Assessment Of Patients With Borrelia Infection

莱姆病是一种由伯氏疏螺旋体感染引起的多系统疾病，是美国主要的媒介传播疾病。我们目前的工作涉及莱姆病的以下领域：开发新的感染检测和生物标志物、研究人类伯氏疏螺旋体感染的持续性、寻找南方蜱相关皮疹病 (STARI) 的病因，以及研究免疫反应在莱姆病和 PLDS 中的作用。 莱姆病诊断的主要问题之一是缺乏针对伯氏疏螺旋体的高度特异性和敏感性的检测方法，并且缺乏可用于评估治疗反应的测试。此类检测将极大地促进莱姆病的准确诊断和个体患者对治疗反应的评估。目前还没有这样的测试。我们开发了一种使用荧光素酶免疫沉淀系统 (LIPS) 的新测试，用于分析对一组伯氏疏螺旋体蛋白的抗体反应，以诊断莱姆病。由重复的抗原肽序列组成的合成蛋白（称为 VOVO）具有最佳的诊断性能，类似于 C6 测试（使用我们帮助开发的肽 ELISA 进行的诊断测试，具有高度的敏感性和特异性）。 VOVO LIPS 测试显示出抗体检测的宽动态范围，跨越 10,000 倍以上，无需血清稀释；并为评估莱姆病患者的抗体反应提供了一种有效的定量方法。 最近的研究表明，抗生素治疗后，伯氏疏螺旋体可能会在动物体内持续存在，并且可以通过使用天然蜱载体（肩突硬蜱）通过喂养获得该生物体来检测。这种情况是否发生在人类身上尚不清楚。我们实施了一项新的临床方案来研究该方法在识别治疗的人类莱姆病中伯氏疏螺旋体的持久性方面的效用。 STARI 是一种类似于莱姆病皮疹的皮疹，这种皮疹发生在东南部和中南部各州的居民身上，与孤星蜱 (Amblyomma americanum) 的叮咬有关。皮疹的原因尚不清楚，因为这是疾病的自然病程。我们有一个临床方案来调查 STARI 的病因，并且我们正在应用新的基因组工具，根据 16S rRNA 核糖体基因中的物种特异性序列对 STARI 患者的皮肤活检进行识别细菌。 炎症先天免疫反应对于控制早期播散性感染至关重要，而适应性免疫反应，尤其是体液免疫反应，对于控制组织中的螺旋体水平和动物模型中莱姆关节炎的缓解至关重要。我们正在检查 PLDS 患者和对照者对伯氏疏螺旋体免疫原性显性抗原的抗体反应。进一步研究抗疏螺旋体免疫反应可能有助于阐明 PLDS 的致病机制，并为未来的诊断和治疗方法提供重要信息。 我们有一个临床方案，其中我们使用 DNA 微阵列来表征 EM 个体皮肤活检中的基因表达模式，目的是捕获人类宿主对病原体暴露的反应。我们还在研究以淋巴细胞为主的脑膜炎和以中性粒细胞为主的脑膜炎之间免疫反应的差异。这些研究的结果将成为了解感染的基础生物学的窗口。

Lyme disease is a multisystem illness caused by infection with the spirochete Borrelia burgdorferi and it is the leading vector-borne disease in the United States. Our current work addresses the following areas in Lyme disease: development of new tests and biomarkers for infection, investigation of persistence of infection with B. burgdorferi in humans, search for the cause of Southern Tick-associated Rash Illness (STARI), and investigation of the role of immune response in Lyme disease and PLDS. One of the main problems in Lyme diagnosis has been the lack of highly specific and sensitive assays for B. burgdorferi and the lack of a test that could be used to assess response to therapy. Such assays should greatly facilitate the accurate diagnosis of Lyme disease and assessment of response to therapy in individual patients. Currently, no such test is available. We have developed a new test using the luciferase immunoprecipitation systems (LIPSs) for profiling of the antibody responses to a panel of B. burgdorferi proteins for the diagnosis of Lyme disease. A synthetic protein consisting of a repeated antigenic peptide sequence, named VOVO, had the best diagnostic performance, similar to the C6 test (a diagnostic test using a peptide ELISA that we have helped develop and is highly sensitive and specific). The VOVO LIPS test displays a wide dynamic range of antibody detection spanning over 10,000-fold without the need for serum dilution; and offers an efficient quantitative approach for evaluation of the antibody responses in patients with Lyme disease. Recent studies have shown that B. burgdorferi may persist in animals after antibiotic therapy and can be detected by using the natural tick vector (Ixodes scapularis) to acquire the organism through feeding. Whether this occurs in humans is unknown. We have implemented a new clinical protocol to investigate the utility of this approach for identifying persistence of B. burgdorferi in treated human Lyme disease. STARI is a rash similar to the rash of Lyme disease that occurs in persons residing in southeastern and south-central states and is associated with the bite of the lone star tick, Amblyomma americanum. The cause of the rash is unknown, as it is the natural course of the disease. We have a clinical protocol to investigate the cause of STARI, and we are applying new genomic tools that identify bacteria based on species-specific sequences in the 16S rRNA ribosomal genes to the skin biopsies from patients with STARI. Inflammatory innate immune responses are critical in the control of early disseminated infection, while adaptive immune responses are vitally important, particularly the humoral immune response, in controlling spirochete levels in tissues and resolution of Lyme arthritis in animal models. We are examining the antibody response to immunogenically dominant antigens of B. burgdorferi in PLDS patients and controls. Further investigation of the anti-borrelia immune response may help in elucidating the pathogenic mechanism of PLDS and yield important information for future approaches to diagnosis and treatment. We have a clinical protocol in which we use DNA microarrays to characterize gene expression patterns in skin biopsies from individuals with EM, with the aim of capturing the human host response to pathogen exposure. We are also investigating the differences in immunological response between predominantly lymphocytic meningitis and predominantly neutrophilic meningitis. Results from these studies will serve as a window into the fundamental biology of the infection.