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.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 我们的最初目标是检验以下假设：滴答唾液会对皮肤细胞（如血液单核细胞和成纤维细胞）在暴露于螺旋体Borrelia burgdorferi的促炎性介体的产生/转录中产生抑制或抑制作用。进行了以下体外实验，以检验我们的假设：（1）在存在和不存在tick唾液的情况下，从THP-1细胞系的激活人单核细胞共培养； （2）在存在和不存在tick唾液的情况下，人类和恒河生成纤维细胞共同培养。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。在微阵列实验中使用了从THP-1单核细胞中提取的RNA。通过独立的实时PCR实验获得了微阵列结果的验证。我们的结果表明，当前的范式可能是对宿主，载体和病原体之间更复杂的相互作用的过度简化。 tick唾液不仅能够抑制，而且还可以增强B. burgdorferi在单核细胞中引起的炎症介质产生和转录。在恒河类和人类成纤维细胞中，即使在没有螺旋体的情况下，它也能够引起促炎性介质的产生。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长度，并且可以从早期抑制性转变为晚期增强，反之亦然。