T cell Epitope Discovery in Sarcoidosis

结节病中 T 细胞表位的发现

• 批准号: 10297351
• 负责人: BRENT E PALMER
• 金额: $ 61.36万
• 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/10297351
• 关键词: Address; Affect; African American; Alleles; Antigens; Aspergillus nidulans; Biological Assay; Biological Markers; Biometry; Bronchoalveolar Lavage; CD4 Positive T Lymphocytes; Cells; Chronic berylliosis; Clone Cells; Complex; Data; Detection; Diagnosis; Disease; Early Diagnosis; Epitopes; Etiology; Frequencies; Funding; Genes; Granulomatous; HLA-DR3 Antigen; HLA-DRB1; Health; Histocompatibility Antigens Class II; Human; Hybridomas; Individual; Interdisciplinary Study; Interferon Type II; Interleukin-2; Knowledge; Lead; Libraries; Link; Lung; Lung diseases; 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; Syndrome; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Epitopes; Therapeutic Intervention; United States; caucasian American; cohort; combinatorial; enzyme linked immunospot assay; 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， 案件。该疾病的特征是活化的CD 4 + T细胞在肺和其他部位的积聚 疾病活动。有证据表明，这些T细胞密切参与的发病机制， 结节病在该提案的前一版本中，我们从表达HLA-DR 3的肺CD 4 + T细胞中鉴定了CD 4 + T细胞。 Löfgren综合征（LS）受试者表达相关的T细胞受体（TCR），并确定这些TCR 来自NAD依赖性蛋白脱乙酰酶（NDPD）的识别肽在常见的空气传播中表达， 霉菌种，构巢曲霉。使用HLA-DR 3-NDPD四聚体和IFN-γ ELISPOT，我们验证了这些 研究结果显示，NDPD反应性CD 4 + T细胞的数量显著增加， LS受试者;因此，我们已经确定了一个潜在的致病剂的起源LS。对瑞典语的研究 LS受试者作为当前更新的“概念验证”，其重点是识别T细胞表位 对于来源于表达HLA-DRB 1 *11：01的结节病受试者的肺的CD 4 + T细胞，HLA等位基因 与美国白人和非洲裔美国人的结节病密切相关。因此，我们假设， 表达HLA-DRB 1 *11：01的美国结节病患者肺中扩增的CD 4 + T细胞， 在对病原性结节病抗原的应答中积累，并在HLA- DRB 1 *11：01-受限时尚。该提案利用了多学科研究团队的优势 并关注美国的结节病队列。使用单细胞RT-PCR方法，Aim 1将表征 αβTCR对表达于来自美国结节病患者肺的CD 4 + T细胞上，并产生 表达疾病相关TCR的杂交瘤。第二个具体目标将确定肽， 刺激表达目标TCR的CD 4 + T细胞杂交瘤。最终的目标将使用功能分析 和HLA-DR 11-肽四聚体，以鉴定和计数肺中的抗原特异性CD 4 + T细胞。 结节病患者，并确定这些T细胞的频率是否可以作为诊断和/或治疗结节病的生物标志物。 预后因此，使用一种新颖但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，进一步推进了我们的研究。 了解这种神秘的疾病。