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

• 批准号: 10689682
• 负责人: Erik W Settles
• 金额: $ 31.55万
• 依托单位: NORTHERN ARIZONA UNIVERSITY
• 依托单位国家: 美国
• 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/10689682
• 关键词: 2019-nCoV; Acceleration; 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; HIV Infections; Human; Immune; 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; Sorting; Specimen; Symptoms; T cell receptor repertoire sequencing; T cell response; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Epitopes; Techniques; Test Result; Testing; 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

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.

项目摘要/摘要 球孢子菌病或谷热(VF)是由球孢子菌和球孢子菌引起的。 在亚利桑那州和加利福尼亚州，Posadasii的报告流行率最高。由于相似的临床症状 VF经常被误诊和误诊为细菌性或病毒性肺炎，导致 在不必要的抗生素和更长的时间来解决疾病。患者失去生产力，并 有好几个月的症状。5%的有症状病例会出现严重的CM，但可以 要有生命危险。T细胞反应在疫苗诱导的动物保护中起重要作用 模特们。事实上，这一活动似乎对人类防御球虫至关重要，因为 由于HIV感染而导致CD4+T细胞计数低的人的数量增加了，这一事实证明了这一点 出现严重室颤的风险。这项提议的目标是识别T细胞克隆及其相关的 在人类患者和动物模型中产生的表位。球孢子虫Coccidioidesspp.表位，我们 将重点放在先前确认的和新的体内早期表达的抗原上，我们将把这些 使用T细胞肽刺激策略将表位分配给他们的TCR。我们的假设是表位 从这些球虫抗原驱动患者的TCR档案，因此将包含“公共的”TCR 可用于VF诊断和确定候选疫苗优先顺序的序列。我们预测， TCR图谱可指示免疫和疾病状态。我们将通过深入探讨这一假设 球虫多肽表位刺激活化的T细胞的TCR测序。我们将确定 亚利桑那州患者样本中这些和其他球虫蛋白的TCR。 Posadasii)和加利福尼亚(C.immitis)。我们还将确认小鼠和猕猴T细胞检测到 来自这些抗原的表位帮助球虫挑战模型的发展。此外， 免疫原性球虫抗原将在这些动物模型中进行保护评估 接种疫苗。最后，我们将生成一个TCR测序诊断测试来检测VF患者。 具体地说，我们的目标是：1)鉴定弓形虫和弓形虫的表位和相关的T细胞 用T细胞受体测序和标准细胞分选法测定人标本中的克隆(TCR) 技术，2)确定在动物模型(小鼠和猪尾)中识别的TCR和表位 猕猴)挑战、自然暴露和RNA/DNA疫苗接种，以及3)比较敏感性 使用流行和非流行人群的传统血清学和TCR测序之间的差异。如果 成功后，该项目将向社区提供一套新的VF T细胞表位 人类、老鼠和猕猴。TCR序列可用于开发下一代 诊断和辅助疫苗接种的抗原识别。最后，针对这些的T细胞克隆型 表位将通过模型和VF患者的疾病结局与保护相关。