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: 8358087
• 负责人: MARIO TOMAS PHILIPP
• 金额: $ 3.72万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8358087
• 关键词: Borrelia burgdorferi; Cell Culture Techniques; Cell Line; Cells; Coculture Techniques; Complex; Dermal; Enzyme-Linked Immunosorbent Assay; Exposure to; Fibroblasts; Funding; Genetic Transcription; Goals; Grant; Human; In Vitro; Inflammation; Inflammation Mediators; Length; Macaca mulatta; National Center for Research Resources; Order Spirochaetales; Primates; Principal Investigator; Production; RNA; Research; Research Infrastructure; Resources; Saliva; Source; Testing; Ticks; Time; United States National Institutes of Health; Validation; cost; cytokine; monocyte; pathogen; research study; vector

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.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们最初的目标是测试这样的假设，即当暴露于螺旋体伯氏疏螺旋体时，蜱唾液将对由皮肤细胞（如血液单核细胞和成纤维细胞）产生的促炎介质的产生/转录具有单独的抑制或抑制作用。为了验证我们的假设，进行了以下体外实验：（1）来自THP-1细胞系的活化的人单核细胞与B的共培养。在存在和不存在蜱唾液的情况下的burgdorferi;和（2）人和恒河猴成纤维细胞与B的共培养。在存在和不存在蜱唾液的情况下的伯氏螺旋体。将来自每个实验的细胞培养上清液用于细胞因子ELISA和/或多重细胞因子珠阵列。在共培养期间的三个时间点从THP-1单核细胞中提取的RNA用于微阵列实验。通过独立的实时PCR实验验证芯片结果。我们的研究结果表明，目前的范式可能是一个更复杂的宿主，病媒和病原体之间的相互作用过于简单化。 蜱唾液不仅能抑制而且能增强B诱导的炎症介质的产生和转录。单核细胞中的Burgdorferi。在恒河猴和人成纤维细胞中，即使在没有螺旋体的情况下，它也能够引起促炎介质的产生。此外，在某些情况下，唾液对单核细胞的影响取决于暴露于唾液的时间长度，并且可以从早期抑制转变为晚期增强，反之亦然。