Optimizing Vaccines by Targeting Dendritic Cells with IFNa and IFNb Inducing Adjuvants

 描述（由申请人提供）：迫切需要一种安全有效的HIV-1疫苗。人们在了解对感染的免疫反应方面投入了数十年的工作，并采取了不同的方法来开发有效预防和治疗该疾病的疫苗。虽然在动物模型中有效，但没有一种在人类中有效。人类对该病毒的免疫反应是复杂的，后者表现出显着的逃避机制。然而，在一项大型临床试验中，超过30%的接种疫苗的个体受到保护，免受感染，免疫力与针对特定病毒抗原的IgG抗体（Abs）相关。随着针对HIV-1包膜糖蛋白上的保守表位的病毒中和Ab的进一步鉴定，在开发刺激人类保护性免疫应答的疫苗方面有了新的努力。树突状细胞（DC）活化的改善是开发更有效的HIV疫苗的关键，其中强的CD 4和CD 8应答是诱导抗体和细胞介导的免疫所必需的。我们以前已经表明，组成型活性IPS-1是一种有效的分子佐剂，诱导强有力的IFN-α/B反应，并激活和成熟人类和小鼠树突状细胞。使用新方法配制的组成型活性IPS-1诱导了强有力的NF-κ B应答、高水平的IFN-α/B表达、DC活化/成熟以及参与产生适应性免疫应答的关键细胞因子（包括IL-12 p70和TNF-α）的分泌。在这里，我们提出了一种靶向疫苗构建的纳米制造，最大限度地利用这种IPS 1佐剂。Parabon（r）Inemblix（tm）药物开发平台是计算机辅助设计（CAD）软件（Parabon inSemiquio（tm）Design Studio）和自组装DNA纳米制造技术的创新组合，其可用于设计和创建多功能DNA纳米结构（即，DNA折纸）。我们建议设计和制造一种DNA折纸疫苗，其中包含两种HIV-1抗原（Ags）的基因盒：HIV-1 Gag和HIV-1 gp 140 Env Foldon。为了靶向DC，疫苗构建体将用CpG ODN 2006修饰，其结合DC上的内吞介导受体DEC-205。将在几种细胞培养模型中体外评估构建体用于DC活化和转染的功效，并在鼠模型中体内评估构建体用于免疫刺激和针对牛痘-gag攻击的保护的功效。

 DESCRIPTION (provided by applicant): There is a critical need for a safe and effective vaccine against HIV-1. Decades of work have been invested in understanding the immune response to infection, and different approaches have been taken to develop vaccines that are effective in preventing and treating the disease. While effective in animal models, none have been as effective in humans. The immune response in humans to the virus is complex with the latter demonstrating remarkable evasion mechanisms. Nevertheless, in one large clinical trial, over 30% of vaccinated individuals were protected from infection and immunity correlated with IgG antibodies (Abs) against specific viral antigens. With the further identification of viral neutralizing Abs against the conserved epitopes on the HIV-1 envelope glycoproteins, there are renewed efforts in developing vaccines that stimulate a protective immune response in humans. Improvement of dendritic cell (DC) activation is critical to the development of more efficacious HIV vaccines, where strong CD4 and CD8 responses are necessary to induce both antibody and cell-mediated immunity. We have previously shown that constitutively-active IPS-1 is a potent molecular adjuvant that induces a robust IFN-a/b response, and both activates and matures human and mouse dendritic cells. Constitutively-active IPS-1, formulated using a novel approach, induced a robust NF-kB response, high levels of IFN-a/b expression, DC activation/maturation, and the secretion of key cytokines involved in generating an adaptive immune response, including IL-12p70 and TNF-a. Here we propose nano-fabrication of a targeted vaccine construct that maximally leverages this IPS1 adjuvant. The Parabon(r) Essemblix(tm) Drug Development Platform is an innovative combination of computer-aided design (CAD) software (the Parabon inSēquio(tm) Design Studio) and self-assembling DNA nano-fabrication technology that can be used to design and create multifunctional DNA nanostructures (i.e., DNA origami). We propose to design and manufacture a DNA origami vaccine that contains a gene cassette for two HIV-1 antigens (Ags): HIV-1 Gag and HIV-1 gp140 Env Foldon. To target DCs, the vaccine construct will be decorated with CpG ODN 2006, which binds to DEC-205, the endocytosis-mediating receptor on DC. The efficacy of the construct for DC activation and transfection will be evaluated in vitro in several cell culture models and in vivo in murine models for immune stimulation and protection against a vaccinia-gag challenge.