New DC-targeting Vaccine Constructs and Antibody Reagents

在第一届U19中，我们建立了针对HLA-B1的高特异性和高亲和力单克隆抗体组。 下列人DC受体：LOX-1，BDCA-2，DC-SIGN/L，Dectin-1，CD 1d，DC-ASGPR，Langerin， CLEC-6、MARCO、CD 40和DCIR。这些人类DC受体试剂揭示了新的 这些基因被用于开发将抗原直接靶向DC的原型疫苗。基于 广泛的体外疗效数据，以及两种体内模型（Humouse和 恒河猴，我们假设，最有效的抗原靶向疫苗将提供多个 抗原同时与两种受体结合，并将直接与佐剂连接。我们设想三个 作为目标1-3基础的改进领域。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应答。在目标2中，我们 产生靶向两种DC受体的疫苗以增强疫苗的免疫原性。在目标3中， 将生产与免疫相关的DC靶向疫苗。在此产生的疫苗将经历 通过Aim 4分析DC结合、活化和内化，然后在 项目1-4。项目1和2通过体外分析，发现最有效的候选物 疫苗-特别是那些能激发粘膜免疫力的疫苗。将对选定的疫苗进行测试 通过项目3和项目4进行彻底体内试验。

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.