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

• 批准号: 10451111
• 负责人: Linda K. Bockenstedt
• 金额: $ 25.13万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 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/10451111
• 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; 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; In Vitro; Infection; Infectious Skin Diseases; Investigation; Joints; Knowledge; Lead; Lesion; Localized Skin Lesion; Lyme Arthritis; Lyme Disease; Mediating; Memory; Modality; Modeling; Monoclonal Antibodies; Musculoskeletal Pain; Mutate; Nervous system structure; Order Spirochaetales; Outcome; Pathogenesis; Pathogenicity; Pathology; Patients; Phenotype; Plasma Cells; Play; Population; Production; Proteins; Public Health; Recombinants; Reporting; Research; Resolution; Role; Sampling; Serology; Site; Skin; Specificity; Strategic Planning; Surface; Symptoms; Syndrome; 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; base; 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

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.

项目摘要 莱姆病由于感染了tick虫传播的螺旋体玻璃体burgdorferi，是一种多系统疾病 可能出现局部皮肤感染或涉及多个皮肤部位的传播感染 以及其他器官系统，包括心脏，神经系统和关节。体液免疫是关键主持人 防御爆发芽孢杆菌感染以及血液中强烈的早期B细胞反应与 症状解决。 B. Burgdorferi感染还可以触发自身抗体的产生和免疫失调 与自身免疫性病理相关。尽管抗体生产对托管防御的重要性很重要，但几乎没有 知道它们在皮肤中的作用，在皮肤上首先建立感染。我们已经申请了 先进的转录组技术，研究了主要红斑中发现的稀疏B细胞种群 出现局部或传播莱姆病的受试者的迁移病变和非静态皮肤。 使用10倍单细胞免疫库和基因表达分析，我们从 具有对免疫挑战（例如感染）反应特征的B细胞（即克隆 膨胀，体外夸张和班级切换）。这些序列不存在于非平静的皮肤中， 但是在某些情况下也发现了同一患者的血液。在初步数据中，我们生产了11 基于这些抗体序列的重组人单克隆抗体（MAB）已发现 该2个mAb与表面裸露的BB抗原反应，大小为15-20kD。在此提案中，我们将扩大 重组MAB的产生，并确定MAB是否与B. burgdorferi或自我抗原反应。我们将使用 新开发和创新的快速杂蛋白抗体分析（REAP）发现工具 重组MAB和免疫血清的吞吐量筛选，以使蛋白质中的蛋白质反应性。 诊断时从莱姆受试者和康复中几个时间点收集的存档血清 随着时间的流逝，将筛选针对已鉴定的BB抗原以及自我抗原的反应性。结果 这些研究将是对早期莱姆病的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耐用性和抗体影响感染的能力 结果。它还可能提供新的靶抗原的身份，用于用于早期莱姆病的诊断测试 或区分新感染与先前治疗的感染。此外，这些研究可能会洞悉 当自反应性在B. burgdorferi感染后首次产生时，并且对靶向的反应的演变 抗原。这些研究的成功完成将创建一个模型，以研究皮肤B细胞反应 对于其他具有公共卫生意义的媒介传播病原体。