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，由于感染蜱传播螺旋体伯氏疏螺旋体，是一种多系统疾病 可表现为局部皮肤感染或累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。体液免疫是一个关键的宿主 对B的防御血液中强烈的早期B细胞反应与 症状消退。B。伯氏菌感染还可引发自身抗体产生和免疫失调 与自身免疫性疾病有关尽管抗体产生对宿主防御的重要性， 已知它们在皮肤中的作用，蜱虫传播的螺旋体首先在皮肤中建立感染。我们应用 先进的转录组技术来研究原发性红斑中发现的稀疏B细胞群 移行性病变和非病变皮肤的受试者谁提出了局限性或传播性莱姆病。 使用10 X单细胞免疫库和基因表达谱，我们鉴定了来自 B细胞具有对免疫攻击（例如感染）的应答特征（即克隆性 扩增、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，在同一个病人的血液中也发现了。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）迄今已发现 2株单抗可与15- 20 kD Bb抗原结合。在本建议中，我们将扩大 重组mAb的产生并确定mAb是否与B反应。burgdorferi或自身抗原。我们将使用 新开发的创新型快速外膜蛋白质组抗体分析（REAP）发现工具， 重组mAb和免疫血清针对外蛋白质组中蛋白质的反应性的通量筛选。 在诊断时和恢复期的几个时间点从莱姆受试者中前瞻性采集的存档血清 随着时间的推移，将筛选针对鉴定的Bb抗原以及自身抗原的反应性。的结果 这些研究将是对早期莱姆病中B细胞抗原特异性的首次组织特异性分析， 并提供对免疫反应的持久性和抗体影响感染的能力的深入了解 结果。它还可以提供新的靶抗原的身份，用于早期莱姆病的诊断测试 或将新感染与先前治疗的感染区分开。此外，这些研究可能会提供洞察力， 当自反应性在B之后首次出现时。伯氏螺旋体感染和对靶向 抗原这些研究的成功完成将为研究皮肤B细胞反应建立模型 其他具有公共卫生意义的病媒传播病原体。