Early in vivo expressed antigens and their role in virulence, immune response, and vaccines for coccidioidomycosis

早期体内表达的抗原及其在球孢子菌病毒力、免疫反应和疫苗中的作用

基本信息

批准号：
10356629
负责人：
Erik W Settles
金额：
$ 27.34万
依托单位：
NORTHERN ARIZONA UNIVERSITY
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：
2022-08-24 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10356629
关键词：
2019-nCoV Address Animal Model Animals Antibiotics Antibody Response Antigens Arizona Attenuated Bacterial Pneumonia Biological Assay Breeding CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes COVID-19 California Cell Count Cell Separation Cells Clinical Clone Cells Coccidioides Coccidioides immitis Coccidioides posadasii Coccidioidomycosis Communities Complex County Cryopreservation DNA Diagnosis Diagnostic Diagnostic Procedure Diagnostic tests Disease Disease Outcome Epitopes Future Goals Gold HIV Infections Human Immune System Diseases Immune response Immunity Immunize Individual Infection Life Link Macaca Macaca nemestrina Medical Methods Modeling Mus Nucleic Acid Vaccines Patients Peptide Synthesis Peptides Performance Peripheral Blood Mononuclear Cell Phenotype Play Population Prevalence Primates Privatization Productivity Proteins RNA Reporting Research Project Grants Resources Risk Role Sampling Serology Serum Specimen Symptoms T cell receptor repertoire sequencing T cell response T-Cell Receptor T-Lymphocyte T-Lymphocyte Epitopes Techniques Test Result Testing Time Universities Vaccinated Vaccination Vaccines Viral Pneumonia Virulence Virulent Washington Whole Blood Work accurate diagnosis clinical research site cross reactivity desert fever disease diagnosis experience fungus human disease immunogenic immunogenicity in vivo model development novel novel diagnostics pathogen pulmonary symptom rapid diagnosis receptor response vaccine candidate vaccine development vaccine strategy vaccine-induced immunity

项目摘要

PROJECT SUMMARY/ABSTRACT Coccidioidomycosis or Valley Fever (VF), is caused by the fungus Coccidioides immitis and C. posadasii with highest reported prevalence in Arizona and California. Due to similar clinical presentations, VF is frequently misdiagnosed and mistreated as bacterial or viral pneumonia, resulting in unnecessary antibiotics and longer times to resolve illness. Patients lose productivity and experience symptoms for many months. Severe CM manifests in <5% of symptomatic cases but can be life-threatening. T cell responses plays an important role in vaccine induced protection in animal models. In fact, this activity appears to be critical for defense against Coccidioides in humans, as evidenced by the fact that humans with low CD4+ T cell counts due to HIV infection are at increased risk of severe VF. The objective of this proposal is to identify T cell clones and their associated epitopes that are generated in human patients and animal models. For Coccidioides spp. epitopes, we will focus on previously confirmed and novel in vivo early expressed antigens and we will link these epitopes to their TCRs using a T cell peptide stimulation strategy. Our hypothesis is that epitopes from these Coccidioides antigens drive patient TCR profiles and, hence, will contain “public” TCR sequences that can be used for VF diagnosis and prioritize vaccine candidates. We predict that the TCR profiles are indicative of immune and disease states. We will address this hypothesis by deep TCR sequencing of activated T cells stimulated with Coccidioides peptide epitopes. We will identify TCRs specific to these and other Coccidioides proteins in patient samples from both Arizona (C. posadasii) and California (C. immitis). We will also confirm that mouse and macaque T cells detect epitopes from these antigens to aid in Coccidioides challenge model development. In addition, the immunogenic Coccidioides antigens will be evaluated for protection in these animal models after vaccination. Finally, we will generate a TCR sequencing diagnostic test to detect VF patients. Specifically, we aim to: 1) Characterize C. posadasii and C. immitis epitopes and associated T cell clones (TCR) in human specimens using T cell receptor sequencing and standard cell sorting techniques, 2) Determine TCRs and epitopes recognized during animal model (mouse and pig-tailed macaque) challenge, natural exposure, and RNA/DNA vaccinations, and 3) Compare the sensitivity between traditional serology and TCR sequencing using endemic and non-endemic populations. If successful, this project will make available to the community a novel set of T cell epitopes for VF in humans, mice, and macaques. The TCR sequences can be used to develop next generation diagnostics and aid in antigens identification for vaccination. Finally, T cell clonotypes specific to these epitopes will be correlated to protection through the models and disease outcomes in VF patients.

项目概要/摘要球孢子菌病或谷热 (VF)，是由粗球孢子菌和粗球孢子菌引起。由于临床相似，波萨达斯在亚利桑那州和加利福尼亚州报告的患病率最高。介绍中，心室颤动经常被误诊为细菌性或病毒性肺炎，从而导致不必要的抗生素和更长的时间来解决疾病，患者会丧失生产力和。有症状的病例中出现严重 CM 的时间少于 5%，但也可能会出现症状。 T细胞反应在疫苗诱导的动物保护中发挥着重要作用。事实上，这种活性似乎对于人类防御球孢子菌至关重要，因为事实证明，由于 HIV 感染而导致 CD4+ T 细胞计数较低的人类的 CD4+ T 细胞计数增加该提案的目的是识别 T 细胞克隆及其相关的风险。对于球孢子菌表位，我们在人类患者和动物模型中产生。将重点关注先前确认的和新颖的体内早期表达抗原，我们将把这些联系起来使用 T 细胞肽刺激策略来识别 TCR 的表位。这些球孢子菌抗原驱动患者 TCR 特征，因此将包含“公共”TCR 我们预测可用于 VF 诊断和优先考虑候选疫苗的序列。 TCR 图谱表明了免疫和疾病状态，我们将深入探讨这一假设。我们将对球孢子菌肽表位刺激的活化 T 细胞进行 TCR 测序。来自亚利桑那州 (C. 我们还将确认小鼠和猕猴 T 细胞的检测。这些抗原的表位有助于球孢子菌挑战模型的开发。免疫原性球孢子菌抗原将在这些动物模型中进行保护性评估最后，我们将生成 TCR 测序诊断测试来检测 VF 患者。具体来说，我们的目标是：1) 表征 C. posadasii 和 C. immitis 表位及相关 T 细胞使用 T 细胞受体测序和标准细胞分选在人类样本中进行克隆 (TCR) 技术，2) 确定动物模型（小鼠和猪尾）中识别的 TCR 和表位猕猴）攻击、自然暴露和 RNA/DNA 疫苗接种，以及 3）比较敏感性传统血清学与使用地方性和非地方性人群的 TCR 测序之间的差异。成功后，该项目将为社区提供一组新的 VF T 细胞表位 TCR 序列可用于开发下一代。诊断并帮助疫苗接种的抗原鉴定。最后，这些是特异性的 T 细胞克隆型。表位将通过模型和 VF 患者的疾病结果与保护相关。