support our hypothesis. Significance: Development of a novel adenovirus vector platform that is designed to directly modulate the immune responses induced by the vaccine. Moreover, the work thus far suggests that attenuating the down-regulatory role of SOCS1 during the induction of immune responses will serve to significantly improve vaccine efficacy.