Project #3:Active Vaccination and Immunotherapy Against Coccidioidomycosis

项目

基本信息

批准号：
10363482
负责人：
CHIUNG-YU HUNG
金额：
$ 38.75万
依托单位：
UNIVERSITY OF TEXAS SAN ANTONIO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-01-01 至 2026-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10363482
关键词：
Active Immunization Adjuvant Antifungal Agents Antigen Presentation Antigen-Presenting Cells Antigens Area Attenuated Azoles Benchmarking Blood Cells Blood specimen Cells Chitin Chronic Clinical Clinical Trials Clinical assessments Coccidioides Coccidioidomycosis Combined Modality Therapy DNA Fever Fluconazole resistance Formulation Francisella Francisella tularensis Gene Expression Gene Expression Profiling Genes Genetic Transcription Glucans HLA-DR4 Antigen Human Immune Immune response Immune system Immunity Immunologics Immunotherapeutic agent Immunotherapy In Vitro Individual Laboratories Lung Lung infections Macaca Mediating Messenger RNA Methods Military Personnel Modeling Mus Oryctolagus cuniculus Patients Personal Satisfaction Persons Pharmaceutical Preparations Population Preclinical Testing Process Production Proteins Protocols documentation RNA vaccine Rattus Readiness Recombinants Reporting Research Rodent Safety Speed Subunit Vaccines System T-Lymphocyte T-Lymphocyte Epitopes Transgenic Mice Travel Treatment Efficacy Tularemia United States Vaccination Vaccines Virulent Yeasts adaptive immunity attenuation base beta-Glucans chemotherapy cytokine delivery vehicle design human disease immunogenic improved in vivo macrophage mouse model mutant novel novel vaccines particle polypeptide preclinical trial protective efficacy research clinical testing resistant strain response scale up therapeutic development therapy outcome vaccine candidate vaccine development vaccine formulation vector vector vaccine vector-based vaccine

项目摘要

Project Summary An estimated 350,000 people in the United States become infected with Coccidioides annually. There is an unmet need to develop better chemotherapies and a vaccine for active immunization and/or immunotherapy against coccidioidomycosis (CM). We have previously created a recombinant chimeric polypeptide antigen (rCpa1) composed of the most immunogenic fragments of 3 previously identified Coccidioides antigens and 5 human T-cell epitopes. The bacterial-expressed rCpa1 is loaded into yeast glucan-chitin particles (GCPs) to create GCP-rCpa1 protein vaccine. The latter stimulates a robust Th1 and Th17 response and confers protection for human MHC II-expressing HLA-DR4 transgenic mice against a lethal pulmonary infection with both species of Coccidioides. The challenges to advance a protein vaccine to clinical readiness are to produce high quantity and quality of the rCpa1 antigen and to validate human immune response to the vaccine. In this project, we propose to create two types of formulation methods using mRNA and vector-based delivery and adjuvant systems. The vaccine recepients will express the coccidioidal antigen in vivo to prime their own immune system. Our central hypothesis is that an optimized rCpa-1-based vaccine using an mRNA or vector delivery/adjuvant platform will stimulate robust Th1 and Th17 immunity against CM. These two delivery platforms can be easily scaled up to produce safe vaccines for preclinical and clinical testing. The GCP-rCpa1 protein vaccine will be used as a benchmark for comparison. There are 3 specific aims. Aim 1: We will design DNA constructs for in vitro transcription of mRNAs that encode Coccidioides rCpa1 antigen. IVT-mRNAs will be loaded into GCPs to create GCP-rCpa1-mRNA vaccines. The second approach will create a vector-based vaccine using a Francisella novicida mutant (Fn-iglD). Wild typle Fn rarely causes tularemia, while F. tularensis (Ft) is highly virulent. Fn-iglD is highly attenuated and it protects both rats and macaques againt a lethal pulmonary Ft infection. Fn-iglD also elictis a mixed Th1/Th17 immunity and it is currently under formulation for clinical safety trial. We will create a Fn-iglD-rCpa1 vaccine. The antigen expression levels of these platforms and their capacity to stimulate cytokine production in human monoctyic THP-1 cells and murine macrophages will be determined. Aim 2: The newly created vaccines that show both good antigen expression and capable of eliciting cytokine production will be further evaluated for their efficacy, immune correlates and protective mecahnisms. We will apply the HLA-DR4 murine model of CM and human blood cells to delineate immune correlates and protective mechanisms using a global immune gene profiling method and confirmed with immiunlogical methods in our laboratories. Aim3: We will establish a murine immunotherapy model of CM for evaluating treatment efficacy of the most protective vaccine alone or in combination with an azole or the newly identified drugs in Project #1 and Project #2.

项目概要据估计，美国每年有 35 万人感染球孢子菌。有一个开发更好的化疗方法以及用于主动免疫和/或免疫治疗的疫苗的需求尚未得到满足对抗球孢子菌病（CM）。我们之前已经创建了重组嵌合多肽抗原 (rCpa1) 由 3 个先前鉴定的球孢子菌抗原和 5 个最具免疫原性的片段组成人类 T 细胞表位。将细菌表达的 rCpa1 加载到酵母葡聚糖-几丁质颗粒 (GCP) 中创建 GCP-rCpa1 蛋白疫苗。后者刺激强烈的 Th1 和 Th17 反应并赋予保护表达人 MHC II 的 HLA-DR4 转基因小鼠免受致命性肺部感染两种球孢子菌属。将蛋白质疫苗推进临床准备的挑战是生产高数量和高质量的 rCpa1 抗原并验证人体对疫苗的免疫反应。在这个项目中，我们建议使用 mRNA 和基于载体的递送创建两种类型的制剂方法，辅助系统。疫苗接种者将在体内表达球孢子抗原以启动自己的抗原免疫系统。我们的中心假设是使用 mRNA 或载体的优化的基于 rCpa-1 的疫苗递送/佐剂平台将刺激针对 CM 的强大 Th1 和 Th17 免疫力。这两个发货平台可以轻松扩大规模，生产用于临床前和临床测试的安全疫苗。 GCP-rCpa1 蛋白质疫苗将作为比较基准。有3个具体目标。目标1：我们将设计用于体外转录编码球孢子菌 rCpa1 抗原的 mRNA 的 DNA 构建体。 IVT-mRNA 将被加载到 GCP 中以创建 GCP-rCpa1-mRNA 疫苗。第二种方法将创建一个基于向量的使用新杀弗朗西斯菌突变体 (Fn-iglD) 的疫苗。野生型 Fn 很少引起土拉菌病，而土拉菌 F. tularensis (Ft) 具有很强的毒性。 Fn-iglD 是高度减毒的，它可以保护大鼠和猕猴免受致命的伤害肺部感染。 Fn-iglD 还可引起 Th1/Th17 混合免疫，目前正在配制用于临床安全性试验。我们将研制一种 Fn-iglD-rCpa1 疫苗。这些平台的抗原表达水平及其刺激人单核 THP-1 细胞和鼠巨噬细胞产生细胞因子的能力将被确定。目标 2：新研制的疫苗既具有良好的抗原表达能力，又能够将进一步评估诱导细胞因子产生的功效、免疫相关性和保护性机制。我们将应用 CM 和人类血细胞的 HLA-DR4 小鼠模型来描绘免疫使用全局免疫基因分析方法确定相关性和保护机制，并通过我们实验室的免疫学方法。目标3：我们将建立CM的小鼠免疫治疗模型评估最具保护性的疫苗单独或与唑类或新疫苗联合使用的治疗效果确定了项目#1 和项目#2 中的药物。