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艾滋病毒-1疫苗。人们投入了数十年的工作来了解对感染的免疫反应，并采取了不同的方法来开发有效预防和治疗这种疾病的疫苗。虽然在动物模型上有效，但没有一种在人类上有效。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%的接种者被保护免受感染，免疫与针对特定病毒抗原的抗体(抗体)相关。随着针对HIV-1包膜糖蛋白保守表位的病毒中和抗体的进一步鉴定，人们在开发刺激人类保护性免疫反应的疫苗方面做出了新的努力。提高树突状细胞(DC)的活性是开发更有效的HIV疫苗的关键，在HIV疫苗中，强大的CD4和CD8反应是诱导抗体和细胞免疫所必需的。我们之前已经证明，具有成分活性的IPS-1是一种有效的分子佐剂，可以诱导强大的干扰素-a/b反应，并激活和成熟人和小鼠树突状细胞。使用新方法构建的结构性活性IPS-1诱导了强大的核因子-kB反应，高水平的干扰素-a/b表达，DC的激活/成熟，以及参与产生适应性免疫反应的关键细胞因子的分泌，包括IL-12p70和TNF-a。在这里，我们建议纳米制造一种靶向疫苗结构，最大限度地利用这种IPS1佐剂。Parabon(R)EsSemblix(Tm)药物开发平台是计算机辅助设计(ē)软件(Parabon INS DNAQuo(Tm)设计工作室)和自组装DNA纳米制造技术的创新结合，可用于设计和创造多功能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，该疫苗包含两个HIV-1抗原(AGS)的基因盒：HIV-1 Gag和HIV-1 gp140 Env Foldon。为了针对DC，疫苗结构将被CpG ODN 2006修饰，它与DC上的内吞介导受体DEC-205结合。将在几种细胞培养模型和体内免疫刺激和保护小鼠模型中评估用于DC激活和转染的构建物的有效性。

 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.