Project #3:Active Vaccination and Immunotherapy Against Coccidioidomycosis

项目

• 批准号: 10541243
• 负责人: CHIUNG-YU HUNG
• 金额: $ 39.58万
• 依托单位: UNIVERSITY OF TEXAS SAN ANTONIO
• 依托单位国家: 美国
• 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/10541243
• 关键词: Active Immunization; Active Immunotherapy; 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; Macrophage; 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; Recombinants; Reporting; Research; Rodent; Safety; Speed; Subunit Vaccines; System; T-Lymphocyte; T-Lymphocyte Epitopes; Transgenic Mice; Travel; Treatment Efficacy; Tularemia; United States; Vaccination; Vaccine Antigen; Vaccines; Virulent; Yeasts; adaptive immunity; attenuation; beta-Glucans; chemotherapy; clinic ready; cytokine; delivery vehicle; design; human disease; immunogenic; improved; in vivo; manufacturing scale-up; 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

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.

项目摘要 估计每年有350,000人感染球虫。有一个 未满足的需要开发更好的化学疗法和用于主动免疫和/或免疫疗法的疫苗 针对球霉菌病（CM）。我们以前已经创建了重组嵌合多肽抗原 （RCPA1）由3个先前鉴定的球虫抗原和5的最免疫原性片段组成 人类T细胞表位。将细菌表达的RCPA1加载到酵母菌 - 葡聚糖 - 吉他蛋白颗粒（GCP）中 创建GCP-RCPA1蛋白疫苗。后者刺激了强大的TH1和TH17响应并赋予 保护人类MHC II表达HLA-DR4转基因小鼠，以防止致命的肺部感染 两种球虫剂。将蛋白质疫苗推向临床准备就绪的挑战是产生 RCPA1抗原的高量和质量并验证人类对疫苗的免疫反应。在这个 项目，我们建议使用mRNA和基于矢量的交付以及 辅助系统。疫苗接收者将在体内表达coccidioes抗原 免疫系统。我们的中心假设是使用mRNA或载体的优化基于RCPA-1的疫苗 递送/辅助平台将刺激针对CM的强大Th1和Th17免疫。这两个交付 平台可以轻松缩放以生产用于临床前和临床测试的安全疫苗。 GCP-RCPA1 蛋白质疫苗将用作比较的基准。有3个具体目标。目标1：我们将设计 DNA构建体，用于编码球虫剂RCPA1抗原的mRNA的体外转录。 IVT-MRNA会 将加载到GCP中以创建GCP-RCPA1-MRNA疫苗。第二种方法将创建基于向量的 使用Francisella Novicida突变体（FN-IGLD）的疫苗。野生型FN很少引起tularemia，而F. tularensis （ft）高度毒性。 fn-igld高度衰减，它再次保护大鼠和猕猴 肺FT感染。 fn-igld还占据了混合的Th1/th17免疫力，目前正在制定 临床安全试验。我们将创建一种FN-IGLD-RCPA1疫苗。这些平台的抗原表达水平 以及它们刺激人类单毒性THP-1细胞和鼠巨噬细胞中细胞因子产生的能力 将确定。目标2：新创建的疫苗既显示出良好的抗原表达又能够 引发细胞因子产生的功效，免疫相关性和保护性将进一步评估 麦卡尼主义。我们将应用CM和人血细胞的HLA-DR4鼠模型来描述免疫 使用全球免疫基因分析方法相关和保护机制，并确认 我们的实验室中的不符合方法。 AIM3：我们将建立CM的鼠免疫疗法模型 评估最保护性疫苗的治疗功效，或与唑或新近结合使用 在项目＃1和项目2中确定了药物。