Immunophenotypic analysis of the cutaneous humoral response in early Lyme disease

早期莱姆病皮肤体液反应的免疫表型分析

基本信息

批准号：
10561695
负责人：
Linda K. Bockenstedt
金额：
$ 20.94万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-03 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10561695
关键词：
Adverse event Aftercare Antibiotics Antibodies Antibody Formation Antibody Response Antigen Targeting Antigens Archives Autoantibodies Autoantigens Autoimmune Autoimmune Diseases Autoimmunity Autologous B-Cell Antigen Receptor B-Lymphocytes B-cell receptor repertoire sequencing Blood Blood specimen Borrelia burgdorferi COVID-19 Cell Separation Cells Characteristics Clinical Clonal Expansion Cognitive Communicable Diseases Complex Convalescence Custom Cutaneous Data Detection Diagnosis Diagnostic tests Disease Disease Outcome Engineering Evolution Fatigue Future Gene Expression Gene Expression Profiling Heart Host Defense Human Humoral Immunities Immune Immune Sera Immune response Immunity Immunoglobulin A Immunoglobulin Class Switching Immunoglobulin G Immunoglobulin M Immunoglobulin Somatic Hypermutation Immunophenotyping In Vitro Infection Infectious Skin Diseases Investigation Joints Knowledge Lesion Localized Skin Lesion Lyme Arthritis Lyme Disease Mediating Memory Modality Modeling Monoclonal Antibodies Musculoskeletal Pain Mutate Nervous System Order Spirochaetales Outcome Pathogenesis Pathogenicity Pathology Patients Phenotype Plasma Cells Play Population Post Treatment Lyme Disease Syndrome Production Proteins Public Health Reaction Recombinants Reporting Research Resolution Role Sampling Serology Site Skin Specificity Strategic Planning Surface Symptoms System Systems Biology Technology Testing Tick-Borne Diseases Tick-Borne Infections Time Tissue Sample Tissues United States United States National Institutes of Health Vector-transmitted infectious disease adverse outcome autoimmune arthritis autoreactivity body system design erythema migrans experience experimental study extracellular high throughput screening human monoclonal antibodies improved innate immune mechanisms innovation insight joint inflammation monoclonal antibody production new technology novel pathogen prospective response single-cell RNA sequencing skin lesion tick bite tick transmission tool transcriptome vector-borne pathogen

项目摘要

PROJECT SUMMARY Lyme disease, due to infection with the tick-transmitted spirochete Borrelia burgdorferi, is a multisystem disorder that can present with either localized skin infection or with disseminated infection involving multiple skin sites and other organs systems, including the heart, nervous system and joints. Humoral immunity is a critical host defense against B. burgdorferi infection, and a strong early B cell response in the blood is associated with symptom resolution. B. burgdorferi infection can also trigger autoantibody production and immune dysregulation associated with autoimmune pathologies. Despite the importance of antibody production to host defense, little is known about their role in the skin where tick-transmitted spirochetes first establish infection. We have applied advanced transcriptome technologies to study the sparse B cell populations found in the primary erythema migrans lesion and nonlesional skin of subjects who presented with localized or disseminated Lyme disease. Using 10X single cell immune repertoire and gene expression profiling, we identified antibody sequences from B cells that have the characteristics of a response to an immune challenge such as infection (i.e. clonal expansion, somatic hypermutation and class switching). These sequences were not present in nonlesional skin, but in some cases were also found in the blood of the same patient. In preliminary data, we have produced 11 recombinant human monoclonal antibodies (mAb) based on these antibody sequences and to date, have found that 2 mAb react with a surface exposed Bb antigen, 15-20kD in size. In this proposal, we will expand the recombinant mAb production and determine if the mAb react with B. burgdorferi or self-antigens. We will use the newly developed and innovative Rapid Exoproteome Antibody Profiling (REAP) discovery tool for high throughput screening of recombinant mAb and immune sera for reactivity against proteins in the exoproteome. Archived sera collected prospectively from Lyme subjects at diagnosis and several time points in convalescence will be screened for reactivity against identified Bb antigens as well as self antigens over time. The results of these studies will be the first tissue-specific analysis of the antigen specificity of B cells in early Lyme disease, and provide insight into the durability of immune responses and capacity of the antibodies to influence infection outcomes. It may also provide identities of new target antigens for use in diagnostic tests for early Lyme disease or distinguish new infection from previously treated infections. In addition, these studies may provide insight into when self-reactivity first arises after B. burgdorferi infection and the evolution of the response to targeted antigens. Successful completion of these studies will create a model for investigation of the skin B cell response to other vector-borne pathogens of public health significance.

项目概要莱姆病是由蜱传播的螺旋体伯氏疏螺旋体感染引起的，是一种多系统疾病可能表现为局部皮肤感染或涉及多个皮肤部位的播散性感染以及其他器官系统，包括心脏、神经系统和关节。体液免疫是关键宿主防御伯氏疏螺旋体感染，血液中强烈的早期 B 细胞反应与症状解决。伯氏疏螺旋体感染也会引发自身抗体的产生和免疫失调与自身免疫病理有关。尽管抗体的产生对于宿主防御很重要，但人们对此却知之甚少。了解它们在皮肤中的作用，蜱传播的螺旋体首先在皮肤中建立感染。我们已经申请了先进的转录组技术用于研究原发性红斑中发现的稀疏 B 细胞群患有局部或播散性莱姆病的受试者的移行性病变和非病变皮肤。使用 10X 单细胞免疫库和基因表达谱，我们鉴定了来自 B 细胞具有对感染等免疫挑战做出反应的特征（即克隆扩展、体细胞超突变和类别转换）。这些序列不存在于非病变皮肤中，但在某些情况下，也在同一患者的血液中发现了这种物质。在初步数据中，我们已经生产了 11 个基于这些抗体序列的重组人单克隆抗体（mAb）迄今为止已发现 2 mAb 与表面暴露的 Bb 抗原（大小为 15-20kD）发生反应。在本提案中，我们将扩大重组 mAb 的产生并确定 mAb 是否与伯氏疏螺旋体或自身抗原发生反应。我们将使用新开发的创新型快速外蛋白组抗体分析 (REAP) 发现工具，用于高对重组单克隆抗体和免疫血清进行通量筛选，以检测其与外蛋白质组中蛋白质的反应性。前瞻性地从莱姆病受试者诊断时和恢复期的几个时间点采集的存档血清随着时间的推移，将筛选针对已识别的 Bb 抗原以及自身抗原的反应性。结果这些研究将是对早期莱姆病 B 细胞抗原特异性的首次组织特异性分析，并深入了解免疫反应的持久性和抗体影响感染的能力结果。它还可以提供用于早期莱姆病诊断测试的新靶抗原的鉴定或区分新感染和先前治疗过的感染。此外，这些研究可能会提供深入了解当伯氏疏螺旋体感染后首次出现自身反应时，以及对目标反应的演变抗原。这些研究的成功完成将为研究皮肤 B 细胞反应创建一个模型其他具有公共卫生意义的媒介传播病原体。