Dengue Epitope Vaccine,Tetravalent & MHCII-Targeted

登革热表位疫苗，四价

• 批准号: 7265158
• 负责人: J. Thomas August
• 金额: $ 140.14万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7265158
• 关键词: Alleles; Amino Acid Sequence; Animal Testing; Antibodies; Antibody Formation; Antigen-Presenting Cells; Antigens; Arenavirus; B-Lymphocytes; Binding; Bioinformatics; Biological; Biological Assay; Blood specimen; Brazil; CD4 Positive T Lymphocytes; CD8B1 gene; Cellular Structures; Chimera organism; Chimeric Proteins; Class; Clinical Research; Collection; Computer Analysis; Cytotoxic T-Lymphocytes; DNA Vaccines; Data; Databases; Dengue; Dengue Hemorrhagic Fever; Dengue Virus; Development; Differential Diagnosis; Disease; Effectiveness; Epitope Mapping; Epitopes; Etiology; Facility Construction Funding Category; Flavivirus; Gene Transduction Agent; Genes; Genome; Genotype; Goals; Helper-Inducer T-Lymphocyte; Histocompatibility; Human; Immune Sera; Immune response; Immunization; Infection prevention; Lymphocyte Activation; Lymphocyte antigen; Lysosomes; Medical Research; Membrane Proteins; Modeling; Mus; Nucleic Acids; Occupational activity of managing finances; Pan Genus; Patient Selection; Patients; Peptide Sequence Determination; Peptides; Pharmaceutical Preparations; Play; Polymerase Chain Reaction; Population; Population Group; Process; Proteins; Recommendation; Role; Sampling; Serotyping; Severity of illness; Signal Transduction; Singapore; Symptoms; System; T-Cell Activation; T-Lymphocyte Epitopes; Technology; Testing; Thailand; Universities; Upper arm; Vaccine Design; Vaccines; Validation; Viral Envelope Proteins; Viral Hemorrhagic Fevers; Virus; base; cohort; design; env Gene Products; genetic vaccine; neutralizing antibody; new technology; novel; pathogen; peripheral blood; programs; prototype; response; vaccine development; vaccine effectiveness; virus identification

DESCRIPTION (provided by applicant): This multi-project proposal describes several novel technologies directed at the development of a new form of epitope-based dengue genetic vaccine encoding an ensemble of selected antigen peptide units (epitopes) containing the minimum antibody and cellular antigen sequences required for an effective vaccine applicable to all serotypes (tetravalent) and to genetically diverse human populations, and lacking cytotoxic T-cell epitopes associated with hemorrhagic fever. The combination of epitope selection, targeting to the MHC II compartment, and ex vivo analysis of the immune responses with samples of a well-characterized patient cohort can be predicted to achieve a vaccine superior to those based on other strategies, and to be applicable to other pathogens. Bioinformatic computational analysis (Project 1) will be applied to identify epitope sequences selected for binding to clusters of major histocompatability class (MHC) motifs representing multiple human lymphocyte antigen (HLA) alleles (promiscuous epitopes) and effective with each of the four dengue serotypes (pan-dengue epitopes). This will include MHC II binding motifs for epitope presentation to CD4+ helper T-cells which play a critical role in the immune response system. MHC I binding motifs of CD8+ cytotoxic T-cell epitopes will also be examined both for their possible role in prevention of infection and proposed negative role in the etiology of dengue hemorrhagic fever. A human ex vivo T-cell activation assay will be used to analyze the biological correlates of the selected epitope candidates (Project 2). Analysis of human responses to epitopes is critical for vaccine development, particularly in the response to HLA-restricted T-cell epitopes. Peripheral blood samples will be obtained from a well-characterized cohort of dengue subjects (Core B). An ex vivo lymphocyte stimulation assay will identify those epitopes that stimulate the naturally induced T- and B-cells and are promiscuous to multiple HLA alleles. The results will be used to refine the bioinformatic models, for correlations to the severity of disease, and for vaccine construction. Selected epitopes will be tested initially as peptides and subsequently as nucleic acid encoded chimeric antigens (Project 4). A novel PCR assay will be used for differential diagnosis of dengue serotypes and other diseases (Project 3). The appropriate selection of patient blood samples requires rapid and accurate differential diagnosis of other diseases with similar symptoms (other flaviviruses, arenaviruses and huntaviruses). A novel multiplex format in a polymerase chain reaction (PCR) assay will provide rapid identification of viruses and additionally is designed for specific quantification of cellular replicating virus. This assay will be applied to patient selection for the Core B cohort. A tetravalent, pan-HLA, MHC II-targeted dengue DNA vaccine (Project 4) will incorporate the epitopes into an antigen chimera directed to the MHC II compartment for the required activation of CD4+ helper T-cells by targeting signals of the lysosome-associated membrane protein (LAMP) that is colocalized by MHC II and enhances all arms of the immune response CD4+, CD8+ and antibody. Epitope mapping technologies will be applied with both mouse and human antisera to identify the minimum sequences of the viral envelope proteins effective as neutralizing antibody epitopes. The vaccine constructs will be validated for human dengue virus specific T- and B-cell responses by the human ex vivo assay system (Project 2), and by mouse immunization for neutralizing antibody response (Project 4). Core A will provide central administration and financial management of the program and Core B, the peripheral blood samples of the dengue patient cohort. Additionally, the Cores will maintain a customized relational database system designed to support and streamline all aspects of the vaccine development process. As such, it will comprise integrated database "modules" for the entry, searching, tracking, and analysis of process-critical data from the early collection of blood samples from cohort patients, through the preliminary assays, and culminating in the vaccine trials of the candidate dengue epitope-defined LAMP chimera.

描述（申请人提供）：这个多项目提案描述了几种新技术，旨在开发一种新形式的基于表位的登革热基因疫苗，其编码选定抗原肽单元的集合体（表位）含有适用于所有血清型的有效疫苗所需的最低抗体和细胞抗原序列（四价）和遗传多样性人群，并且缺乏与出血热相关的细胞毒性T细胞表位。可以预测表位选择、靶向MHC II区室和用充分表征的患者群组的样品进行免疫应答的离体分析的组合，以实现上级基于其他策略的疫苗，并且适用于其他病原体。 生物信息学计算分析（项目1）将被应用于识别表位序列，这些表位序列被选择用于与代表多种人类淋巴细胞抗原（HLA）等位基因（混杂表位）的主要组织相容性类（MHC）基序簇结合，并对四种登革热血清型（泛登革热表位）中的每一种有效。这将包括用于表位呈递给在免疫应答系统中起关键作用的CD4+辅助T细胞的MHC II结合基序。还将检查CD8+细胞毒性T细胞表位的MHC I结合基序在预防感染中的可能作用和在登革出血热病因学中的拟议负面作用。 将使用人离体T细胞活化试验分析所选候选表位的生物学相关性（项目2）。分析人类对表位的反应对于疫苗开发至关重要，特别是在对HLA限制性T细胞表位的反应中。外周血样本将从充分表征的登革热受试者队列中获得（核心B）。离体淋巴细胞刺激测定将鉴定刺激天然诱导的T细胞和B细胞并且与多个HLA等位基因混杂的那些表位。研究结果将用于完善生物信息学模型，与疾病严重程度的相关性，以及疫苗的构建。所选表位将首先作为肽进行检测，随后作为核酸编码的嵌合抗原进行检测（项目4）。 一种新的PCR检测方法将用于登革热血清型和其他疾病的鉴别诊断（项目3）。患者血液样本的适当选择需要对具有类似症状的其他疾病（其他黄病毒、沙粒病毒和汉坦病毒）进行快速准确的鉴别诊断。聚合酶链反应（PCR）试验中的新型多重形式将提供病毒的快速鉴定，此外还设计用于细胞复制病毒的特异性定量。该试验将用于核心B队列的患者选择。 四价、泛HLA、MHC II靶向登革热DNA疫苗（项目4）将表位并入针对MHC II区室的抗原嵌合体中，以通过靶向溶酶体相关膜蛋白（LAMP）的信号来激活所需的CD4+辅助T细胞，所述溶酶体相关膜蛋白（LAMP）由MHC II共定位并增强免疫应答CD4+、CD8+和抗体的所有臂。表位作图技术将应用于小鼠和人抗血清，以鉴定有效作为中和抗体表位的病毒包膜蛋白的最小序列。将通过人离体测定系统（项目2）和小鼠免疫中和抗体应答（项目4）验证疫苗构建体的人登革病毒特异性T细胞和B细胞应答。 核心A将提供该计划的中央行政和财务管理，核心B将提供登革热患者队列的外周血样本。此外，核心将维持一个定制的关系数据库系统，旨在支持和简化疫苗开发过程的各个方面。因此，它将包括集成的数据库“模块”，用于输入、搜索、跟踪和分析来自队列患者的血液样品的早期收集的过程关键数据，通过初步测定，并最终在候选登革热表位定义的LAMP嵌合体的疫苗试验中。

项目成果

MULTIPRED: a computational system for prediction of promiscuous HLA binding peptides.

• DOI: 10.1093/nar/gki452
• 发表时间: 2005-07-01
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Zhang GL;Khan AM;Srinivasan KN;August JT;Brusic V
• 通讯作者: Brusic V

Alternative complement pathway deregulation is correlated with dengue severity.

• DOI: 10.1371/journal.pone.0006782
• 发表时间: 2009-08-26
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Nascimento EJ;Silva AM;Cordeiro MT;Brito CA;Gil LH;Braga-Neto U;Marques ET
• 通讯作者: Marques ET

Prediction of HLA-DQ3.2beta ligands: evidence of multiple registers in class II binding peptides.

HLA-DQ3.2beta 配体的预测：II 类结合肽中多个寄存器的证据。

• DOI: 10.1093/bioinformatics/btl071
• 发表时间: 2006
• 期刊: Bioinformatics (Oxford, England)
• 作者: Tong,JooChuan;Zhang,GuangLan;Tan,TinWee;August,JThomas;Brusic,Vladimir;Ranganathan,Shoba
• 通讯作者: Ranganathan,Shoba

Conservation and diversity of influenza A H1N1 HLA-restricted T cell epitope candidates for epitope-based vaccines.

• DOI: 10.1371/journal.pone.0008754
• 发表时间: 2010-01-18
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Tan PT;Heiny AT;Miotto O;Salmon J;Marques ET;Lemonnier F;August JT
• 通讯作者: August JT

Seroprevalence and risk factors for dengue infection in socio-economically distinct areas of Recife, Brazil.

• DOI: 10.1016/j.actatropica.2009.10.021
• 发表时间: 2010-03
• 期刊: Acta tropica
• 影响因子: 2.7
• 作者: Braga C;Luna CF;Martelli CM;de Souza WV;Cordeiro MT;Alexander N;de Albuquerque Mde F;Júnior JC;Marques ET
• 通讯作者: Marques ET