TICK SALIVA INHIBITS INFLAMMATION IN MONOCYTES STIMULATED WITH B BURGDORFERI

蜱唾液抑制布氏 B 氏菌刺激的单核细胞炎症

• 批准号: 8172987
• 负责人: MARIO TOMAS PHILIPP
• 金额: $ 6.18万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8172987
• 关键词: Antigens; Black-legged Tick; Blood; Borrelia burgdorferi; Cells; Coculture Techniques; Computer Retrieval of Information on Scientific Projects Database; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Funding; Genomics; Goals; Grant; Harvest; Human; IL8 gene; Immune; Immunosuppressive Agents; In Vitro; Inflammation; Inflammation Mediators; Inflammatory Response; Institution; Interleukin-10; Interleukin-6; Lead; Macaca mulatta; Mediator of activation protein; Pathway interactions; Process; Production; RNA; Research; Research Personnel; Resources; Saliva; Sentinel; Signal Transduction; Site; Skin; Source; Testing; Ticks; Time; Transcript; Tumor Necrosis Factor-alpha; United States National Institutes of Health; cytokine; feeding; human TNF protein; macrophage; monocyte; protein expression; research study; response; transmission process

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The primary goal of this study was to test the hypothesis that tick saliva, in the context of Borrelia burgdorferi, can have widespread effects on both the genomic profile of immune pathways associated with skin cells such as monocytes/macrophages and fibroblasts and on the subsequent protein expression associated with those pathways by examining the in vitro effects of tick saliva on human monocytes during their co-culture with B. burgdorferi. The prolonged feeding process of ixodid ticks, in combination with bacterial transmission, should lead to a robust inflammatory response at the blood-feeding site. Yet, factors present in tick saliva may down-regulate such deleterious responses. Transcripts and/or supernatants isolated from human THP-1 monocytes stimulated with B. burgdorferi and grown in the presence or absence of tick saliva were used in human microarray, cytokine bead array, ELISA, and qRT-PCR experiments. Monocyte RNA harvested from three time points during co-culture with B. burgdorferi and saliva illustrated that saliva can have a suppressive effect on the expression of certain pro-inflammatory mediators, such as IL-8 and TLR-2 as well as have a stimulatory effect on specific molecules such as IL-10ra, a known mediator of the immunosuppressive signal of IL-10. Supernatants were analyzed via antigen-capture ELISA and cytokine bead array for inflammatory mediator production. Treatment with saliva significantly reduced the expression of several key mediators including IL-6, IL-8 and TNF-alpha by THP-1 monocytes. Similar experiments were conducted using primary rhesus fibroblasts and our results indicate that tick saliva has an opposite effect on these sentinel skin cells. Rather than inhibiting, saliva enhanced production of certain pro-inflammatory mediators, such as IL-8 and IL-6.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 本研究的主要目标是检验以下假设：在伯氏疏螺旋体的背景下，蜱唾液可以对与单核细胞/巨噬细胞和成纤维细胞等皮肤细胞相关的免疫途径的基因组谱以及随后的蛋白质产生广泛影响通过检查蜱唾液在与伯氏疏螺旋体共培养期间对人类单核细胞的体外影响，研究与这些途径相关的表达。蜱蜱的长期摄食过程与细菌传播相结合，会导致吸血部位出现强烈的炎症反应。然而，蜱唾液中存在的因素可能会下调这种有害反应。 从用伯氏疏螺旋体刺激并在存在或不存在蜱唾液的情况下生长的人THP-1单核细胞分离的转录物和/或上清液用于人微阵列、细胞因子珠阵列、ELISA和qRT-PCR实验。与伯氏疏螺旋体和唾液共培养期间从三个时间点收获的单核细胞 RNA 表明唾液可以对某些促炎介质（例如 IL-8 和 TLR-2）的表达具有抑制作用，并且具有刺激作用对特定分子（如 IL-10ra）的影响，IL-10ra 是 IL-10 免疫抑制信号的已知介质。通过抗原捕获 ELISA 和细胞因子珠阵列分析上清液以了解炎症介质的产生。唾液处理显着降低了 THP-1 单核细胞的几种关键介质的表达，包括 IL-6、IL-8 和 TNF-α。 使用原代恒河猴成纤维细胞进行了类似的实验，我们的结果表明蜱唾液对这些前哨皮肤细胞具有相反的作用。唾液非但没有抑制，反而增强了某些促炎介质的产生，例如 IL-8 和 IL-6。