HIV-1 Gene Products/Targeted/MHC II Antigen Presentation

描述（由申请人提供）：该项目的目标是开发一种DNA疫苗配方，有效刺激免疫反应机制的所有分支，并适用于人类HIV-1 DNA疫苗。本研究的新焦点是将抗原作为含有溶酶体膜蛋白（LAMP）序列的嵌合体靶向，该嵌合体将嵌合体运输到主要组织相容性类II （MHC II）室进行抗原加工和递呈。这种方法旨在增加抗原特异性CD4+ t辅助（Th）淋巴细胞反应，这对于最佳的CD8+和体液免疫反应至关重要。我们对LAMP/Gag嵌合体疫苗的研究表明，将抗原靶向MHC II隔室导致小鼠所有臂的免疫反应，抗体，CD4+辅助性T细胞和CD8+细胞毒性T细胞的显著增加，CD4抗体反应的增加比DNA编码野生型抗原引起的应答增加10- 100倍。我们提出的研究有以下具体目的：(1)在NIAID的支持下进行灵长类动物对LAMP靶向HIV-I Gag的免疫反应分析，以确定小鼠对编码HIV-1 LAMP/Gag嵌合体的DNA的增强免疫反应是否适用于非人灵长类动物。我们将合成人类LAMP/Gag嵌合体，验证抗原的正确表达和运输，并检测免疫猴子的相关免疫反应。将开始合成和分析编码Env、Rev、Tat和Nef的MHC II靶向DNA疫苗，作为可能的人类临床研究的其他候选疫苗。(2)树突状细胞特异性蛋白DC-LAMP和c型凝集素内吞受体，也可运输到MHC II室，并表现出与多细胞LAMP靶向系统不同的Gag嵌合体的特性，正在分析可能独特的免疫反应机制，可能证明对HIV-1 DNA疫苗配方有价值。(3)腺相关病毒载体（AAV）是最有前途的基因传递载体之一，目前正在用我们的DNA疫苗配方进行评估。延长免疫原性和增强对DNA引物的抗体反应的初步有希望的结果，AAV增强方案，鼓励继续研究。

DESCRIPTION (provided by applicant):The goals of this project are to develop a DNA vaccine formulation that effectively stimulates all arms of the immune response mechanism and is applicable to a human HIV-1 DNA vaccine. The novel focus of this research is to target the antigen as a chimera containing sequences of the lysosome membrane protein (LAMP) that traffic the chimera to the major histocompatability class II (MHC II) compartment for antigen processing and presentation. This approach is designed to increase antigen-specific CD4+ T-helper (Th) lymphocyte responses that are critical for optimal CD8+and humoral immune responses. Our research with the LAMP/Gag chimera vaccine has shown that targeting of the antigen to the MHC II compartment results in a marked increase in all arms of the immune responses of mice, antibody, CD4+helper T cells, and CD8+cytotoxic T cells, with increases in CD4 antibody responses 10- to 100-fold greater than those resulting from DNA encoding wild type antigen. Our proposed research has the following specific aims: (1) Analysis of primate immune responses to LAMP-targeted HIV-I Gag is being conducted under the auspices of the NIAID to determine if the enhanced immune responses of mice to DNA encoding an HIV-1 LAMP/Gag chimera is applicable to a non-human primate. We will synthesize the human LAMP/Gag chimera, validate the correct expression and trafficking of the antigen, and assay relevant immune responses of the immunized monkeys. The synthesis and analysis of MHC II targeting of DNA vaccines encoding Env, Rev, Tat, and Nef as other candidates for possible human clinical studies will also be initiated. (2) Dendritic cell specific proteins, DC-LAMP and C-type lectin endocytic receptors, that also traffic to the MHC II compartment and demonstrate properties as Gag chimeras that differ from the multicellular LAMP targeting system, are being analyzed for possibly unique immune response mechanisms that may prove of value to a HIV-1 DNA vaccine formulation. (3) Evaluation of adeno-associated virus vector (AAV), one of the most promising gene delivery vehicles, is being conducted with our DNA vaccine formulations. Initial promising results of prolonged immunogenicity and enhanced antibody response to a DNA prime, AAV boost regimen, encourage continued investigation.