Generation of Novel Human Monoclonals for Lung Disease

用于肺部疾病的新型人单克隆抗体的产生

基本信息

批准号：
9128312
负责人：
JAY K KOLLS
金额：
$ 19.25万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9128312
关键词：
Acute Allergic Bronchopulmonary Aspergillosis Animals Antibiotics Antibodies Antigens Aspergillus fumigatus Asthma B-Lymphocytes Bacteremia Bacteria Binding Bronchi Bronchoscopy Cells Child Chronic Chronic Obstructive Airway Disease Clinical Clinical Microbiology Communities Cystic Fibrosis Data Disease Enzyme-Linked Immunosorbent Assay Epithelial Cells Epithelium Fungemia Generations Genes Hereditary Disease Human Human Herpesvirus 4 Immune response Immune system Immunoglobulin G Immunoglobulin Genes Immunoglobulin Somatic Hypermutation In Vitro Infection Infection Control Inflammation Interferons Interleukin-13 Interleukin-17 Interleukin-2 Life Lung Lung Transplantation Lung diseases Mediating Methods Mucosal Immune Responses Multiple Myeloma Mutation Mycoses Nosocomial Infections Organism Patients Plasma Cells Population Proteins Pseudomonas aeruginosa Recruitment Activity Refractory Role Sampling Specificity Staging Staphylococcus aureus Stenotrophomonas maltophilia Steroids T-Cell Receptor T-Lymphocyte TNFSF5 gene Testing Th2 Cells Tonsil Transplantation Western Blotting Work adaptive immunity airway inflammation base cohort cross reactivity cystic fibrosis patients cytokine design end stage disease human monoclonal antibodies in vivo innovation interleukin-22 methicillin resistant Staphylococcus aureus mouse model mutant novel novel therapeutics pathogen peripheral blood prevent public health relevance receptor research study response transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): Cystic Fibrosis is the most common lethal genetic disease in the US and is characterized by chronic infection inflammation in the airway. The most common organisms identified by both culture-based methods as well as non-culture based methods (16s sequencing) are Pseudomonas aeruginosa and Staphylococcus aureus. In addition to these bacteria, fungal infection with Aspergillus fumigatus is also common. Although colonization is common - invasive infection with bacteremia or fungemia is rare. This is in contrast to acute infections with P. aeruginosa and S. aureus in non-CF populations whether they are community acquired or nosocomial infections. It has long been recognized that there is strong adaptive immune response in patients with CF characterized by polyclonal IgG responses as well as T-cell proliferative responses to P. aeruginosa. We have identified a strong B-cell response in the lungs and bronchial brushes of CF patients. We will test the hypothesis that these mucosal B-cells encode pathogen specific immune responses with the following Aims: Specific Aim 1. Create a panel of human monoclonal antibodies from the CF lung. B-cells will be obtained from clinical bronchoscopies or from CF patients undergoing transplant. B-cells will be transformed with Epstein Barr Virus (EBV) and then fused to a myeloma fusion partner. We will then characterize Ig isotype as the extent of somatic hyper-mutation as previously described. Specific Aim 2. Test the antigen specificity of the human monoclonal antibodies. We first test the antibodies against the pathogens obtained in the clinical microbiology lab from the paired BAL sample of the patient as well as test for general reactivity against lab strains of P. aeruginosa, Stenotrophomonas maltophilia, S. aureus (both MSSA and MRSA), and A. fumigatus. Cross-reactivity will be assessed by both ELSIA and Western blot analysis. Clones that react against P. aeruginosa will be tested in an in vivo animal of pulmonary infection using the neutropenic mouse model.

描述（由申请人提供）：囊性纤维化是美国最常见的致命遗传性疾病，其特征是气道中的慢性感染炎症，是通过基于培养的方法和基于非培养的方法鉴定的最常见的生物体。 16s 测序）是铜绿假单胞菌和金黄色葡萄球菌除了这些细菌之外，烟曲霉的真菌感染也很常见，尽管定植是常见的 - 侵袭性的。与非 CF 人群中的铜绿假单胞菌和金黄色葡萄球菌急性感染相比，无论是社区获得性感染还是医院感染，菌血症或真菌血症感染都很罕见。人们早已认识到，在非 CF 人群中存在强烈的适应性免疫反应。以多克隆 IgG 反应以及对铜绿假单胞菌的 T 细胞增殖反应为特征的 CF 患者，我们在肺和支气管刷中发现了强烈的 B 细胞反应。我们将测试这些粘膜 B 细胞编码病原体特异性免疫反应的假设，目的如下：具体目标 1. 从临床支气管镜检查或获得的 CF 肺部 B 细胞中创建一组人单克隆抗体。来自接受移植的 CF 患者的 B 细胞将被 Epstein Barr 病毒 (EBV) 转化，然后与骨髓瘤融合伴侣融合。具体目标 2. 测试人单克隆抗体的抗原特异性我们首先测试在临床微生物学实验室中从患者的配对 BAL 样本中获得的针对病原体的抗体。对于铜绿假单胞菌、嗜麦芽寡养单胞菌、金黄色葡萄球菌（MSSA 和 MRSA）和烟曲霉实验室菌株的一般反应性将是。通过 ELSIA 和蛋白质印迹分析进行评估，将使用中性粒细胞减少小鼠模型在肺部感染动物体内测试对铜绿假单胞菌有反应的克隆。