Assessment Of Patients With Lyme Infection

莱姆病是由伯氏疏螺旋体感染引起的一种多系统疾病，是美国主要的媒介传播疾病。莱姆病在欧洲(主要是中欧和斯堪的纳维亚半岛)也很常见，在俄罗斯、中国和日本也有发生。在人类中，伯氏杆菌感染主要导致皮肤、神经系统、心脏和关节的感染。莱姆病通常可以用抗生素治疗成功，早期疾病的患者效果最好。不幸的是，一些患者可能对治疗没有完全的反应。我们目前有两个研究莱姆病患者的临床方案。这两项协议都是自然历史研究，并作为多条调查路线的基础。一种方案针对治疗后莱姆病综合征患者。另一种方案允许对典型莱姆病患者进行研究。我们的工作涉及莱姆病的三个方面：实验室诊断、临床表现和对伯氏杆菌的免疫反应。关于实验室诊断学，我们一直专注于开发更好的诊断和感染持久性测试。我们与杜兰大学医学中心的Mario Philipp博士和他的团队合作开发了C6多肽酶联免疫吸附试验。这项检测操作简单，具有高度的敏感性和特异性。这种检测的一个重要优点是，它可以用来诊断接种了莱姆病疫苗的患者的莱姆病，而且可以在欧洲使用，在欧洲，莱姆病可能是由加里尼氏杆菌和阿夫泽利杆菌引起的。我们还与疾病预防控制中心合作，评估了C6抗体反应，以及对其他重组抗原和合成肽的抗体反应，并与两级检测进行了比较。当单独使用时，C6酶联免疫吸附试验对双抗体夹心试验具有相似的敏感性。此外，当与大多数OSPC蛋白(PepC10)C末端发现的10个氨基酸多肽的IgM抗体检测结合使用时，该组合对诊断急性莱姆病患者暴露于伯氏杆菌的敏感性明显更高。除了使用C6抗体反应诊断莱姆病外，我们目前正在评估这种反应作为感染清除的可能标记物。在免疫应答方面，我们与NINDS神经免疫学分会的Roland Martin博士合作，使用一种新的方法来解密T细胞克隆的抗原特异性，研究了伯氏疏螺旋体特异性T细胞克隆的特异性和功能。在这项研究的后续工作中，我们开发了一种高度特异和敏感的技术，通过实时RT-PCR检测和定量T细胞受体(TCR)α或β链互补决定区3转录产物来跟踪单个T细胞克隆。我们检测了神经系统疾病病程中外周血和脑脊液中候选致病T细胞克隆的频率。我们对研究伯氏杆菌感染的先天免疫反应也非常感兴趣。我们使用寡核苷酸芯片检测了人外周血单个核细胞对伯氏杆菌裂解物的转录图谱，作为建立关于莱姆病分子发病机制的新假说的基础。关于临床表现，我们对患者队列中的听力学系统进行了评估。这些评估包括对PureTone敏感性测量、语音接收阈值、语音识别、语音耐受性的研究，以及对高强度呈现水平下语音功能的其他研究。它还包括对中耳完整性和耳声发射的生物力学研究，以识别(或排除)耳蜗损伤，以及评估听觉脑干反应，以识别或排除对上行听觉脑干通路的损伤。

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. Lyme disease is also common in Europe (mainly middle Europe and Scandinavia) and also occurs in Russia, China and Japan. In humans, B. burgdorferi infection causes infection primarily in the skin, nervous system, heart and joints. Lyme disease can usually be treated successfully with antibiotic therapy, with the best results seen in patients with early disease. Unfortunately, some patients may not have a complete response to therapy. We currently have two clinical protocols studying patients with Lyme disease. Both protocols are natural history studies and serve as the basis for multiple lines of investigation. One protocols addresses patients with post treatment Lyme disease syndrome. The other protocol allow for the study of patients with classical Lyme disease. Our work has addressed 3 areas in Lyme disease: laboratory diagnosis, clinical manifestations and immunological responses to B. burgdorferi. Regarding laboratory diagnostics, we have focused in developing better tests for both diagnosis and for persistence of infection. We collaborate with Dr. Mario Philipp and his group at Tulane University Medical Center, in the development of the C6 peptide ELISA. This test is simple to perform and is highly sensitive and specific. An important advantage of this test is that it can be used to diagnose Lyme disease in patients who have received the Lyme disease vaccine, and it can be used in Europe, where Lyme disease may be caused by B. garinii and B. afzelli. We also collaborated with the CDC to evaluate the C6 antibody response, as well as the antibody response to other recombinant antigens and synthetic peptides, in comparison with the 2-tier testing. When used alone, the C6 ELISA had similar sensitivity to 2-tier test. Moreover, when used in combination with the detection of IgM antibodies to a ten-amino acid peptide found at the C-terminus of most OspC proteins (pepC10), the combination had significantly higher sensitivity for diagnosing exposure to B. burgdorferi in patients with acute Lyme disease. Besides the use of the C6 antibody response for diagnosis of Lyme disease, we are currently evaluating this response as a possible marker for clearance of infection. Regarding immunological responses, in collaboration with Dr. Roland Martin (Neuroimmunology Branch, NINDS), we are studying the specificity repertoires and function of Borrelia burgdorferi-specific T cell clones using a novel methodology to decrypt the antigen specificity of T cell clones. In follow up to this study, we developed a highly specific and sensitive technique to track single T cell clones through the detection and quantification of T cell receptor (TCR) alpha or beta chain complementarity-determining region 3 transcripts by real-time RT-PCR. We examined the frequency of the candidate pathogenic T cell clones in the peripheral blood and cerebrospinal fluid (CSF) during the course of neurological disease. We are also very interested in investigating the innate immune response to B. burgdorferi infection. We have examined the transcription profile of human peripheral blood mononuclear cells in response to B.burgdorferi lysate using oligonucleotide microarrays, as a basis for establishing new hypotheses regarding the molecular pathogenesis of Lyme disease. Regarding clinical manifestations, we have evaluated the audiological system in our patient cohort. These evaluations included studies with puretone sensitivity measurements, speech reception threshold, speech recognition, tolerance for speech and other studies of speech functions at high intensity presentation levels. It also included biomechanical studies of middle ear integrity and otoacoustic emissions to identify (or rule out) cochlear damage, and assessment of the auditory brainstem response to identify or rule out damage to the ascending auditory brainstem pathways.