Development and evaluation of a novel SOCS1-silenced HIV vaccine

新型 SOCS1 沉默 HIV 疫苗的开发和评估

• 批准号: 7635764
• 负责人: Jerry L Blackwell
• 金额: $ 22万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-11 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7635764
• 关键词: Activated Natural Killer Cell; Adenovirus Vector; Adenoviruses; Adjuvant; Animals; Antigen Presentation; Antigen-Presenting Cells; Antigens; B-Lymphocytes; Cell physiology; Cytokine Inducible SH2-Containing Protein; Dendritic Cells; Development; Equilibrium; Evaluation; Future; Goals; HIV; HIV vaccine; HIV-1; Immune; Immune response; Immunization; Infection; Maintenance; Measures; Mediating; Memory; Messenger RNA; Methods; Molecular Profiling; Phase; Phosphotransferases; Play; Proteins; Protocols documentation; Receptor Signaling; Regulation; Reverse Transcriptase Polymerase Chain Reaction; Role; STAT protein; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Small Interfering RNA; System; Testing; Therapeutic; Toll-like receptors; Vaccination; Vaccine Design; Vaccines; Viral; Viral Antigens; Virus Diseases; Western Blotting; adenovirus mediated delivery; base; cytokine; design; immunogenicity; improved; in vivo; inhibitor/antagonist; novel; prophylactic; protein function; public health relevance; response; vaccine candidate; Activated Natural Killer Cell; Adenovirus Vector; Adenoviruses; Adjuvant; Animals; Antigen Presentation; Antigen-Presenting Cells; Antigens; B-Lymphocytes; Cell physiology; Cytokine Inducible SH2-Containing Protein; Dendritic Cells; Development; Equilibrium; Evaluation; Future; Goals; HIV; HIV vaccine; HIV-1; Immune; Immune response; Immunization; Infection; Maintenance; Measures; Mediating; Memory; Messenger RNA; Methods; Molecular Profiling; Phase; Phosphotransferases; Play; Proteins; Protocols documentation; Receptor Signaling; Regulation; Reverse Transcriptase Polymerase Chain Reaction; Role; STAT protein; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Small Interfering RNA; System; Testing; Therapeutic; Toll-like receptors; Vaccination; Vaccine Design; Vaccines; Viral; Viral Antigens; Virus Diseases; Western Blotting; adenovirus mediated delivery; base; cytokine; design; immunogenicity; improved; in vivo; inhibitor/antagonist; novel; prophylactic; protein function; public health relevance; response; vaccine candidate

DESCRIPTION (provided by applicant): Efforts aimed at stimulating immune responses by modifying human immunodeficiency virus (HIV) antigens and using various delivery systems and adjuvants have so far failed to produce a HIV vaccine that protects against primary infections, highlighting the need for the continued development of alternative and effective vaccine candidates. A common feature of many of these approaches is the use of positive immunostimulation to enhance the immune response. Less often considered in the context of vaccine design however are the inhibitory signals that serve to moderate the immune response. Antigen-presenting cells, such as dendritic cells (DC), play a critical role in the establishment and maintenance of the immune responses against HIV infection; therefore DCs are key targets for vaccines that stimulate immune signaling. Our proposal is based on findings that the suppressor of cytokine signaling 1 (SOCS1) protein functions as an antigen-presentation attenuator by restricting the Janus-activated kinase-signal transducers and activators of transcription and Toll-like receptor-signaling pathways. Inhibition of SOCS1 signaling in DCs was shown to induce higher levels of both Th1- and Th2-polarizing cytokines, resulting in the enhancement of memory, antigen-specific T- and B- cell responses and activated natural killer cells. Herein we hypothesize that the immunogenicity of an adenovirus (Ad)-based HIV vaccine will be enhanced by suppressing the SOCS1 signaling pathway. To test this hypothesis, the Specific Aims of the project are to (1) inhibit DC SOCS1 signaling with Ad-mediated delivery of SOCS1 small interfering (si) RNA, (2) evaluate the effect(s) of SOCS1-silencing on DC function, and (3) determine the effect of SOCS1-silencing in Ad-targeted DCs by measuring the vaccination efficiency of HIV antigen-specific immunes responses. To accomplish Aim 1 we will first generate Ad vectors that express SOCS1 siRNA. Then, using quantitative RT-PCR and Western blotting methods, we will show that DCs infected with the siRNA-expressing Ad produce significantly reduced levels of SOCS1 mRNA and protein. In the second aim we will determine the effects of SOCS1-silencing on DC function. This will be done by flow cytometric analyses of DC phenotypic markers and cytokines expression profiles. The third phase of the project is designed to quantitatively assess the immune responses to an HIV vaccine in animals with transiently SOCS1-silenced DCs. This aim will be accomplished by measuring HIV-specific humoral and cellular-mediated responses and the ability of the vaccination protocol to mitigate an in vivo viral challenge. The negative regulation of signal transduction pathways exerted by SOCS1 is vitally important for an appropriate response to cytokine stimulation, however by transiently tipping the balance this project represents a potentially novel direction in the design of HIV vaccines. By "inhibiting the inhibitors" of antigen-presentation, such a SOCS1, this project represents a generally applicable and alternative strategy for enhancing the potency of prophylactic and therapeutic HIV vaccines. PUBLIC HEALTH RELEVANCE: Over 40 million people are currently infected with HIV and, considering the lack of a prophylactic vaccine in the foreseeable future, this number is expected to rise exponentially particularly in the underdeveloped regions of the world. The goal of this project is to evaluate whether the efficacy of an HIV vaccine can be significantly improved by modulating key effectors of the immune response. The relevance of such an improvement lies in the development and potential global impact of a vaccine that affords protection from HIV infection.

描述（由申请人提供）：旨在通过修改人类免疫缺陷病毒（HIV）抗原以及使用各种递送系统和辅助因素来刺激免疫反应的努力，迄今未能产生艾滋病毒疫苗，以防止原发性感染，从而突出了持续发展替代性疫苗和有效疫苗候选者的需求。许多这样的方法的共同特征是使用阳性免疫刺激来增强免疫反应。然而，在疫苗设计的背景下，不太经常考虑的是抑制信号，这些信号用于缓和免疫反应。抗原呈递细胞（例如树突状细胞（DC））在对HIV感染的免疫反应的建立和维持中起着至关重要的作用。因此，DC是刺激免疫信号传导的疫苗的关键靶标。我们的建议基于发现，细胞因子信号传导1（SOCS1）蛋白的抑制剂通过限制了JANUS激活的激酶信号传感器和转录和TOLL-Liel-like受体测量途径的抑制作用作为抗原表现衰减剂。显示DC中SOCS1信号传导的抑制可诱导更高水平的Th1-和Th2极化细胞因子，从而增强记忆力，抗原特异性T-和B细胞反应以及活化的天然杀伤细胞。在此，我们假设通过抑制SOCS1信号通路来增强基于腺病毒（AD）的HIV疫苗的免疫原性。 To test this hypothesis, the Specific Aims of the project are to (1) inhibit DC SOCS1 signaling with Ad-mediated delivery of SOCS1 small interfering (si) RNA, (2) evaluate the effect(s) of SOCS1-silencing on DC function, and (3) determine the effect of SOCS1-silencing in Ad-targeted DCs by measuring the vaccination efficiency of HIV antigen-specific immunes回答。为了实现目标1，我们将首先生成表达SOCS1 siRNA的AD向量。然后，使用定量的RT-PCR和Western印迹方法，我们将表明感染了表达siRNA AD的DC会产生显着降低的SOCS1 mRNA和蛋白质水平。在第二个目标中，我们将确定SOCS1沉默对直流功能的影响。这将通过对直流表型标记和细胞因子表达谱的流式细胞仪分析来完成。该项目的第三阶段旨在定量评估具有瞬时SOCS1脱毛DC的动物中对HIV疫苗的免疫反应。该目标将通过测量HIV特异性的体液和细胞介导的反应以及疫苗接种方案减轻体内病毒攻击的能力来实现。 SOCS1施加的信号转导途径的负调节对于对细胞因子刺激的适当反应至关重要，但是，通过瞬时削减平衡，该项目代表了HIV疫苗设计的潜在新颖方向。通过“抑制抗原出现的抑制剂”这样的SOCS1，该项目代表了一种普遍适用的替代策略，用于增强预防性和治疗性HIV疫苗的效力。公共卫生相关性：目前有超过4000万人感染了艾滋病毒，并且考虑到可预见的未来缺乏预防性疫苗，预计这一数字将成倍增加，尤其是在世界欠发达地区。该项目的目的是评估通过调节免疫反应的关键效应因素，是否可以显着提高HIV疫苗的功效。这种改善的相关性在于疫苗可保护艾滋病毒感染的疫苗的发展和全球影响。