Identifying Kawasaki Disease-Specific Antibodies and Antigens

识别川崎病特异性抗体和抗原

基本信息

批准号：
9932769
负责人：
ANNE H ROWLEY
金额：
$ 23.1万
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：
2018
资助国家：
美国
起止时间：
2018-05-04 至 2020-09-22
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9932769
关键词：
5 year old Acute Adult Affinity Age-Months Aneurysm Antibodies Antibody Affinity Antigen Targeting Antigens Antiviral Agents Antiviral Response Binding Biological Assay Biometry Blood Blood Vessels CD8-Positive T-Lymphocytes Cells Chicago Child Childhood Clinical Cloning Communicable Diseases Complementary DNA Coronary Arteriosclerosis Coronary artery Data Detection Developed Countries Developing Countries Diagnosis Enzyme-Linked Immunosorbent Assay Epidemic Epidemiology Etiology Expression Library Fever Genes Genetic Predisposition to Disease Geography Goals Grant Health Care Costs Heart Diseases Immune Immune response Immunoglobulin A Immunoglobulins Immunohistochemistry Immunology Immunoprecipitation Incidence Inclusion Bodies Individual Infant Infection Infectious Agent Interferon Type I Laboratories Light Lung Lymphoid Mediating Methods Monoclonal Antibodies Morbidity - disease rate Mucocutaneous Lymph Node Syndrome Oligoclonal Bands Patients Pediatric Hospitals Pediatrics Peptide Sequence Determination Pharyngeal structure Plasmablast Preparation Prevention Production Proteins Reporting Reverse Transcriptase Polymerase Chain Reaction Sampling Somatic Mutation Spleen Testing Tissues Transfection Urine Virus Vitelliform macular dystrophy Western Blotting antigen binding base bronchial epithelium cDNA Expression coronary arteritis cytotoxic disorder control expression vector immunoreactivity improved mortality multidisciplinary peripheral blood response synthetic antibodies tool transcriptome virology

项目摘要

Kawasaki Disease (KD) is the leading cause of acquired heart disease in children in developed nations. At Ann & Robert H. Lurie Children's Hospital of Chicago, we diagnose 60-70 new cases/year, and most US children's hospitals diagnose scores of KD patients annually. KD is an acute febrile illness with clinical features commonly observed in infectious diseases, can result in coronary artery aneurysms that can be particularly severe in infants <6 months of age, has highest incidence in young children <5 years of age, is extremely rare in adults, occurs in epidemics with geographic wavelike spread of illness, and only rarely recurs. These specific epidemiologic features of KD are best explained by infection with a ubiquitous infectious agent that is usually asymptomatic, but can result in KD in genetically susceptible children. No known infectious agent is associated with KD. My laboratory reported an antigen-driven arterial IgA immune response in acute KD. In prior studies, we made synthetic antibodies using oligoclonal immunoglobulin α heavy chains and random light chains from KD arterial tissue cDNA. Cognate light and heavy chain pairs could not be determined from the tissue cDNA available to us. Our original KD synthetic antibodies detected antigen in intracytoplasmic inclusion bodies in acute KD but not infant control bronchial epithelium by immunohistochemistry, but were not of sufficient affinity to bind antigen in immunoprecipitation assays. In other studies, we found that type I interferon-induced genes were upregulated in lungs and coronary arteries of KD patients, consistent with an antiviral immune response. Based on these data, we hypothesize that KD is an immune-mediated illness occurring in genetically predisposed children infected with a presently unidentified virus. In this proposal we will use new methods to rapidly clone and produce antigen-specific synthetic antibodies from single cell sorted peripheral blood plasmablasts, allowing identification of cognate light and heavy chains in individual plasmablasts and production of high-affinity antibodies. We plan to prepare a panel of KD-specific monoclonal antibodies and use them to identify KD-specific antigens. In preliminary studies, we identified an oligoclonal, antigen-driven plasmablast response in peripheral blood of an acute KD patient who developed a coronary artery aneurysm. We prepared synthetic antibody using cognate light and heavy chains from the most prevalent oligoclonal plasmablasts from this patient. This antibody binds to antigen in tissues from other KD patients. This proposal includes a multidisciplinary team of experts in KD, virology, immunology, and biostatistics to perform the following specific aims: 1) Identify and clone the peripheral blood plasmablast response in KD, and 2) Identify antigenic targets of oligoclonal KD peripheral blood plasmablasts. Identification of KD-specific antibodies and antigens is an important step toward identifying KD etiology, improving diagnosis and treatment, and enabling prevention, with the goals of reducing pediatric health care costs, morbidity, and mortality from the long-term consequences of coronary artery aneurysms that are acquired during young childhood.

川崎病（KD）是发达国家儿童获得性心脏病的主要原因。在芝加哥 Ann & Robert H. Lurie 儿童医院，我们每年诊断 60-70 个新病例，并且大多数美国患者儿童医院每年诊断出数十名川崎病患者，川崎病是一种具有临床特征的急性发热性疾病。常见于传染病，可导致冠状动脉瘤，特别是 6 个月以下的婴儿病情严重，5 岁以下的幼儿发病率最高，极为罕见在成人中，发生在疾病呈波状传播的流行病中，并且很少复发。川崎病的流行病学特征最好用普遍存在的传染原感染来解释，这种传染原通常是无症状，但可能导致遗传易感儿童患川崎病，目前尚无已知的传染源与之相关。我的实验室报告了急性 KD 中抗原驱动的动脉 IgA 免疫反应。我们使用寡克隆免疫球蛋白 α 重链和随机轻链制备了合成抗体 KD 动脉组织 cDNA 无法从组织 cDNA 中确定同源轻链和重链对。我们原创的 KD 合成抗体可检测细胞质内包涵体中的抗原。免疫组织化学显示急性川崎病，但不是婴儿对照支气管上皮，但没有足够的亲和力在其他研究中，我们发现 I 型干扰素测定诱导基因。 KD 患者的肺部和冠状动脉中表达上调，与抗病毒免疫反应一致。根据这些数据，我们认为川崎病是一种免疫介导的疾病，发生在遗传上。在这项提案中，我们将使用新的方法来预防感染目前未知病毒的儿童。从单细胞分选的外周血中快速克隆并产生抗原特异性合成抗体浆母细胞，允许识别单个浆母细胞中的同源轻链和重链，我们计划制备一组 KD 特异性单克隆抗体。使用它们来识别 KD 特异性抗原在初步研究中，我们鉴定了一种寡克隆、抗原驱动的抗原。患有冠状动脉瘤的急性川崎病患者外周血中浆母细胞反应。我们使用最流行的寡克隆抗体的同源轻链和重链制备了合成抗体该患者的浆母细胞与其他川崎病患者组织中的抗原结合。包括由 KD、病毒学、免疫学和生物统计学领域的多学科专家组成的团队来执行以下具体目标：1) 鉴定并克隆 KD 中的外周血浆母细胞反应，以及 2) 鉴定寡克隆 KD 外周血浆母细胞的抗原靶标的鉴定和 KD 特异性抗体。抗原是识别 KD 病因、改善诊断和治疗以及使预防，目标是从长远来看降低儿科医疗保健费用、发病率和死亡率儿童时期获得的冠状动脉瘤的后果。