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

这个项目的目标是开发一种DNA疫苗配方，有效地刺激所有的手臂， 免疫应答机制，适用于人HIV-1 DNA疫苗。这本小说的重点是 研究是将抗原作为含有溶酶体膜蛋白序列的嵌合体 （LAMP），其将嵌合体运输到主要组织相容性II类（MHC II）区室用于抗原 加工和展示。这种方法旨在增加抗原特异性CD 4+辅助性T细胞（Th） 淋巴细胞应答，这对于最佳的CD 8+和体液免疫应答至关重要。我们的研究 与LAMP/Gag嵌合体疫苗的结合表明，抗原靶向MHC II区室 导致小鼠、抗体、CD 4+辅助T细胞的免疫应答的所有臂的显著增加，以及 CD 8+细胞毒性T细胞，其中CD 4抗体应答的增加比CD 8+细胞毒性T细胞高10至100倍。 由编码野生型抗原的DNA产生。本研究的具体目标如下：（1） 灵长类动物对靶向LAMP的HIV-I Gag的免疫应答分析正在 以确定小鼠对编码HIV-1 LAMP/Gag的DNA的增强的免疫应答是否 嵌合体适用于非人灵长类动物。我们将合成人LAMP/Gag嵌合体，验证 抗原的正确表达和运输，并测定相关的免疫应答。 免疫猴子编码Env，Rev， 达特和Nef作为可能的人类临床研究的其他候选物也将启动。(2)树突状细胞 特异性蛋白质，DC-LAMP和C型凝集素内吞受体，也运输到MHC II 隔室，并证明与多细胞LAMP靶向不同的Gag嵌合体的性质 系统，正在分析可能独特的免疫反应机制，可能证明有价值的， HIV-1 DNA疫苗制剂。(3)腺相关病毒载体（AAV）是目前研究最多的病毒载体之一， 有前途的基因传递载体，正在进行与我们的DNA疫苗制剂。初步有希望 结果是延长了免疫原性和增强了对DNA初免、AAV加强方案的抗体应答， 鼓励继续调查。 地点（组织、城市、州） 约翰·霍普金斯大学医学院，巴尔的摩，马里兰州 Johns Hopkins新加坡生物医学中心，新加坡 关键人员。参见说明。根据需要使用续页，以下列格式提供所需信息。 从主要研究者开始，按顺序列出所有其他关键人员，姓在前。 名称组织机构项目 August，J.托马斯，医学博士约翰霍普金斯大学医学院首席研究员 Marques，Ernesto，MD/PhD约翰霍普金斯大学医学院研究助理 Chikhlikar，Priya，PhD约翰霍普金斯大学医学院博士后研究员 Leao，Ihid，MD约翰霍普金斯大学医学院博士后研究员 沈力陈，德尔，博士Johns Hopkins新加坡生物医学中心博士后研究员 Arruda-Hinds，Luciana Johns Hopkins霍普金斯大学医学院预科生 Maciel，MiltonJunior，PhD Johns Hopkins新加坡生物医学中心博士后研究员 披露许可声明。仅适用于SBIR/STTR。参见说明。[]是[]否 + PHS 398（修订版05/01）第2页表格第2页 +在整个应用程序的底部连续编号页面。不要使用后缀，如3a，3b。 h主要研究者/项目负责人（上、中、前）：August，J.托马斯 主要研究者/项目负责人的姓名必须在打印页和续页的顶部提供。 研究资助 目录 页码 首页......................................................................................................................................................... 1 说明，

The goals of this project are to develop a DNA vaccine formulationthat effectivelystimulates all arms of the immune response mechanismand is applicableto 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 traffick the chimera to the major histocompatability class II (MHC II) compartment for antigen processing and presention. This approach is designed to increase antigen-specific CD4+ T-helper (Th) lymphocyte responses which 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 traffick 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. PERFORMANCESITE(S) (organization,city, state) The Johns Hopkins UniversitySchoolof Medicine,Baltimore,Maryland Johns HopkinsSingaporeBiomedicalCentre, Singapore KEY PERSONNEL. See instructions. Use continuationpages as needed to providethe requiredinformationinthe format shownbelow. Startwith Principal Investigator.List all other key personnelin alphabeticalorder, last namefirst. Name Organization Roleon Project August, J. Thomas,MD Johns HopkinsUniv.Schoolof Medicine Principal Investigator Marques,Ernesto,MD/PhD Johns HopkinsUniv.Schoolof Medicine ResearchAssociate Chikhlikar,Priya, PhD Johns HopkinsUniv.Schoolof Medicine PostdoctoralFellow Leao, Ihid,MD Johns HopkinsUniv.Schoolof Medicine PostdoctoralFellow Sim Li Chen, Del, PhD Johns HopkinsSingaporeBiomed.Centre PostdoctoralFellow Arruda-Hinds,Luciana Johns HopkinsUniv.Schoolof Medicine Predoctoralstudent Maciel,MiltonJunior, PhD Johns HopkinsSingaporeBiomed.Centre PostdoctoralFellow Disclosure Permission Statement. Applicableto SBIR/STTROnly. See instructions.[] Yes [] No + PHS 398 (Rev. 05/01) Page 2 Form Page 2 +Number pages consecutively at the bottom throughout the application. Do not use suffixes such as 3a, 3b. h Principal Investigator/ProgramDirector(Last,first, middle): August, J. Thomas The nameof the principalinvestigator/programdirector must beprovided at the top of each printedpage andeach continuationpage. RESEARCH GRANT TABLE OF CONTENTS Page Numbers Face Page ......................................................................................................................................................... 1 Description,