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: 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
• 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

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.

项目总结/摘要 球孢子菌病或山谷热（VF），是由真菌粗球孢子菌和C。 在亚利桑那州和加州报告的患病率最高。由于类似的临床 在临床表现中，VF经常被误诊为细菌性或病毒性肺炎， 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， 症状持续数月。重度CM在<5%的有症状病例中表现出来，但可以 会危及生命T细胞反应在疫苗诱导的动物免疫保护中起着重要作用 模型事实上，这种活性似乎对人类抵抗球孢子菌至关重要， 事实证明，由于HIV感染导致的CD 4 + T细胞计数低的人， 严重VF的风险。本提案的目的是鉴定T细胞克隆及其相关的免疫学特性。 在人类患者和动物模型中产生的表位。对于球孢子菌属表位，我们 将集中于先前确认的和新的体内早期表达的抗原，我们将把这些 使用T细胞肽刺激策略将表位结合到其TCR。我们的假设是， 从这些球孢子菌抗原驱动患者TCR谱，因此将包含“公共”TCR 可以用于VF诊断和优先考虑候选疫苗的序列。我们预测 TCR谱指示免疫和疾病状态。我们将通过深入讨论这一假设 用球孢子菌肽表位刺激的活化T细胞的TCR测序。我们将确定 在来自亚利桑那州的患者样本中，对这些和其他球孢子菌蛋白具有特异性的TCR（C. posadasii）和加州（C. immitis）。我们还将证实小鼠和猕猴的T细胞检测到 这些抗原的表位，以帮助球孢子菌攻击模型的发展。此外该 免疫原性球孢子菌抗原将在这些动物模型中评价保护作用， 预防针最后，我们将生成TCR测序诊断测试来检测VF患者。 具体而言，我们的目标是：1）表征C。posadasii和C.免疫表位和相关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检测TCR 2）确定在动物模型（小鼠和猪尾鼠）期间识别的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 传统血清学和TCR测序之间的差异。如果 如果成功，该项目将为社区提供一组新的VF T细胞表位， 人类、小鼠和猕猴。TCR序列可用于开发下一代 诊断和辅助抗原鉴定用于疫苗接种。最后，T细胞克隆型特异于这些 表位将通过VF患者的模型和疾病结果与保护相关。