Diagnosis and Pathogenesis of Early Lyme Disease

描述（由申请人提供）：本拨款提案的目标是改进莱姆病的诊断测试，并确定导致更严重疾病的螺旋体和宿主因素。这一目标将通过分析来自大型莱姆病患者的回顾性和前瞻性样本来实现。在本项目中，将前瞻性地评估第二代主要候选血清学检测IgM OspC （C10）肽ELISA和IgG VIsE （IR6）肽ELISA，并将其与早期或晚期莱姆病患者和对照组的全细胞超声ELIoA和Western blot结果进行比较。研究人员将探索对伯氏疏螺旋体新型糖脂抗原的抗体反应，以了解抗生素治疗后这些反应的系列测定是否可以作为螺旋体杀伤的潜在替代标志物。将定量PCR与标准PCR和培养相比较，确定其作为莱姆病患者诊断试验的价值。将描述移徙性红斑（EM）皮肤病变的皮肤活检样本中的螺旋体负荷和这些病变分离的螺旋体毒力因子，并将结果与螺旋体传播和感染严重程度的临床标志物相关联。最后，将确定这些患者EM病变中的细胞因子和趋化因子谱，并将其与螺旋体传播和感染严重程度的临床标志物以及细胞和体液免疫功能的血液检测结果相关联。这些研究可能导致莱姆病血清诊断、直接检测螺旋体和评估螺旋体杀伤的更准确检测。此外，更多地了解与严重疾病相关的螺旋体毒力和宿主因素，可能有助于为患者开发实验室标记物，这些患者将受益于更强化的治疗。

DESCRIPTION (provided by applicant): The goal of this grant proposal is to improve diagnostic testing for Lyme disease and to identify spirochetal and host factors that lead to more severe disease. This goal will be carried out by the analysis of samples from large retrospective and prospective series of well-characterized patients with Lyme disease. For this project, a leading candidate second-generation serologic test, an IgM OspC (C10)-peptide ELISA and an IgG VIsE (IR6)-peptide ELISA, will be assessed prospectively' and compared with the results obtained by whole-cell sonicate ELIoA and Western blot in patients with early or late Lyme disease and in control subjects. Antibody responses to novel glycolipid antigens of B. burgdorferi will be explored to learn whether serial determinations of these responses after antibiotic treatment may serve as a potential surrogate marker for spirochetal killing. The value of quantitative PCR as a diagnostic test will be determined for patients with Lyme disease as compared with standard PCR and culture. The spirochetal burden in skin biopsy samples of erythema migrans (EM) skin lesions and spirochetal virulence factors in isolates from these lesions will be delineated, and the results will be correlated with clinical markers of spirochetal dissemination and severity of infection. Finally, cytokine and chemokine profiles in EM lesions from these patients will be determined and correlated with clinical markers of spirochetal dissemination and severity of infection and with blood test results of cellular and humoral immune function. These studies are likely to lead to more accurate tests for the serodiagnosis of Lyme disease, for direct detection of the spirochete, and for the assessment of spirochetal killing. In addition, greater knowledge of spirochetal virulence and host factors associated with severe disease may allow the development of laboratory markers for patients who would benefit from more intensive treatment.