A novel T7 phage display technology to detect sarcoidosis specific antigens

新型T7噬菌体展示技术检测结节病特异性抗原

基本信息

批准号：
10524024
负责人：
Lobelia Samavati
金额：
$ 40.65万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-12-15 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10524024
关键词：
Affect Amino Acid Sequence Animal Model Antibodies Antigens Autoantibodies B-Lymphocyte Epitopes Bacteriophage T7 Bacteriophages Biological Markers Biopsy Bronchoalveolar Lavage Cells Central Nervous System Clinical Complementary DNA Complex Custom Cutaneous Data Delayed Hypersensitivity Detection Development Diagnosis Diagnostic Disease Disease Marker Disease remission Embryo Enzyme-Linked Immunosorbent Assay Etiology Eye Fibroblasts Goals Granuloma Granulomatous disease Human Hypergammaglobulinemia Immune System Diseases Immune response Immunity Immunoassay In Vitro Individual Inflammatory Learning Leukocytes Libraries Lung Mediastinal lymph node group Messenger RNA Methods Micro Array Data Modeling Mononuclear Mycobacterium tuberculosis Organ Outcome Pathogenesis Pathologic Patients Peptide Synthesis Peptides Performance Peripheral Persons Phage Display Phenotype Play Population Predictive Value Progressive Disease Pulmonary Sarcoidosis Reaction Respiratory Disease Role Sample Size Sampling Sarcoidosis Sensitivity and Specificity Serology Skin Specificity Standardization System Technology Testing Tissues Training Tuberculin Test Tuberculosis Validation Whole Blood anergy antibody detection biomarker panel cDNA Library cohort design diagnostic biomarker diagnostic panel disease diagnosis gag Gene Products immune function in vitro Model insight instrument monocyte non-caseating granulomas novel novel therapeutics specific biomarkers

项目摘要

Sarcoidosis is an inflammatory disease of unknown etiology that occurs worldwide and is characterized by granuloma formation in different organs. No specific test has been developed to diagnose this disease. Confirmation of non-caseating granuloma in tissue biopsy of involved organs in the absence of other causes is the current state of the art for diagnosing sarcoidosis. We propose to test the hypothesis that overall immunity plays a prominent role in the pathogenesis of sarcoidosis, since abnormalities of the immune function and the presence of various antibodies/autoantibodies occurs in this disorder. Sarcoidosis and tuberculosis have clinical and pathological similarities. Despite isolation of various components of Mycobacterium tuberculosis (MTB) from sarcoidosis tissues, sarcoidosis subjects react to tuberculin skin test (PPD) negatively. In contrast to sarcoidosis latently infected individuals with TB (LTBI) respond to PPD with delayed type hypersensitivity reaction. Using a high throughput method, we developed a complex cDNA library derived from tissues of sarcoidosis patients. We constructed a microarray platform from this cDNA library containing large numbers of sarcoidosis clones and immunoscreened this platform with sera from patients with sarcoidosis, controls and other respiratory diseases. We identified a panel of biomarkers/classifiers with high sensitivity and specificity that can discriminate between sera of patients with sarcoidosis, healthy controls other respiratory diseases. Thus, our technology allows us to test the hypothesis that sarcoidosis is an immune disorder triggered by a group of specific antigens, which differ from LTBI antigens. Furthermore, these specific antigen peptides are capable of inducing granuloma in vitro in sarcoidosis peripheral mononuclear cells (PBMCs). To test this hypothesis, we will selectively biopan our T7 phage cDNA display library with sera of diverse sarcoidosis and LTBI subjects to select diversified clones to increase the antigen repertoire to construct a comprehensive antigen microarray platform. Second, we propose to use a large sample size from a diversified population of sarcoidosis patients and healthy controls as well as subjects with LTBI. We would like to expand test the bioreactivity of sera obtained independently from cases and controls to identify a panel of diagnostic biomarkers/classifiers, which can discriminate between sarcoidosis and healthy controls and latent TB. Furthermore, we will validate the classifiers in independent validation sets and determine whether the discovered biomarkers/classifiers can determine the clinical outcome, especially in the progressive disease course. In Aim 3, we will synthesize the antigen peptides to develop an ELISA to determine the titers of identified antigens. Finally, we test whether these clones are capable of generating granuloma in vitro using latent, healthy controls and sarcoidosis peripheral mononuclear cells. The overall goal is to define the specific antigens initiating granuloma formation in sarcoidosis and how the immunological response to antigen leads to sarcoidosis and latent TB phenotypes.

结节病是一种未知病因的炎性疾病，发生在全球范围内，其特征是不同器官的肉芽肿形成。尚未开发特定的测试来诊断这种疾病。在没有其他原因的情况下，在涉及器官的组织活检中确认非灭绝的肉芽肿是诊断结节病的现状。我们建议检验总体免疫力的假设由于免疫功能异常和在这种疾病中发生了各种抗体/自身抗体的存在。结节病和结核病患有临床和病理相似性。尽管分离了结核分枝杆菌的各种成分（MTB）从结节病组织中，结节病受试者对结核蛋白皮肤测试（PPD）有效反应。相比之下结节病潜在地感染了TB（LTBI）对PPD的反应延迟反应。使用高吞吐量方法，我们开发了一个复杂的cDNA文库，该文库衍生自结节病患者。我们从这个cDNA库中构建了一个微阵列平台，其中包含大量结节病克隆和免疫镜，并用来自结节病，对照和对照组患者的血清其他呼吸系统疾病。我们确定了具有高灵敏度和特异性的生物标志物/分类器面板可以区分结节病患者血清，健康对照其他呼吸道疾病。因此，我们的技术使我们能够检验以下假设，即结节病是一种由A触发的免疫疾病一组特定抗原，与LTBI抗原不同。此外，这些特定的抗原肽是能够在结节症外周单核细胞（PBMC）中诱导体外肉芽肿。测试这个假设，我们将选择性地biopan我们的T7噬菌体cDNA展示库，该文库具有多种结节病和 LTBI受试者选择多元化的克隆以增加抗原库，以构建全面抗原微阵列平台。其次，我们建议使用大量的样本量结节病患者和健康对照以及LTBI受试者。我们想扩展测试与病例和对照组独立于识别诊断面板的血清生物反应性可以区分结节病和健康对照和潜在结核的生物标志物/分类器。此外，我们将在独立验证集中验证分类器，并确定是否是否发现的生物标志物/分类器可以确定临床结果，尤其是在进行性疾病中课程。在AIM 3中，我们将合成抗原肽以开发ELISA以确定的滴度确定的抗原。最后，我们测试这些克隆是否能够使用潜在的，健康的对照和结节病周围的单核细胞。总体目标是定义特定抗原在结节病中引发肉芽肿的抗原以及对抗原的免疫反应如何导致结节病和潜在结核表型。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Platelets and renal failure in the SARS-CoV-2 syndrome.

SARS-CoV-2 综合征中的血小板和肾衰竭。

DOI：
10.1080/09537104.2020.1817361
发表时间：
2021-01-02
期刊：
Platelets
影响因子：
3.3
作者：
Taha M;Sano D;Hanoudi S;Esber Z;Elahi M;Gabali A;Chopra T;Draghici S;Samavati L
通讯作者：
Samavati L

Antiphospholipid antibodies in COVID-19: a meta-analysis and systematic review.