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。为了验证这一假设，该项目的具体目标是： 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)评价SOCS1沉默Ad载体疫苗诱导的HIV特异性免疫应答 在这一筹资期间，我们取得了以下进展： +开发了共表达靶向小鼠SOCS1 mRNA和转基因的siRNA的腺病毒载体 +证实SOCS1 siRNA沉默降低了小鼠骨髓来源的小鼠树突状细胞中SOCS1蛋白的表达 +证明SOCS1 siRNA沉默延长了由脂多糖和干扰素-γ信号诱导的小鼠骨髓来源树突状细胞中STAT1磷酸化的持续时间 +SOCS1沉默型腺病毒载体的初步试验显示，体内抗原特异性CD8+T细胞反应增强 在接种SOCS1沉默载体的小鼠中观察到更高水平的腺病毒特异性CD8+T细胞 +产生了三个腺病毒载体，表达针对猕猴SOCS1 mRNA的SOCS1 siRNA 我们的进展的意义在于开发了一种新型的腺病毒载体平台，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. To test this hypothesis, the Specific Aims of the project are 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 We have made the following progress during this funding period: + Developed adenovirus vectors that co-express siRNA that target mouse SOCS1 mRNA and transgenes + Demonstrated the SOCS1 siRNA silencing reduced the expression of SOCS1 protein levels in mouse bone marrow-derived mouse dendritic cells + Demonstrated that SOCS1 siRNA-silencing extends the duration of STAT1 phosphorylation as a result of induction by LPS and INF-gamma signaling in bone marrow-derived mouse dendritic cells + Pilot test of the SOCS1-silencing adenovirus vectors demonstrated enhanced antigen-specific CD8+ T cells responses in vivo + Observed higher levels of adenovirus-specific CD8+ T cells in mice vaccinated with SOCS1-silencing vectors + Generated three adenovirus vectors that express SOCS1 siRNAs that target rhesus macaque SOCS1 mRNA The significance of our progress is in the development of a novel adenovirus vector platform 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.