TICK SALIVA INHIBITS INFLAMMATION IN MONOCYTES STIMULATED WITH B BURGDORFERI

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 我们最初的目标是验证这样一种假设，即当接触伯氏疏螺旋体时，扁虱唾液对血液单核细胞和成纤维细胞等真皮细胞释放的促炎介质的产生/转录具有单独的抑制或抑制作用。为了验证我们的假设，我们进行了以下体外实验：(1)将THP-1细胞系中激活的人单核细胞与伯氏杆菌在存在和不存在壁虱唾液的情况下共培养；(2)将人和恒河猴成纤维细胞与伯氏杆菌在存在和不存在壁虱唾液的情况下进行共培养。每个实验的细胞培养上清液用于细胞因子ELISA法和/或多重细胞因子微球阵列。共培养3个时间点从THP-1单核细胞中提取RNA用于基因芯片实验。通过独立的实时荧光聚合酶链式反应实验对芯片结果进行验证。我们的结果表明，目前的模式可能是对宿主、病媒和病原体之间更复杂的相互作用的过度简化。壁虱唾液不仅能抑制和增强伯氏杆菌诱导的单核细胞炎症介质的产生和转录。在恒河猴和人类成纤维细胞中，即使在没有螺旋体的情况下，它也能够诱导促炎介质的产生。此外，在某些情况下，唾液对单核细胞的影响取决于接触唾液的时间长短，并可能从早期抑制转换到后期增强，反之亦然。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Our initial goal was to test the hypothesis that tick saliva would have solely suppressive or inhibitory effects on the production/transcription of pro-inflammatory mediators put out by dermal cells such as blood monocytes and fibroblasts when exposed to the spirochete Borrelia burgdorferi. The following in vitro experiments were conducted in order to test our hypothesis: (1) co-culture of activated human monocytes from the THP-1 cell line with B. burgdorferi in the presence and absence of tick saliva; and (2) co-culture of human and rhesus fibroblasts with B. burgdorferi in the presence and absence of tick saliva. Cell culture supernatants from each experiment were used in cytokine ELISA and/or multiplex cytokine bead arrays. RNA extracted from THP-1 monocytes at three time points during co-culture was used in microarray experiments. Validation of microarray results was gained by independent real-time PCR experiments. Our results suggest that the current paradigm may be an oversimplification of a more complex interplay between host, vector and pathogen. Tick saliva was able not only to inhibit but also enhance inflammatory mediator production and transcription elicited by B. burgdorferi in monocytes. In rhesus and human fibroblasts it was able to elicit production of pro-inflammatory mediators even in the absence of the spirochete. Moreover, saliva effects on monocytes in some cases depended on the length of exposure to the saliva, and could switch from early inhibitory to late enhancing, and vice-versa.