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

HIV-1 基因产品/靶向/MHC II 抗原呈现

• 批准号: 7142503
• 负责人: J. Thomas August
• 金额: $ 40.99万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7142503
• 关键词: Africa; Algorithms; Alleles; Americas; Amino Acid Sequence; Animal Model; Animals; Antibodies; Antigen Presentation; Antigen-Presenting Cells; Antigenic Diversity; Antigens; B-Lymphocytes; Binding; Biological; Biological Assay; CD4 Positive T Lymphocytes; Chimera organism; Chimeric Proteins; Class; Classification; Clinical; Clinical Trials; Collaborations; Computer Simulation; Computers; Conserved Sequence; DNA; DNA Vaccines; Dendritic Cells; Development; Elements; Epitopes; Gagging; Genetic; Genetic Variation; Genome; Genotype; Goals; Grant; HIV; HIV-1; HIV-1 vaccine; Histocompatibility; Hot Spot; Human; Immune; Immune response; Immune system; Immunity; Immunization; India; Informatics; Leukocytes; Libraries; Lymphocyte antigen; Lysosomes; Membrane Proteins; Memory; Mus; Organelles; Patients; Peptide Sequence Determination; Peptides; Population; Probability; Proteins; Protocols documentation; Research; Research Design; Research Personnel; Role; Sequence Analysis; System; T-Cell Activation; T-Lymphocyte; T-Lymphocyte Epitopes; Testing; Toxic effect; Transgenic Mice; Upper arm; Vaccine Design; Vaccines; Validation; Viral; Viral Proteins; Virus; Yellow fever virus; adeno-associated viral vector; base; design; genome sequencing; human subject; mouse model; new technology; nonhuman primate; novel; programs; protective effect; protein aminoacid sequence; research study; response; tool; vaccine delivery

The objective of this project is to develop a human HIV-1 genetic (DNA) vaccine for global application that will include the following elements of an effective vaccine: (1) HIV-1 sequences that bind to supertype human lymphocyte antigens (HLA) that represent the majority of global human populations. (2) HIV-1 sequences that represent the global population of HIV-1 genotypes and all the major clades. (3) A vaccine that stimulates all arms of the immune system and elicits immune memory to HIV challenge. The research design is based on DNA vaccine chimeras with selected HIV-1 T cell epitope sequences inserted into the luminal domain of the lysosome-associated membrane protein (LAMP) and targeted to organelles of antigen presenting cells (ARC) that contain major histocompatability class II (MHC II) proteins for presentation of antigen epitopes to CD4+ T-cells and activation of immune memory. Targeting of endogenous antigens to the MHC II compartment is known to stimulate both T- and B-cell responses and immune memory. Epitope selection is conducted by computer modeling of all HIV-1 genome sequences with algorithms that select protein sequences that are conserved in clades A to D and contain multiple, mostly overlapping, nonameric peptide sequences (hotspots) that bind to HLA supertype alleles. These supertypes represent groups of HLA alleles that have subtle differences in their binding grooves and are present in a large proportion of the human population. Validaton of T-cell activation by the selected sequences is conducted by immunizing HLA transgenic mice with DNA constructs encoding the MHC II- targeted sequences and analyzing epitope-specific T-cell responses with overlapping peptides spanning the epitope hotspot sequences. Further correlation of the selected HIV-1 sequences to human immune responses will be in collaboration with other investigators to carry out ELISpot analysis of the ex vivo T-cell responses to the selected peptide sequences of leucocytes of patients infected by HIV-1 of all major clades. Additionally, the biological role of the selected epitope sequences will be analyzed with a mouse model yellow fever virus (YFV) challenge assay system. The protective effect of immunization with the LAMP/HIV epitope chimera will be assayed by challenging the immunized animal with YFV containing the same HIV sequence. Relevance: This project proposes the application of novel technologies to the development of an effective HIV-1 vaccine applicable to global populations and all major HIV-1 clades, and capable of eliciting immune memory.

该项目的目标是开发一种供全球应用的人类HIV-1基因（DNA）疫苗， 将包括有效疫苗的以下要素：（1）结合超型人HIV-1序列 淋巴细胞抗原（HLA）代表了全球大多数人群。(2)HIV-1序列 代表了全球HIV-1基因型和所有主要分支的人群。(3)的疫苗 刺激免疫系统的所有分支，增强对HIV攻击的免疫记忆。 该研究设计基于具有选定的HIV-1 T细胞表位序列的DNA疫苗嵌合体 插入溶酶体相关膜蛋白（LAMP）的管腔结构域，并靶向 含有主要组织相容性II类（MHC II）蛋白的抗原递呈细胞（ARC）细胞器 用于将抗原表位呈递给CD 4 + T细胞和激活免疫记忆。靶向 已知MHC II区室的内源性抗原刺激T细胞和B细胞应答， 免疫记忆表位选择是通过计算机模拟所有HIV-1基因组序列进行的， 算法选择在进化枝A到D中保守的蛋白质序列，并且包含多个，主要是 重叠的九聚体肽序列（热点），其结合HLA超型等位基因。这些超类型 代表HLA等位基因组，其结合沟具有细微差异，并存在于 占人口的很大比例。通过所选序列验证T细胞活化， 通过用编码MHCII靶向的DNA构建体免疫HLA转基因小鼠进行， 序列和分析具有跨越表位的重叠肽的表位特异性T细胞应答 热点序列所选HIV-1序列与人类免疫应答的进一步相关性将在下文中描述。 与其他研究人员合作，进行体外T细胞对 感染所有主要进化枝的HIV-1的患者的白细胞的选定肽序列。另夕h 将用小鼠模型黄热病病毒分析所选表位序列的生物学作用 (YFV)激发试验系统。用LAMP/HIV表位嵌合体免疫的保护作用将 通过用含有相同HIV序列的YFV攻击免疫动物来测定。 相关性：该项目提出了新技术的应用，以开发一种有效的 适用于全球人群和所有主要HIV-1进化枝的HIV-1疫苗， 记忆