National Scientific Center "Institute of Experimental and Clinical Veterinary Medicine", Kharkiv, Ukraine.
Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, Ceske Budejovice, Czechia.
Front Cell Infect Microbiol. 2023 Jan 20;13:1112952. doi: 10.3389/fcimb.2023.1112952. eCollection 2023.
We developed a new simple method to assess the composition of proteinaceous components in the saliva of , the main vehicle for pathogen transmission and a likely source of bioactive molecules acting at the tick-vertebrate host interface. To collect naturally expectorated saliva from the ticks we employed an artificial membrane feeding technique using a simple, chemically defined diet containing phagostimulants and submitted native saliva samples collected in this way for liquid chromatography-mass spectrometry (LC-MS) analysis. These experiments were conducted with groups of uninfected ticks as well as with infected with . The ticks exhibited a fair feeding response to the tested diet with engorgement rates reaching as high as 60-100% of ticks per feeding chamber. The LC-MS analysis identified a total of 17 and 15 proteins in saliva samples from the uninfected and infected nymphs, respectively. Importantly, the analysis was sensitive enough to detect up to 9 different proteins in the samples of saliva containing diet upon which as few as 6 nymphal ticks fed during the experiments. Some of the proteins recognized in the analysis are well known for their immunomodulatory activity in a vertebrate host, whereas others are primarily thought of as structural or "housekeeping" proteins and their finding in the naturally expectorated tick saliva confirms that they can be secreted and might serve some functions at the tick-host interface. Most notably, some of the proteins that have long been suspected for their importance in the vector-pathogen interactions of spirochetes were detected only in the samples from infected ticks, suggesting that their expression was altered by the persistent colonization of the tick's salivary glands by spirochetes. The simple method described herein is an important addition to the toolbox available to study the vector-host-pathogen interactions in the rapidly feeding soft ticks.
我们开发了一种新的简单方法来评估 的蛋白质成分的组成， 是病原体传播的主要载体，也是作用于蜱-脊椎动物宿主界面的生物活性分子的可能来源。为了从蜱中收集自然吐出的唾液，我们采用了一种简单的人工膜喂养技术，使用含有取食刺激物的化学定义饮食，并对以这种方式收集的天然唾液样本进行液相色谱-质谱（LC-MS）分析。这些实验是在未感染的蜱组以及感染 的蜱组中进行的。 蜱对测试饮食表现出相当大的取食反应，饱食率高达每个喂食室的 60-100%的蜱。LC-MS 分析鉴定了来自未感染和感染 的若虫唾液样本中的 17 种和 15 种蛋白质，分别。重要的是，该分析足够灵敏，能够在含有饮食的唾液样本中检测到多达 9 种不同的蛋白质，而在实验期间，每个喂食室中只有 6 只若虫蜱进食了这些饮食。在分析中识别出的一些蛋白质以其在脊椎动物宿主中的免疫调节活性而闻名，而另一些则主要被认为是结构或“管家”蛋白，它们在自然吐出的蜱唾液中的发现证实了它们可以被分泌出来，并可能在蜱-宿主界面发挥某些功能。最值得注意的是，一些长期以来被怀疑在 螺旋体的媒介-病原体相互作用中具有重要意义的蛋白质仅在感染蜱的样本中被检测到，这表明它们的表达被螺旋体对蜱唾液腺的持续定植所改变。本文所述的简单方法是研究快速进食软蜱中的媒介-宿主-病原体相互作用的工具包的重要补充。
