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，有效地刺激人类的所有手臂 免疫应答机制适用于人类HIV-1 DNA疫苗。这是一个新的焦点 研究的目标是将抗原作为包含溶酶体膜蛋白序列的嵌合体 (LAMP)将嵌合体运送到主要组织相容性II(MHC II)隔间以获得抗原 处理和呈现。这种方法旨在增加抗原特异性的CD4T辅助细胞(Th) 淋巴细胞反应是最佳CD8和体液免疫反应的关键。我们的研究 LAMP/GAG嵌合体疫苗表明，将抗原靶向MHC II隔室 结果小鼠各臂免疫反应、抗体、辅助性T细胞、CD4细胞均显著增加 CD8细胞毒性T细胞，CD4抗体反应增加10-100倍 由编码野生型抗原的DNA引起。我们提出的研究有以下具体目标：(1) 针对LAMP靶向HIV-I Gag的灵长类动物免疫反应的分析正在主持下进行 以确定小鼠对编码HIV-1 LAMP/Gag的DNA的增强免疫反应 嵌合体适用于非人类灵长类动物。我们将合成人类灯/Gag嵌合体，验证 抗原的正确表达和运输，并检测与之相关的免疫反应 免疫过的猴子。编码Env、Rev、Rv、 TAT和Nef作为可能的人类临床研究的其他候选者也将启动。(2)树突状细胞 特定的蛋白质，DC-LAMP和C型凝集素内吞受体，也可以运输到MHC II 隔室，并展示了不同于多细胞灯靶向的插嘴嵌合体的性质 系统，正在分析可能独特的免疫反应机制，这些机制可能对 HIV-1 DNA疫苗配方。(3)腺相关病毒载体(AAV) 我们正在用我们的DNA疫苗配方进行前景看好的基因输送工具。初期前景看好 延长免疫原性和增强对DNA初始、AAV Boost方案的抗体反应的结果， 鼓励继续调查。 Performmanceite(S)(组织、市、州) 约翰霍普金斯大学医学院，巴尔的摩，马里兰州 约翰霍普金斯新加坡生物医学中心，新加坡 关键人员。请参阅说明。根据需要使用继续页，以下面显示的格式提供所需的信息。 从首席调查员开始。按字母顺序列出所有其他关键人员，姓氏在前。 名称组织角色项目 约翰·霍普金斯大学医学院首席研究员 约翰霍普金斯大学医学院副研究员，医学博士/博士 约翰霍普金斯大学医学院博士后 李奥，伊希德，约翰霍普金斯大学医学博士 李琛，戴尔，约翰霍普金斯博士，新加坡生物博士。中心博士后 Arruda-Hinds，Luciana Johns HopkinsUniversity医学院预科博士生 约翰·霍普金斯博士，新加坡生物中心博士后 披露许可声明。适用于SBIR/STRONNY。请参阅说明。[]是[]否 小灵通398(05/01版)第2页表格第2页 在整个应用程序的底部连续编号页码。不要使用3a、3b等后缀。 H首席调查员/方案主任(最后、第一、中间)：奥古斯特，J.托马斯 必须在每张打印页和每张续页的顶部提供首席调查员/项目主任的姓名。 研究补助金 目录 页码 主页...........................................................................................................................................1 描述，

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,