DISCOVERY AND PRECLINICAL DEVELOPMENT OF EFFICACIOUS P. vivax PRE-ERYTHROCYTIC STAGE MALARIA VACCINES

有效的红细胞前阶段疟疾疫苗的发现和临床前开发

基本信息

批准号：
10513808
负责人：
Socrates Herrera Valencia
金额：
$ 58.22万
依托单位：
CAUCASECO SCIENTIFIC RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-11-20 至 2023-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10513808
关键词：
Acceleration Affinity Chromatography Animal Model Antibodies Antibody titer measurement Antigens Aotus primate Area Attenuated B-Lymphocytes Bioinformatics Biological C57BL/6 Mouse Cells Cessation of life Clinical Complex Confocal Microscopy Country Development Eligibility Determination Enzyme-Linked Immunosorbent Assay Epitopes Erythrocytes Evaluation Experimental Animal Model Experimental Models Formulation Future Generations Glycine Goals Hepatocyte Human Immune Immune Sera Immune response Immunity Immunization Immunoglobulin G Immunologics Incidence Individual Infection Invaded Length Life Cycle Stages Malaria Malaria Vaccines Methods Molecular Conformation Monkeys Montanide ISA-51 Mus Parasites Passive Transfer of Immunity Peptide Synthesis Phase Phase I/II Clinical Trial Plasmodium Plasmodium falciparum Plasmodium vivax Plasmodium vivax vaccine Populations at Risk Process Production Prophylactic treatment Protein Microchips Proteins Public Health Radiation Recombinant Proteins Recombinants Rodent Rotation Specificity Sporozoites Standardization Sterility Structure Surface T-Lymphocyte Techniques Technology Testing Therapeutic Transgenic Organisms Vaccinated Vaccine Design Vaccinee Vaccines Variant Virus-like particle X-Ray Crystallography clinical development cost effective cytokine design disorder risk efficacy evaluation feature detection human monoclonal antibodies immunogenic immunogenicity improved in silico inhibiting antibody innovation nanoparticle novel novel vaccines passive prophylaxis preclinical development prevent protective efficacy protein aminoacid sequence protein structure rational design research clinical testing response screening self assembly synthetic peptide synthetic protein tool vaccine candidate vaccine development vaccine formulation vaccine trial volunteer

项目摘要

Abstract A significant global malaria incidence reduction in the last two decades has stimulated greater efforts toward its global eradication; however, this ambitious goal demands novel and highly efficacious control tools, including vaccines. Although the biological complexity of the Plasmodium life cycle has prevented faster progress towards the development of a highly efficacious malaria vaccine, recent technological and scientific developments could facilitate further progress. We propose to integrate established facilities, experimental models, and standardized and novel techniques to identify and characterize P. vivax (Pv) pre-erythrocytic (PE) antigens showing vaccine potential for further clinical development. Our general hypothesis is that “immunization with selected PvPE antigen constructs can induce protective immune responses in the vertebrate host.” The overall goal of this proposal is to accelerate Pv vaccine development by identifying and characterizing classical and novel vaccine candidates with confirmed protective efficacy that could be further advanced to clinical development. Specific aims are Aim 1: Characterization of novel PvPE antigens recognized preferentially by sporozoite-vaccinated and protected individuals; Aim 2: Evaluation of the immunogenicity and protective efficacy of novel PvPE protein constructs in animal models; Aim 3: Design of self-assembled protein nanoparticle (SAPN) constructs containing antigenically relevant Pv-CSP and novel PvPE selected proteins/fragments; Aim 4: Generation of anti-Pv-CSP human monoclonal antibodies (Hu-MABs) with protective efficacy to define their precise epitopes and conformations. Methods are 1) selection and expression of early and late PvPE antigens associated with sterile protection, using Pv protein microarrays. 2) In silico prediction of B, T and CTL epitopes, and synthesis of the corresponding peptide sequences. 3) Immunological characterization, i.e., humoral (ELISA Igs/isotypes) and CMI (FACS cytokines/cell profiles) responses. 4) Immunogenicity and protective efficacy analyses of selected antigen constructs/formulations in rodents, using transgenic parasites. 5) Monkey immunogenicity, ex-vivo inhibition of spz invasion (ISI) to liver cells, protective efficacy to wild-type spz, and durability of protection. 6) Development of SAPN constructs containing the Pv-CSP variants (VK210; VK247); 7) Evaluation of the nanoparticles´ immunogenicity and protective efficacy in mice and monkeys; 8) Development of Pv-CSP Hu-MABs and analyses of their protective efficacy and fine epitope specificity; 9) Antigen-antibody (Ag::Ab) interactions analyses by X-ray crystallography. The innovation of this proposal is the use of a comprehensive, rational, and rigorous Pv protein/epitope down-selection process using unique sera/cells from malaria vaccinated protected individuals, together with protein structural analyses leading to the rational design of protective nanoparticle formulations. The significance of this proposal is to provide rapid preclinical development of 1-2 highly efficacious PvPE vaccines for future clinical evaluation and Pv-HuMABs that could be administered passively for malaria prophylaxis.

抽象的在过去的二十年中，全球疟疾发生的重大减少已激发了更大的努力全球根除；但是，这个雄心勃勃的目标需要新颖和高效的控制工具，包括疫苗。尽管疟原虫生命周期的生物复杂性已阻止了更快的进展高效的疟疾疫苗的开发，最近的技术和科学发展可能促进进一步的进步。我们建议整合已建立的设施，实验模型和标准化以及识别和表征Vivax（PV）的新技术疫苗潜力以进一步临床发育。我们的总体假设是“与选定的PVPE抗原构建体可以在脊椎动物宿主中诱导受保护的免疫反应。该建议的目标是通过识别和表征古典和表征，加速光伏疫苗的开发具有确认保护有效性的新型疫苗候选物可以进一步推进临床发展。特定目的是目标1：新型PVPE抗原的表征优选地识别孢子岩疫苗接种和受保护的个体；目标2：评估免疫原性和受保护效率动物模型中新型的PVPE蛋白构建体； AIM 3：自组装蛋白纳米颗粒（SAPN）的设计包含抗原相关的PV-CSP和新型PVPE选定蛋白/片段的构建体；目标4：具有保护效率的抗PV-CSP人类单克隆抗体（HU-MAB）精确的表位和构象。方法是1）选择和表达早期和晚期PVPE抗原使用PV蛋白微阵列与无菌保护相关。 2）在B，T和CTL表位的硅预测中，和相应肽序列的合成。 3）免疫特征，即体液（ELISA） IGS/同种型）和CMI（FACS细胞因子/细胞谱）反应。 4）免疫原性和保护效率使用转基因寄生虫分析啮齿动物中选定的抗原构建体/制剂。 5）猴子免疫原性，对SPZ侵袭（ISI）对肝细胞的抑制作用，对野生型SPZ的保护效率和保护持续时间。 6）开发包含PV-CSP变体的SAPN构建体（VK210; VK247）; 7）评估小鼠和猴子中纳米颗粒的免疫原性和保护效率； 8）发展 PV-CSP HU-mab的及其受保护效率和精细表位特异性的分析； 9）抗原抗体（AG :: AB）通过X射线晶体学分析相互作用。该提议的创新是使用使用来自独特的血清/细胞的综合，有理和严格的PV蛋白/eppitope下调过程疟疾接种疫苗的受保护个体，以及蛋白质结构分析，导致理性设计受保护的纳米颗粒公式。该提议的意义是提供快速的临床前开发1-2个高效的PVPE疫苗，用于将来的临床评估和PV-HUMAB可能是用于预防疟疾的被动施用。