T cell cytokine assay for the diagnosis of disseminated Lyme borreliosis

T 细胞细胞因子测定用于诊断播散性莱姆疏螺旋体病

• 批准号: 8058201
• 负责人: Linda K. Bockenstedt
• 金额: $ 28.1万
• 依托单位: L2 DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-01 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8058201
• 关键词: Acute; Affect; Antibiotics; Antibodies; Antibody Formation; Antigens; Asia; Bacterial Infections; Biological Assay; Bite; Blood specimen; Borrelia burgdorferi; Case Study; Centers for Disease Control and Prevention (U.S.); Clinical; Complement; DNA; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Disease; Doxycycline; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Europe; Exanthema; Exposure to; Focal Infection; Future; Genotype; Geographic Distribution; Health Expenditures; Heart; Human; Immune; Immune response; Immunoblotting; Incidence; Infection; Infectious Skin Diseases; Inflammatory Response; Ixodes; Joints; Kinetics; Laboratories; Lead; Lyme Disease; Maps; Measures; Medical; Methods; Modeling; Monitor; Morbidity - disease rate; Mus; Nervous system structure; North America; Order Spirochaetales; OspC protein; Patients; Peptides; Phase; Pilot Projects; Prevention strategy; Production; Proteins; Public Health; Recombinant Proteins; Recombinants; Recording of previous events; Regimen; Rheumatoid Arthritis; Sensitivity and Specificity; Serologic tests; Serological; Skin; Specificity; Staging; Stimulus; Systemic infection; T cell response; T-Lymphocyte; Technology; Testing; Tick Infestations; Tick-Borne Infections; Ticks; Tissues; Treatment Efficacy; United States; Vector-transmitted infectious disease; Whole Blood; base; commercialization; cytokine; experience; human subject; improved; microorganism culture; novel; phase 1 study; phase 2 study; prevent; prototype; response; tick borne spirochete

DESCRIPTION (provided by applicant): Lyme disease, due to infection with the Ixodes tick-transmitted spirochete Borrelia burgdorferi, is the most common vector-borne disease in the United States, with more than 28,000 cases reported annually. The infection can remain localized to the skin or disseminate to cause disease in the skin, heart, joints and nervous system. The ospC genotype of the spirochete may determine its invasiveness and propensity to disseminate. Although antibiotics achieve clinical cure when administered in early stages of infection, disseminated infection or delay in diagnosis can lead to substantial morbidity and health care expenditures. Timely and accurate diagnosis of Lyme disease is essential for optimizing treatment and for preventing long-term sequelae of the disease. Because few spirochetes are found in infected tissues, the host immune response provides the basis for most commercial laboratory tests that support a diagnosis of Lyme disease. Serologic tests (ELISA and immunoblot) that detect B. burgdorferi-reactive antibodies are the most widely used tests, but have lower sensitivity and specificity in early infection and can be indeterminate in later stages, particularly if antibiotics have been administered. In addition, current Lyme serologic tests do not distinguish previous exposure to B. burgdorferi from active infection, and no serologic test to date can be used to assess response to therapy. This Phase 1 proposal seeks to improve upon the currently available Lyme diagnostic tests by evaluating the feasibility of a novel T cell cytokine assay using whole blood for the diagnosis of disseminated B. burgdorferi infection and for monitoring response to therapy. The key to our approach is the use of two unique B. burgdorferi proteins required for establishment of infection (OspC) and persistence (VlsE) as antigenic stimuli for cytokine induction. Recombinant forms of OspC and VlsE will be produced and T cell cytokine profiles mapped after tick-borne infection in mice over a period encompassing localized infection, dissemination and persistence. Antigens will be optimized for detection of signature cytokine responses elicited by B. burgdorferi infection with 5 strains representing the main ospC genotypes associated with dissemination (genotypes A, B, I, K, and N). Assays with optimized antigens will be used to assess whether the rate of decline of signature cytokines predicts antibiotic elimination of infection in mice. Finally, pilot studies will be performed to assess the specificity of the response using blood samples from subjects with and without Lyme disease. The results of this Phase 1 study will set the stage for the development of new rapid and specific cellular immune assays for Lyme disease that complement current and future serologic tests to enhance early diagnosis and to monitor response to therapy. PUBLIC HEALTH RELEVANCE: Lyme disease, due to infection with the tick-borne spirochete Borrelia burgdorferi, is the most common vector-borne disease in the United States. Timely diagnosis of infection is important for optimal response to therapy and to prevent long-term sequelae. This project will assess the utility of a T cell cytokine assay for the detection of disseminated B. burgdorferi infection and for monitoring efficacy of treatment.

描述（由申请人提供）：莱姆病由于感染了ixodes tick tick tick的螺旋体Borrelia burgdorferi，是美国最常见的媒介传播疾病，每年报告超过28,000例。感染可以保持位于皮肤上或传播以引起皮肤，心脏，关节和神经系统的疾病。螺旋体的OSPC基因型可能决定其侵入性和传播倾向。尽管在感染的早期阶段进行抗生素时，抗生素可实现临床治疗，但散布感染或诊断延迟会导致大量的发病率和医疗保健支出。及时，准确的莱姆病诊断对于优化治疗和预防疾病的长期后遗症至关重要。由于在感染组织中发现螺旋体很少，因此宿主免疫反应为大多数支持诊断莱姆病的商业实验室测试提供了基础。检测B. burgdorferi反应性抗体的血清学测试（ELISA和免疫印迹）是使用最广泛的测试，但在早期感染中具有较低的敏感性和特异性，并且在较晚的阶段可能不确定，尤其是在抗生素中施用。此外，当前的莱姆血清学检查并不能区分先前接触B. burgdorferi与主动感染，并且迄今为止尚无血清学检查可用于评估对治疗的反应。该第1阶段的建议旨在通过评估新型T细胞细胞因子测定的可行性，利用全血为诊断散布的B. burgdorferi感染并监测治疗反应，以改善当前可用的莱姆诊断测试。我们方法的关键是使用两种独特的B. burgdorferi蛋白来建立感染（OSPC）和持久性（VLSE）作为细胞因子诱导的抗原刺激。将产生OSPC和VLSE的重组形式，并在小鼠感染后在小鼠感染后映射的T细胞细胞因子特征，其中包括局部感染，传播和持久性。抗原将被优化，以检测B. burgdorferi感染引起的签名细胞因子反应，并具有5​​种代表与传播相关的主要OSPC基因型（基因型A，B，I，K和N）的菌株。具有优化抗原的测定将用于评估签名细胞因子的下降速率是否预测了小鼠感染的抗生素消除。最后，将使用来自有或没有莱姆病的受试者的血液样本来评估反应的特异性。这一第一阶段研究的结果将为莱姆病的新快速和特定细胞免疫测定奠定基础，这些莱姆病疾病补充了当前和未来的血清学检查，以增强早期诊断并监测对治疗的反应。 公共卫生相关性：莱姆病由于与tick传播的螺旋体玻璃体伯氏菌（Borrelia Burgdorferi）感染，是美国最常见的媒介传播疾病。及时诊断感染对于对治疗的最佳反应和预防长期后遗症很重要。该项目将评估T细胞细胞因子测定的效用，以检测散布的B. burgdorferi感染和监测治疗功效。