Real-time Imaging Analysis of Vector-borne Lyme Borreliosis Pathogenesis & Persis

媒介传播莱姆疏螺旋体病发病机制的实时成像分析

• 批准号: 8424969
• 负责人: Linda K. Bockenstedt
• 金额: $ 42.85万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8424969
• 关键词: Acute; Antibiotic Therapy; Antibiotics; Antibodies; Antigenic Variation; Back; Blood; Borrelia burgdorferi; Ceftriaxone; Cells; Chronic; Chronic Phase; Clinical; Complement; Connective Tissue; Disease; Doxycycline; Electrons; Etiology; Exhibits; Fc Receptor; Future; Goals; Host Defense; Human; Image; Image Analysis; Imaging Techniques; Immune; Immune response; Immunocompetent; In Vitro; Individual; Infection; Ingestion; Ixodes; Learning; Leptospira interrogans; Life; Ligand Binding; Ligands; Lipoproteins; Lyme Disease; Lymphatic; Mammals; Mediating; Membrane; Membrane Proteins; Microscopy; Movement; Mus; North America; Order Spirochaetales; Organism; OspC protein; Pathogenesis; Pattern; Phagocytes; Phagocytosis; Phase; Population; Process; Property; Proteins; Public Health; Relative (related person); Research Personnel; Resistance; Role; Site; Skin; Sorting - Cell Movement; Speed; Staging; Stimulus; Surface; Symptoms; System; Techniques; Tendon structure; Testing; Ticks; Time; Time Study; Tissues; Toll-like receptors; Variant; Vector-transmitted infectious disease; Video Microscopy; Xenodiagnosis; cell motility; experience; feeding; fluidity; genetic manipulation; in vivo; insight; killings; macrophage; molecular imaging; monocyte; mouse model; mutant; outer surface lipoprotein; public health relevance; tick borne spirochete; transmission process; uptake; vector

DESCRIPTION (provided by applicant): Lyme disease, due to infection with the Ixodes tick-borne spirochete Borrelia burgdorferi (Bb) is the most common vector-borne disease in North America and a significant public health concern. Although usually amenable to antibiotic therapy, Bb infection can be followed by prolonged clinical symptoms, especially if treatment is delayed. Recent studies in mice reveal that under some circumstances, spirochetes can persist in connective tissue for extended periods after antibiotic treatment. Central to resolving the clinical issue of spirochete persistence after antibiotic therapy is an understanding of how Bb evades the normal host defenses that should complement antibiotic effects. Our preliminary findings of Bb interactions with phagocytes in vitro suggest that Bb outer surface membrane proteins (Osps) are highly mobile and that this feature, along with spirochete motility, facilitate its escape from near total engulfment by phagocytes, even after opsonization. We further show the feasibility of using multiphoton imaging to obtain time-resolved images of tick feeding and Bb movements within the murine host. This application will use in vitro imaging techniques and in vivo real-time multiphoton microscopy to further evaluate the contribution of Bb Osp mobility and spirochete motility to phagocyte evasion and Bb persistence within the mammalian host. Using a panel of mutant/transformant Bb with differential survival in mice, we will evaluate through confocal, electron and videomicroscopy the membrane mobility of their dominantly expressed Osp and the capacity of the Bb variants to escape engulfment by human monocytes and murine macrophages. We will then use real-time multiphoton imaging of live anesthesized mice to examine the requirement for OspC in tick-transmitted Bb infection and dissemination from the skin inoculation site, and effects of OspC deficiency on spirochete motility in vivo. Finally, we will use multiphoton microscopy to examine the changes in Bb motility patterns over time as it adapts to persist within mice, and the effects of antibiotics on Bb when administered during acute and chronic phases of infection. These studies will provide insight into the role of membrane protein mobility and spirochete motility in Bb evasion of immune destruction in vivo, and insight into how Bb may persist after antibiotic therapy for Lyme disease. Moreover, our results will also provide a powerful, well-characterized system for future real-time studies of the effects of specific genetic manipulations of the tick vector, the spirochete, and/or the mammal on tick-borne Bb infection.

描述（由申请人提供）：莱姆病，由于感染了ixodes tick-borne spirochete Borrelia burgdorferi（BB）是北美最常见的媒介传播疾病，并且是一个重大的公共卫生问题。尽管通常可以接受抗生素疗法，但BB感染可以长时间临床症状，尤其是在治疗延迟的情况下。在小鼠中的最新研究表明，在某些情况下，螺旋体可以在抗生素治疗后长时间在结缔组织中持续存在。在抗生素治疗后解决螺旋体持久性临床问题的核心是了解BB如何逃避应补充抗生素作用的正常宿主防御措施。我们在体外与吞噬细胞相互作用的初步发现表明，BB外表面膜蛋白（OSPS）也非常流动，即使在打击后，该特征以及螺旋体运动性也可以促进其从近乎完全吞噬吞噬细胞吞噬的逃脱。我们进一步显示了使用多光子成像获得鼠宿主内滴答喂养和BB运动的时间分辨图像的可行性。该应用将使用体外成像技术和体内实时多光子显微镜，以进一步评估BB OSP迁移率和螺旋体运动对哺乳动物宿主内吞噬细胞逃避和BB持久性的贡献。我们将使用一组突变体/转化的BB在小鼠中具有差异存活率，我们将通过共焦，电子和视频显微镜评估其主要表达的OSP的膜迁移率以及BB变体的能力逃避人类单核细胞和小鼠巨噬细胞的能力。然后，我们将使用现场麻醉小鼠的实时多光子成像来检查tick传输的BB感染中OSPC的需求，并从皮肤接种部位传播，以及OSPC缺乏对体内螺旋体运动的影响。最后，我们将使用多光子显微镜检查BB运动模式随时间的变化，因为它适应了小鼠的持续存在，以及在感染的急性和慢性相期间给予抗生素对BB的影响。这些研究将洞悉膜蛋白迁移率和螺旋体运动在BB逃避体内免疫破坏中的作用，并洞悉BB抗生素治疗莱姆病后如何持续存在。此外，我们的结果还将为未来的实时研究提供一个强大的，良好的系统，以实时研究tick载体的特定遗传操作，螺旋体和/或哺乳动物对tick传播BB感染的影响。