Dendritic Cell Targeting of Lassa Fever Vaccine

拉沙热疫苗的树突状细胞靶向

• 批准号: 6953750
• 负责人: Maria S. Salvato
• 金额: $ 22.28万
• 依托单位: UNIVERSITY OF MD BIOTECHNOLOGY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6953750
• 关键词: Lassa virus; antigen presentation; biotechnology; bioterrorism /chemical warfare; carbohydrate receptor; cell surface receptors; cellular immunity; chemical synthesis; cyclodextrins; dendritic cells; disease /disorder model; gene delivery system; gene expression; immune response; laboratory mouse; lymphocytic choriomeningitis virus; mannose; oral administration; plasmids; polyamines; reporter genes; tissue /cell culture; transfection; vaccine development; vector vaccine

DESCRIPTION (provided by applicant): Our goal is to enhance the efficacy of DNA vaccines using new bio-organic compounds to mediate delivery. Live vaccines have always been more effective than DNA vaccines in clinical trials, but recent technology allowing DNA delivery to specific cells provides new possibilities for DNA vaccines. Here we propose to test DNA vaccines for targeted delivery to dendritic cells (DC) using novel glyco-polyamines as the delivery vehicle. DC are the primary antigen-presenting cells and they express abundant cell surface carbohydrate receptors, such as mannose receptor and DC-SIGN. The cyclodextrin-based glyco-polyamines used in our studies consist of two functional domains: the multiple amino groups that bind and form complexes with DNA, and the sugar ligands that bind cell surface lectins and target the DNA complexes to DC. Enhancing the uptake of DNA by DC will enhance the immunogenicity of DNA vaccines. Cell culture studies have already demonstrated that mannosylated polyamines significantly enhance the uptake of plasmid DNA into DC. Here we propose to test the efficacy of targeted delivery in vivo. We hypothesize that DNA vaccines targeted to the cell surface carbohydrate receptors of dendritic cells will elicit enhanced cell-mediated responses. To test this hypothesis, we will initially employ the LCMV-infected murine model system that is known to elicit strong cell-mediated immune responses. We will have two specific aims. In aim 1, we will synthesize novel glycosylated cyclodextrin-based polyamines (CDPA) and test the uptake and expression of reporter genes in dendritic cell culture. Mannose and oligomannose ligands will be introduced into CDPA for targeting to mannose receptors and DC-SlGN on dendritic cells, respectively. In aim 2, we will test the magnitude of cell mediated immunity after oral inoculation with DNA vaccines in various formulations. Plasmid DNA encoding the LCMV GP gene will be orally delivered to mice as naked DNA or as DNA complexed with CDPA or glycosylated CDPA. Once we find a formulation that is optimum for eliciting cell-mediated immunity, we will replace the LCMV-GP gene with DNA encoding antigens associated with protective immunity in Lassa fever (Lassa NP and GP genes). We will determine, in the murine system, whether DNA complexed with our compound can still elicit strong cell-mediated immune responses. Once we have optimized the vaccine formulation in the murine model system, we will apply the targeted delivery system to monkey models and human clinic trials. These studies will lead to improved vaccines against Lassa fever and new compounds that will be useful for cell-specific delivery.

描述（由申请人提供）：我们的目标是使用新的生物有机化合物介导递送来增强DNA疫苗的功效。在临床试验中，实时疫苗一直比DNA疫苗更有效，但是最近允许DNA向特定细胞递送的技术为DNA疫苗提供了新的可能性。在这里，我们建议使用新型的糖过程作为输送载体测试DNA疫苗，以靶向递送向树突细胞（DC）。 DC是主要的抗原呈递细胞，它们表达丰富的细胞表面碳水化合物受体，例如甘露糖受体和DC-SIGN。我们的研究中使用的基于环糊精的糖聚酰胺由两个功能结构域组成：与DNA结合并形成复合物的多个氨基组，以及结合细胞表面凝集素并将DNA复合物靶向DC的糖配体。通过DC增强DNA的摄取将增强DNA疫苗的免疫原性。细胞培养研究已经表明，甘露糖基化的多胺显着增强了质粒DNA对DC的摄取。在这里，我们建议在体内测试目标递送的功效。我们假设针对树突状细胞细胞表面碳水化合物受体的DNA疫苗将引起增强的细胞介导的反应。为了检验该假设，我们最初将采用已知会引起强细胞介导的免疫反应的LCMV感染的鼠模型系统。我们将有两个具体的目标。在AIM 1中，我们将合成基于新型的糖基化环糊精聚胺（CDPA），并测试树突状细胞培养中报告基因的摄取和表达。将分别将甘露糖和少突糖配体引入CDPA中，以分别靶向甘露糖受体和树突状细胞上的DC-SLGN。在AIM 2中，我们将在各种制剂中用DNA疫苗进行口服接种后，测试细胞介导的免疫力的大小。编码LCMV GP基因的质粒DNA将以裸DNA或与CDPA或糖基化CDPA复合的DNA口服。一旦我们找到了一种最适合引发细胞介导的免疫力的公式，我们将用与LASSA热的保护性免疫相关的DNA替代LCMV-GP基因（LASSA NP和GP基因）。在鼠系统中，我们将确定与我们的化合物复合的DNA是否仍然可以引起强大的细胞介导的免疫反应。 一旦我们优化了鼠模型系统中的疫苗配方，我们将应用目标输送系统和人类临床试验。这些研究将导致针对LASSA发烧的疫苗和新化合物的疫苗，这些疫苗可用于细胞特异性递送。