Development of a Malaria DNA Vaccine with Enchanced Immunogenicity

开发具有增强免疫原性的疟疾 DNA 疫苗

• 批准号: 7530124
• 负责人: Richard B. Markham
• 金额: $ 24.6万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-10 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7530124
• 关键词: Adjuvant; Antibodies; Antigen Presentation; Antigen-Presenting Cells; Antigens; Area; B-Lymphocyte Epitopes; Biological Assay; Biological Models; Blood; CD8B1 gene; CRISP Thesaurus; Cells; Cellular Immunity; Cessation of life; Chimeric Proteins; Class; Clinical Trials; DNA; DNA Vaccines; DNA delivery; Dendritic Cells; Development; Electroporation; Ensure; Enzyme-Linked Immunosorbent Assay; Epitopes; Erythrocytes; Evaluation; Flow Cytometry; Gene Expression; Hepatocyte; Human; Humoral Immunities; Immune; Immune response; Immune system; Immunity; Immunization; Immunofluorescence Immunologic; Inbred BALB C Mice; Infection; Interferon Type II; Intramuscular Injections; Ligands; Link; Liver; Macrophage Inflammatory Proteins; Malaria; Malaria Vaccines; Measures; Mediating; Membrane Proteins; Memory; Modeling; Molecular Profiling; Mus; Outcome; Parasitemia; Plasmids; Plasmodium yoelii; Play; Polymerase Chain Reaction; Proteins; Public Health; Recruitment Activity; Research; Ribosomal RNA; Role; Sampling; Site; Sporozoites; Staging; Standards of Weights and Measures; Stem cells; Stimulus; T memory cell; T-Lymphocyte; T-Lymphocyte Epitopes; Techniques; Testing; Time; Tissues; Tumor-Derived; Upper arm; Vaccination; Vaccines; Virus; antigen processing; base; chemokine; cytokine; design; immunogenicity; in vivo; innovation; interest; nonhuman primate; novel; novel strategies; plasmid DNA; protein expression; receptor; response; uptake; vaccination strategy; vaxfectin

DESCRIPTION (provided by applicant): Although DNA vaccines offer many advantages, their inability to elicit sustained protective immune responses in non-human primates and humans has been disappointing. Based on substantial evidence derived from tumor model systems and preliminary studies with a candidate Plasmodium yoelii vaccine, this proposal hypothesizes that an effective DNA-based vaccine against malaria can be developed by expressing malarial protein antigens as fusion proteins linked to chemokine ligands for receptors on immature dendritic cells (iDC). This vaccine model differs from related approaches in that it attracts the appropriate antigen-presenting cell to the inoculation site and by fusing the vaccine epitopes of interest to the chemokine, ensures the efficient uptake of the antigen for processing. This hypothesis will be tested in a murine model of Plasmodium yoelii infection and immunogenicity. Specifically, we will 1. Evaluate ex vivo the responses elicited in BALB/c mice by a candidate multi-epitope DNA vaccine administered by in vivo electroporation that targets the pre-erythrocytic stages of P. yoelii. The candidate vaccine to be evaluated will consist of a plasmid that expresses macrophage inflammatory protein 31 (MIP- 31) fused to (a) The SYVPSAEQI immunodominant P. yoelii Class I-restricted T cell epitope in BALB/c mice (b) The (QGPGAP)4 immunodominant P. yoelii B cell epitope in BALB/c mice and (c) The YNRNIVNRLLGDALNGKPEEKA immunodominant Class II-restricted T cell epitope in BALB/c mice. A similar DNA construct will be created that lacks the MIP-31 component and several additional positive and negative control immunization groups will also be established. Outcomes of immunization will be measured by interferon gamma (IFN-3) Elispot assays, antibody concentrations measured in ELISA and immunofluorescence assays, and by flow cytometry evaluation the development of central memory T cells. 2. Evaluate the protective efficacy of this candidate vaccine in mouse malaria challenge models. Outcomes will be followed using real-time PCR to quantitate malaria gene expression in liver samples from the mice and levels of blood parasitemia attained in immune compared to nonimmune mice. PUBLIC HEALTH RELEVANCE: This project is designed to develop a vaccine for malaria. Should this approach be successful, this vaccine could save the more than 1 million deaths that occur annually as a result of this infection.

描述（由申请人提供）：尽管DNA疫苗提供了许多优势，但它们无法引起非人类灵长类动物和人类中持续的保护性免疫反应令人失望。基于源自肿瘤模型系统的大量证据和使用候选质子质子疫苗疫苗的初步研究，该提议假设可以通过表达与非成熟齿状细胞的趋化因子蛋白相关的融合蛋白作为融合蛋白（融合蛋白）来开发针对疟疾的有效基于DNA的疫苗。该疫苗模型与相关方法不同，因为它吸引了适当的抗原细胞进入接种位点，并通过将感兴趣的疫苗表位融合到趋化因子上，从而确保有效地摄取抗原用于加工。该假设将在疟原虫感染和免疫原性的鼠模型中进行检验。具体而言，我们将进行1。在体内通过体内电穿孔施用的候选候选多角色DNA疫苗在BALB/c小鼠中产生的反应，该反应靶向p。p.yoElii的预直发阶段。 The candidate vaccine to be evaluated will consist of a plasmid that expresses macrophage inflammatory protein 31 (MIP- 31) fused to (a) The SYVPSAEQI immunodominant P. yoelii Class I-restricted T cell epitope in BALB/c mice (b) The (QGPGAP)4 immunodominant P. yoelii B cell epitope in BALB/c mice and (c) BALB/C小鼠中的Ynrnivnrllgdalngkpeeka免疫主导II限制性T细胞表位。将创建一个类似的DNA构建体，该构建体缺乏MIP-31成分，还将建立几个额外的阳性和阴性对照组。免疫的结果将通过干扰素伽马（IFN-3）ELISPOT分析，在ELISA中测得的抗体浓度和免疫荧光测定法以及流式细胞仪评估中央记忆T细胞的发展。 2。评估该候选疫苗在小鼠疟疾挑战模型中的保护性疗效。与非免疫小鼠相比，使用实时PCR遵循结局，以定量来自小鼠的肝样本中疟疾基因表达以及在免疫中获得的血液寄生虫水平。公共卫生相关性：该项目旨在开发疟疾疫苗。如果这种方法成功，则这种疫苗可以节省由于这种感染而导致每年发生的超过100万人死亡。