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（由适用提供）：囊性纤维化是美国最常见的致命遗传疾病，其特征是气道慢性感染。两种基于培养的方法以及非文化的方法（16S测序）鉴定出的最常见的生物是铜绿假单胞菌和金黄色葡萄球菌。除了这些细菌外，烟曲霉的真菌感染也很常见。尽管定殖是常见的 - 细菌或真菌的侵入性感染很少见。这与非CF种群中铜绿假单胞菌和金黄色葡萄球菌的急性感染相反，无论它们是社区获得的还是医院感染。长期以来，人们已经认识到，具有多克隆IgG反应的CF患者具有强大的适应性免疫反应以及对铜绿假单胞菌的T细胞增殖剂反应。我们已经在CF患者的肺和支气管刷中确定了强烈的B细胞反应。我们将检验以下假设：这些粘膜B细胞用以下目的编码病原体特异性免疫反应：特定目标1。创建CF肺的人类单克隆抗体小组。 B细胞将从临床支气管镜或接受移植的CF患者中获得。 B细胞将用爱泼斯坦Barr病毒（EBV）转化，然后融合到骨髓瘤融合伴侣。然后，如前所述，我们将将Ig同型表征为体细胞超女的程度。特定目的2。测试人单克隆抗体的抗原特异性。我们首先测试了从患者配对的BAL样品中对临床微生物学实验室中获得的病原体的抗体，并测试针对铜绿假单胞菌，嗜性嗜性嗜性菌群，金黄色葡萄球菌（MSSA和MRSA）（MSSA和MRSA）和A. fumigatus的一般反应性。交叉反应性将通过Elsia和Western印迹分析进行评估。反应与铜绿假单胞菌反应的克隆将使用中性小鼠模型在肺部感染的体内动物中进行测试。