TICK SALIVA INHIBITS INFLAMMATION IN MONOCYTES STIMULATED WITH B BURGDORFERI

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 这项研究的主要目的是通过检测在与伯氏疏螺旋体共培养过程中扁虱唾液对单核细胞、巨噬细胞和成纤维细胞等皮肤细胞相关免疫途径的基因组图谱以及随后与这些途径相关的蛋白质表达的影响，来检验这一假说。硬蜱的摄食过程延长，再加上细菌的传播，应该会在摄血部位产生强烈的炎症反应。然而，扁虱唾液中存在的因素可能会下调这种有害的反应。从伯氏杆菌刺激的人THP-1单核细胞中分离出转录本和/或上清液，并在有或没有壁虱唾液的情况下生长，用于人基因芯片、细胞因子珠阵列、ELISA和qRT-PCR实验。在与伯氏杆菌和唾液共培养的三个时间点收集的单核细胞RNA表明，唾液可以抑制某些促炎介质的表达，如IL-8和TLR-2，并对特定的分子如IL-10ra具有刺激作用，IL-10ra是已知的IL-10免疫抑制信号的介体。培养上清液通过抗原捕获酶联免疫吸附试验和细胞因子微球阵列分析炎性介质的产生。唾液治疗可显著降低THP-1单核细胞表达IL-6、IL-8和TNF-α等几种关键介质。用原代恒河猴成纤维细胞进行了类似的实验，我们的结果表明扁虱唾液对这些前哨皮肤细胞有相反的影响。唾液非但没有抑制，反而促进了某些促炎介质的产生，如IL-8和IL-6。

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.