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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 旨在通过修改人类免疫缺陷病毒-1(HIV)抗原和使用各种递送系统和佐剂来刺激免疫反应的努力迄今未能生产出预防初级感染的艾滋病毒疫苗，这突显了继续开发替代和有效的候选疫苗的必要性。我们的项目是基于这样的发现，即细胞因子信号转导抑制因子1(SOCS1)蛋白通过限制Janus激活的信号转导和转录激活因子以及Toll样受体信号通路而起到抗原提呈衰减器的作用。我们项目的目的是： 1)腺病毒介导的SOCS1 siRNA抑制DC SOCS1表达 2)确定SOCS1消音对DC功能的影响(S 3)评估SOCS1沉默诱导的HIV特异性免疫应答 腺病毒载体疫苗 在报告所述期间，我们： 开发了一组基于腺病毒的载体，共表达(I)SOCS1 siRNA和(Ii)报告基因绿色荧光蛋白(GFP)或抗原基因HIV-1Gag。 我们检测了SOCS1 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。 建立了一种Western印迹方法来检测在INF-Gamma刺激后SOCS1 siRNA表达对STAT1磷酸化的影响，作为SOCS1 siRNA沉默的替代测量。这些实验的结果表明，我们的SOCS1 siRNA表达载体增强了STAT1信号通路，并支持我们的假设。 在此期间取得的进展的意义主要与开发一种新型的腺病毒载体平台有关，该平台旨在直接调节疫苗诱导的免疫反应。此外，到目前为止的工作表明，通过减弱SOCS1在诱导免疫反应过程中的下调作用，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. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Efforts aimed at stimulating immune responses by modifying Human Immunodeficiency virus-1 (HIV) antigens and using a variety of delivery systems and adjuvants have so far failed to produce an HIV vaccine that protects against primary infections, highlighting the need for the continued development of alternative and effective vaccine candidates. Our project is based on findings that the suppressor of cytokine signaling 1 (SOCS1) protein functions as an antigen-presentation attenuator by restricting the Janus-activated kinasesignal transducers and activators of transcription and Toll-like receptor-signaling pathways. The purpose of our project was to: 1) Inhibit DC SOCS1 expression with Ad-mediated delivery of SOCS1 siRNA 2) Determine the effect(s) of SOCS1-silencing on DC function 3) Evaluate the HIV-specific immunes responses induced by SOCS1-silencing Ad-vectored vaccines During the reporting period we: + Developed a 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. + We have tested the 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. The significance of the progress made during this period primarily relates to the 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 by attenuating the down-regulatory role of SOCS1 during the induction of immune responses will serve to significantly improve the vaccine efficacy.