Identifying Specific Antigenic Targets of Kawasaki Disease

识别川崎病的特定抗原靶点

基本信息

批准号：
10686007
负责人：
ANNE H ROWLEY
金额：
$ 42.73万
依托单位：
LURIE CHILDREN'S HOSPITAL OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-22 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10686007
关键词：
Amino Acid Substitution Aneurysm Antibodies Antibody Response Antigen Targeting Antigens Autoantigens Binding Biological Assay Cells Cessation of life Child Childhood Clinical Clonal Expansion Coronary artery Data Developed Countries Development Diagnosis Diagnostic tests Disease Enzyme-Linked Immunosorbent Assay Epidemiology Epitopes Etiology Fever Future Geographic Locations Goals Health Care Costs Heart Diseases High-Throughput Nucleotide Sequencing Human Immune Targeting Immune response Immunoglobulin A Immunoglobulin G Immunoglobulin M Immunoglobulins Inclusion Bodies Infant Infection Investigation Libraries Mass Spectrum Analysis Messenger RNA Monoclonal Antibodies Mucocutaneous Lymph Node Syndrome Myocardial Infarction Nucleic Acids Pathogenesis Patients Pediatrics Peptides Phage Display Plasmablast Predisposition Prevention Protein Array Proteins RNA RNA Sequences Reverse Transcriptase Polymerase Chain Reaction Risk Serology Serology test Sorting Testing Time Tissues Viral Virus Western Blotting bronchial epithelium cDNA Library detection assay diagnostic assay immunogenic improved laser capture microdissection novel therapeutics peripheral blood protein purification response screening viral detection

项目摘要

PROJECT SUMMARY/ABSTRACT Kawasaki Disease (KD) is the leading cause of acquired heart disease in children in developed nations. KD can result in coronary artery aneurysms that can lead to lifelong heart disease, myocardial infarction, and death. The clinical and epidemiologic features support an infectious etiology in genetically susceptible children, but the cause has eluded more than 50 years of study. Delayed and missed diagnoses increase the risk of coronary artery aneurysms. The development of urgently needed diagnostic tests and improved therapies are dependent upon identifying the etiology. In preliminary studies, we analyzed the peripheral blood plasmablast response at 1-3 weeks after KD fever onset using single cell RT-PCR and made 60 monoclonal antibodies (Mab) from these plasmablasts. We used these Mab to determine their target antigens. We found that 32/60 Mab, derived from 9/11 KD patients, identify intracytoplasmic virus-like inclusion bodies (ICI) in ciliated bronchial epithelium of KD children but not infant controls. Using peptide arrays to identify binding epitope(s), amino acid substitution arrays, and ELISA, we found that 5 of the 32 (16%) Mab, derived from 3 KD patients with coronary artery aneurysms, recognize KD peptide epitope 1. KD peptide 1 completely blocks binding of these Mab to KD ICI, indicating that a protein with the binding epitope sequence is present in the ICI. Moreover, pre-treatment sera from 5/8 KD patients >day 8 after fever onset but 0/17 infant control sera have IgG antibody to the peptide epitope. We recently identified two additional epitopes using this approach that require further study. These findings support our hypothesis that at least a subset of KD cases results from infection with a presently unidentified, ubiquitous viral causative agent containing the KD peptide epitope 1 sequence. In this proposal, we hypothesize that identification of antigens targeted by the plasmablast response to KD can lead to identification of the etiology(ies) and development of KD serologic tests. To test these hypotheses, we will: 1) identify the specific proteins recognized by our panel of KD Mab that bind to KD tissues, 2) screen for additional antigenic targets recognized by the immune response to KD, and 3) evaluate the antibody responses of KD and control sera to KD antigens. Our findings will inform pathogenesis of KD, with the long-term goals of improving diagnosis and treatment of KD, enabling prevention, and reducing healthcare costs from the long-term consequences of coronary artery aneurysms arising in young childhood.

项目摘要/摘要川崎病（KD）是发达国家儿童获得心脏病的主要原因。 KD 可能导致冠状动脉瘤，可能导致终生心脏病，心肌梗死和死亡。临床和流行病学特征支持遗传易感儿童的感染性病因，但是原因已经超过50年了。延迟和错过的诊断增加了冠状动脉动脉瘤。急需诊断测试和改进疗法的发展是取决于识别病因。在初步研究中，我们分析了外周血血浆 KD发作后1-3周的反应使用单细胞RT-PCR，并产生60个单克隆抗体（mab）来自这些浆膜。我们使用这些mAB来确定其靶抗原。我们发现32/60 MAB，源自9/11 kD患者，在纤毛中鉴定出胞质内病毒样体（ICI） KD儿童的支气管上皮，但没有婴儿对照。使用肽阵列识别结合表位，氨基酸取代阵列和ELISA，我们发现32个（16％）mAb中的5个来自3 kD患者使用冠状动脉动脉瘤，识别KD肽表位1。KD肽1完全阻止结合的结合这些对KD ICI的mAb，表明ICI中存在具有结合表位序列的蛋白质。此外，发烧发烧后的5/8 kD患者的治疗前血清>第8天，但婴儿控制血清具有肽表位的IgG抗体。我们最近使用这种方法确定了两个其他表位需要进一步研究。这些发现支持我们的假设，即至少一部分KD病例是由目前未识别的无处不在的病毒病毒剂感染，含有KD肽表位1 顺序。在此提案中，我们假设质子靶向的抗原鉴定了浆膜反应的抗原到KD可以导致病因（IES）的鉴定和KD血清学检查的发展。测试这些假设，我们将：1）确定由我们的KD mAb识别的特定蛋白质与KD结合组织，2）筛选对KD免疫反应识别的其他抗原靶标的，3）评估 KD和控制血清对KD抗原的抗体反应。我们的发现将为KD的发病机理提供信息具有改善KD诊断和治疗的长期目标，使预防和减少医疗保健费用来自童年时期冠状动脉动脉瘤的长期后果。