TICK SALIVA INHIBITS INFLAMMATION IN MONOCYTES STIMULATED WITH B BURGDORFERI

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 本研究的主要目的是通过检查蜱唾液在与B共培养期间对人单核细胞的体外影响，检验蜱唾液在伯氏疏螺旋体背景下对与皮肤细胞（如单核细胞/巨噬细胞和成纤维细胞）相关的免疫途径的基因组谱以及对与这些途径相关的后续蛋白质表达的广泛影响这一假设。burgdorferi。硬蜱的长时间进食过程与细菌传播相结合，应导致血液进食部位的强烈炎症反应。然而，蜱唾液中存在的因素可能会下调这种有害的反应。 从用B刺激的人THP-1单核细胞分离的转录物和/或上清液。在人微阵列、细胞因子珠阵列、ELISA和qRT-PCR实验中使用了在存在或不存在蜱唾液的情况下生长的伯氏螺旋体。从与B共培养期间的三个时间点收获单核细胞RNA。Burgdorferi和唾液的研究表明，唾液可以对某些促炎介质如IL-8和TLR-2的表达具有抑制作用，以及对特定分子如IL-10 ra（IL-10的免疫抑制信号的已知介质）具有刺激作用。通过抗原捕获ELISA和细胞因子珠阵列分析上清液的炎症介质产生。用唾液处理显著降低了THP-1单核细胞的几种关键介质（包括IL-6、IL-8和TNF-α）的表达。 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。而不是抑制，唾液增强某些促炎介质，如IL-8和IL-6的生产。

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.