New DC-targeting Vaccine Constructs and Antibody Reagents

在第一次U19中，我们建立了高特异性和高亲和力的抗人 以下人类DC受体：LOX-1，BDCA-2，DC-SIGN/L，Dectin-1，CD1d，DC-ASGPR，Langerin， CLEC-6、MARCO、CD40和DCIR。这些人类DC受体试剂发现了基本的新 并用于开发直接针对DC的抗原的原型疫苗。基于 广泛的体外药效数据，以及在两个体内模型中的初步研究，人类和 猕猴，我们假设最有效的抗原靶向疫苗将提供 抗原，同时与两个受体结合，并将直接与佐剂相连。我们设想有三个 作为目标1-3的基础的需要改进的领域。在目标1中，我们将构建多价疫苗 由流感病毒血凝素(HA)、核蛋白(NP)和基质蛋白2(M2)组成。三重 疫苗将诱导广泛的细胞和体液免疫反应。在目标2中，我们将 产生针对两个DC受体的疫苗，以增强疫苗的免疫原性。在《目标3》中， 将生产与佐剂挂钩的DC靶向疫苗。在此产生的疫苗将通过 通过AIM 4分析DC结合、激活和内化，然后在 项目1-4。项目1和项目2通过体外分析，发现最有潜力的候选者 疫苗--尤其是那些能引起粘膜免疫的疫苗。选定的疫苗将进行测试 通过项目3和项目4彻底在体内进行。

In the 1st U19, we established panels of high specificity and high affinity monoclonal antibodies against the following human DC receptors: LOX-1, BDCA-2, DC-SIGN/L, Dectin-1, CD1d, DC-ASGPR, Langerin, CLEC-6, MARCO, CD40 and DCIR. These human DC receptor reagents uncovered fundamental new biology and were used for development of prototype vaccines that target antigens directly to DCs. Based on extensive efficacy data in vitro, as well as preliminary studies in two in vivo models, the Humouse and Rhesus macaque, we hypothesize that the most potent antigen-targeting vaccines will deliver multiple antigens, engage two receptors simultaneously, and will be directly linked to adjuvants. We envision three areas for improvement that are the basis of Aims 1-3. In Aim 1, we will construct polyvalent vaccines composed of influenza viral hemagglutinin (HA), nucleoprotein (NP), and matrix protein2 (M2). Triple vaccines will induce broad repertoires of both cellular and humoral immune responses. In Aim 2, we will generate vaccines that target two DC receptors to enhance the immunogenicity of vaccines. In Aim 3, adjuvant-linked DC-targeting vaccines will be produced. The vaccines generated herein will undergo analysis of DC binding, activation, and internalization through Aim 4, and then undergo further studies within Projects 1-4. Projects 1 and 2 serve, through in vitro analysis, to discover the most potent candidate vaccines - particularly those that will evoke mucosal immunity. The selected vaccines will be tested thoroughly in vivo via Projects 3 and 4.