T cell cytokine assay for the diagnosis of disseminated Lyme borreliosis

描述（由申请方提供）：莱姆病是由硬蜱传播的螺旋体伯氏疏螺旋体感染引起的，是美国最常见的病媒传播疾病，每年报告的病例超过28，000例。感染可以局限于皮肤或传播，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。虽然抗生素在感染的早期阶段使用时可以达到临床治愈，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疾病的长期后遗症至关重要。由于在感染的组织中很少发现螺旋体，因此宿主的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B的血清学试验（ELISA和免疫印迹）。伯氏反应性抗体是最广泛使用的测试，但在早期感染中具有较低的灵敏度和特异性，并且在后期阶段可能不确定，特别是如果已经施用抗生素。此外，目前的莱姆病血清学检测不能区分既往接触过B。迄今为止，没有血清学试验可用于评估对治疗的反应。该I期提案旨在通过评估使用全血诊断播散性B的新型T细胞细胞因子测定的可行性来改进目前可用的莱姆病诊断测试。用于监测对治疗的反应。我们的方法的关键是使用两个唯一的B。建立感染（OspC）和持续感染（VlsE）所需的伯氏螺旋体蛋白作为细胞因子诱导的抗原刺激物。将产生OspC和VlsE的重组形式，并在小鼠中蜱传感染后在包括局部感染、传播和持续的一段时间内绘制T细胞细胞因子谱。抗原将被优化用于检测由B引发的标志性细胞因子应答。用代表与播散相关的主要ospC基因型（基因型A、B、I、K和N）的5种菌株检测伯氏螺旋体感染。使用优化抗原的测定将用于评估特征细胞因子的下降速率是否预测小鼠中感染的抗生素消除。最后，将进行初步研究，以使用来自患有和不患有莱姆病的受试者的血液样本评估反应的特异性。这项1期研究的结果将为莱姆病新的快速和特异性细胞免疫检测的开发奠定基础，这些检测将补充当前和未来的血清学检测，以加强早期诊断和监测对治疗的反应。 公共卫生相关性：莱姆病，由于感染蜱传螺旋体伯氏疏螺旋体，是最常见的病媒传播的疾病在美国。及时诊断感染对于治疗的最佳反应和预防长期后遗症非常重要。本项目将评估T细胞细胞因子检测播散性B的实用性。用于监测治疗效果。

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.