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）：莱姆病是由蜱传播的螺旋体伯氏疏螺旋体感染引起的，是美国最常见的媒介传播疾病，每年报告的病例超过 28,000 例。感染可能局限于皮肤或扩散，导致皮肤、心脏、关节和神经系统疾病。螺旋体的 ospC 基因型可能决定其侵袭性和传播倾向。尽管在感染早期阶段使用抗生素可以实现临床治愈，但播散性感染或诊断延迟可能导致大量的发病率和医疗保健支出。及时准确地诊断莱姆病对于优化治疗和预防该病的长期后遗症至关重要。由于受感染组织中发现的螺旋体很少，因此宿主免疫反应为大多数支持莱姆病诊断的商业实验室测试提供了基础。检测伯氏疏螺旋体反应性抗体的血清学测试（ELISA 和免疫印迹）是最广泛使用的测试，但在早期感染中灵敏度和特异性较低，并且在后期可能不确定，特别是在使用抗生素的情况下。此外，目前的莱姆病血清学测试无法区分先前接触伯氏疏螺旋体和活动性感染，并且迄今为止还没有血清学测试可用于评估对治疗的反应。该第一阶段提案旨在通过评估使用全血诊断播散性伯氏疏螺旋体感染和监测治疗反应的新型 T 细胞细胞因子测定的可行性，改进目前可用的莱姆病诊断测试。我们方法的关键是使用两种独特的伯氏疏螺旋体蛋白来建立感染（OspC）和持久性（VlsE）作为细胞因子诱导的抗原刺激。小鼠蜱传感染后一段时间（包括局部感染、传播和持续）将产生 OspC 和 VlsE 的重组形式，并绘制 T 细胞细胞因子谱。将优化抗原，用于检测由代表与传播相关的主要 ospC 基因型（基因型 A、B、I、K 和 N）的 5 种伯氏疏螺旋体感染引起的标志性细胞因子反应。优化抗原的检测将用于评估特征细胞因子的下降速度是否可以预测抗生素消除小鼠感染的情况。最后，将使用患有和不患有莱姆病的受试者的血液样本进行试点研究，以评估反应的特异性。这项一期研究的结果将为开发针对莱姆病的新型快速和特异性细胞免疫测定奠定基础，以补充当前和未来的血清学测试，以增强早期诊断并监测对治疗的反应。 公共卫生相关性：莱姆病是由蜱传螺旋体伯氏疏螺旋体感染引起的，是美国最常见的媒介传播疾病。及时诊断感染对于获得最佳治疗反应和预防长期后遗症非常重要。该项目将评估 T 细胞细胞因子测定在检测播散性伯氏疏螺旋体感染和监测治疗效果方面的效用。