T cell Epitope Discovery in Sarcoidosis

结节病中 T 细胞表位的发现

• 批准号: 10633277
• 负责人: BRENT E PALMER
• 金额: $ 56.61万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-05 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10633277
• 关键词: Affect; African American population; Alleles; Antigens; Aspergillus nidulans; Biological Assay; Biological Markers; Biometry; Bronchoalveolar Lavage; CD4 Positive T Lymphocytes; Cell Separation; Cells; Chronic berylliosis; Clone Cells; Complex; Data; Detection; Diagnosis; Disease; Early Diagnosis; Epitopes; Etiology; Frequencies; Funding; Genes; Granulomatous disease; HLA-DR3 Antigen; HLA-DRB1; Health; Histocompatibility Antigens Class II; Human; Hybridomas; Individual; Interdisciplinary Study; Interferon Type II; Interleukin-2; Knowledge; Libraries; Link; Lofgren Syndrome; Lung; Molds; Mus; Onset of illness; Pathogenesis; Patients; Peptides; Population; Prognosis; Protein Databases; Proteins; Pulmonary Sarcoidosis; Reverse Transcriptase Polymerase Chain Reaction; Role; Sarcoidosis; Scanning; Severity of illness; Sir2-like Deacetylases; Site; Specificity; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Epitopes; Therapeutic Intervention; United States; caucasian American; cohort; combinatorial; enzyme linked immunospot assay; granulomatous lung disease; innovation; interest; novel; novel strategies; response; screening

Project Summary Sarcoidosis is a systemic granulomatous disorder of unknown etiology that affects the lung in greater than 90% of cases. The disease is characterized by the accumulation of activated CD4+ T cells in the lung and other sites of disease activity. Evidence suggests that these T cells are intimately involved in the pathogenesis of sarcoidosis. In the previous version of this proposal, we identified lung CD4+ T cells from HLA-DR3-expressing Löfgren’s syndrome (LS) subjects expressing related T cell receptors (TCRs) and determined that these TCRs recognized peptides derived from NAD-dependent protein deacetylase (NDPD) expressed in a common airborne mold species, Aspergillus nidulans. Using HLA-DR3-NDPD tetramers and IFN-γ ELISPOT, we validated those findings and showed that a significantly greater number of NDPD-responsive CD4+ T cells exists in the lungs of LS subjects; thus,we have identified a potential causative agent in the genesis of LS. This study of Swedish subjects with LS serves as a “proof of concept” for the current renewal whose focus is to identify T cell epitopes for CD4+ T cells derived from the lungs of sarcoidosis subjects expressing HLA-DRB1*11:01, the HLA allele strongly linked to sarcoidosis in Caucasians and African-Americans in the US. Thus, we hypothesize that expanded CD4+ T cells in the lungs of HLA-DRB1*11:01-expressing US sarcoidosis patients are accumulating in response to etiologic sarcoidosis antigen(s) and recognize those antigens in an HLA- DRB1*11:01-restricted fashion. This proposal harnesses the strengths of a multidisciplinary research team and focuses on a sarcoidosis cohort in the US. Using a single cell RT-PCR approach, Aim 1 will characterize αβTCR pairs expressed on CD4+ T cells derived from the lungs of US sarcoidosis patients and generate hybridomas expressing disease-relevant TCRs. The second specific aim will determine the peptides that stimulate the CD4+ T cell hybridomas expressing the TCRs of interest. The final aim will use functional assays and HLA-DR11-peptide tetramers to identify and enumerate antigen-specific CD4+ T cells in the lungs of sarcoidosis patients and determine if the frequency of these T cells can serve as a biomarker for diagnosis and/or prognosis. Thus, using a novel yet proven scientific approach, we will address critical knowledge gaps in the etiologic T cell antigens involved in the pathogenesis of sarcoidosis in US patients, further advancing our understanding of this enigmatic disease.

项目摘要 结节病是一种病因不明的全身性肉芽肿性疾病，90%以上影响肺部。 案件的数量。这种疾病的特点是活化的CD4+T细胞在肺和其他部位积聚 疾病的活跃度。有证据表明，这些T细胞与糖尿病的发病密切相关。 结节病。在这项提议的前一个版本中，我们从表达HLA-DR3的肺组织中鉴定出了肺中的CD4+T细胞 L综合征(LS)患者表达相关T细胞受体(TCR)的研究 NAD依赖蛋白脱乙酰酶(NDPD)在常见空气中表达的识别多肽 霉菌种类，尼杜拉曲霉。用人类白细胞抗原-DR3-NDPD四聚体和干扰素-γELISPOT验证 研究结果表明，在小鼠的肺中存在明显更多的NDPD反应的CD4+T细胞 因此，我们已经确定了LS的发生中的一个潜在的致病因素。关于瑞典语的研究 患有LS的受试者为当前的更新提供了概念证明，其重点是识别T细胞表位 对于表达HLA-DRB1*11：01的结节病患者肺来源的CD4+T细胞，HLA等位基因 与美国高加索人和非裔美国人的结节病密切相关。因此，我们假设 表达HLA-DRB1*11：01的美国结节病患者肺内扩增的CD4+T细胞 对病原性结节病抗原(S)的反应并识别这些抗原 DRB1*11：01-受限时尚。这项建议利用了一个多学科研究团队的优势。 并聚焦于美国的一个结节病队列。使用单细胞RT-PCR方法，Aim 1将表征 美国结节病患者肺来源αβ+T细胞上表达的CD4TCR对 表达疾病相关TCRs的杂交瘤。第二个特定目标将确定 刺激表达TCRs的CD4+T细胞杂交瘤细胞。最终目标将使用功能分析。 和人类白细胞抗原-DR11多肽四聚体鉴定和计数小鼠肺组织中抗原特异性的CD4+T细胞 并确定这些T细胞的频率是否可以作为诊断和/或 预后。因此，使用一种新颖而又经过验证的科学方法，我们将解决 病原性T细胞抗原参与了美国患者结节病的发病机制，进一步推动了我们的 对这种神秘疾病的了解。

项目成果

CD4+ T cells in the lungs of acute sarcoidosis patients recognize an Aspergillus nidulans epitope.

• DOI: 10.1084/jem.20210785
• 发表时间: 2021-10-04
• 期刊: The Journal of experimental medicine
• 作者: Greaves SA;Ravindran A;Santos RG;Chen L;Falta MT;Wang Y;Mitchell AM;Atif SM;Mack DG;Tinega AN;Maier LA;Dai S;Pinilla C;Grunewald J;Fontenot AP
• 通讯作者: Fontenot AP

Innate and Adaptive Immunity in Noninfectious Granulomatous Lung Disease.

• DOI: 10.4049/jimmunol.2101159
• 发表时间: 2022-04-15
• 期刊: Journal of immunology (Baltimore, Md. : 1950)

Shared αβ TCR Usage in Lungs of Sarcoidosis Patients with Löfgren's Syndrome.

• DOI: 10.4049/jimmunol.1700570
• 发表时间: 2017-10-01
• 期刊: Journal of immunology (Baltimore, Md. : 1950)
• 作者: Mitchell AM;Kaiser Y;Falta MT;Munson DJ;Landry LG;Eklund A;Nakayama M;Slansky JE;Grunewald J;Fontenot AP
• 通讯作者: Fontenot AP