Ruminating over host-parasite interaction models for fluke driven immune responses

反思侥幸驱动的免疫反应的宿主-寄生虫相互作用模型

• 批准号: 2878865
• 金额: --
• 依托单位: Aberystwyth University
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2878865
• 关键词: Ruminating; over; host; parasite; interaction

Rumen fluke (RF), Calicophoron daubneyi, are trematode parasites that infect ruminant livestock and given a changing climate, are increasing in prevalence. Heavy infections with immature RF can result in animal morbidity and mortality. Due to the relatively recent emergence of RF infections compared to other more pathogenic gastrointestinal helminths, there is limited knowledge as to how the parasite interacts with the host, especially in terms of the host immune response. It is well known that related pathogenic helminths, such as liver fluke, have the ability to modulate host immune responses to their benefit, allowing longer survival in the host. One important mechanism that has been demonstrated in several parasitic species is the use of extracellular vesicles (EVs), which have the ability to transport parasite-derived cargo molecules, which are bioactive, and cause immunomodulatory changes in the host. Adult RF are known to produce and release EVs (Huson et al., 2018), containing proteins previously identified as immune modulators in related species and also have the ability to modulate bacterial populations within the rumen (Allen et al. 2021). However, the mechanism used by these EVs to modulate the host tissue immune responses is unknown. In vitro models of animal diseases are desirable to develop an understanding of molecular host parasite interactions within a controlled experimental environment, allowing the reduction, refinement and replacement of animal studies. Cell culture models are commonly used in vitro, but do not account for multiple cell types within a tissue. As such, tissue explant models are a useful tool for providing ex vivo models. These explant models are also a novel method for helminth infections, but at present have not been utilised for helminth studies. To begin to elucidate the role of RF EVs on the host immune response, the present study aims to further establish new in vitro models that are relevant to RF predilection sites (ruminant gastrointestinal tract) and utilise established species-specific cell culture models to develop our understanding of RF immunomodulatory roles within the host. This project will complete key work to understand the host immune response to RF, which will be essential to understanding immune responses in co-infections with the more pathogenic liver fluke.The hypotheses to be tested in this studentship are: 1. EVs produced by adult RF will modulate the host immune response in rumen tissue explants and epithelial cells, as well as an immune specific cell line of bovine macrophages. 2. Specific predicted immune related proteins identified in EVs will produce immunomodulatory responses like that observed in whole EV studies. To investigate these hypotheses further, the following objectives will be completed: 1.Establish an in vitro explant tissue model of the bovine rumen for host-parasite interactions that can be maintained for a short term (up to 72 hours) successfully (e.g. tissue architecture and viability maintained) and be stimulated with EVs from adult RF to assess immune responses in the tissue via cytokine secretion and polyomics (transcriptome and proteome profiling). (Lead:AU)2.Verify results from objective 1 in an established model of rumen epithelial cells (Ji et al., 2021; using RT-PCR for targeted gene expression), as well as assessing cell viability and metabolism in response to EV stimulation. (Lead:QUB)3.Synthesise key immune modulator proteins constitutively expressed by adult RF EVs to test in the rumen tissue explant/epithelial cell model and assess immune responses to better understand mechanisms of action of bioactive molecules within EVs. (Lead:QUB on synthesis of proteins and AU on experimental work)4.Assess changes in immune parameters in bovine macrophages stimulated with EVs from adult RF over time to better understand immediate and longer-term immune responses. (Lead:AU)

瘤胃氟（RF），Calicophoron daubneyi是感染反刍动物牲畜并鉴于气候变化的巨大寄生虫。不成熟的RF的重型感染会导致动物的发病率和死亡率。由于与其他更为病的胃肠道舵相比，RF感染的出现相对较新，因此关于寄生虫如何与宿主相互作用的知识有限，尤其是在宿主免疫反应方面。众所周知，相关的致病性蠕虫（例如肝氟）具有调节宿主免疫反应对其受益的能力，从而使宿主的生存更长。在几种寄生虫物种中已证明的一种重要机制是使用细胞外囊泡（EV），这些囊泡具有运输寄生虫衍生的货物分子的能力，这些货物是生物活性的，并在宿主中引起免疫调节性变化。已知成年RF会产生和释放EV（Huson等，2018），其中包含先前被鉴定为相关物种中免疫调节剂的蛋白质，并且还具有调节瘤胃中细菌种群的能力（Allen等，2021）。但是，这些EV用于调节宿主组织免疫反应的机制尚不清楚。希望在受控的实验环境中对分子宿主寄生虫相互作用的了解，从而可以减少，改进和替代动物研究。细胞培养模型通常在体外使用，但不考虑组织中多种细胞类型。因此，组织外植体模型是提供离体模型的有用工具。这些外植体模型也是一种用于蠕虫感染的新方法，但目前尚未用于蠕虫研究。为了开始阐明RF电动汽车在宿主免疫反应中的作用，本研究旨在进一步建立与RF倾向位点有关的新的体外模型（反刍动物胃肠道），并利用已建立的物种特异性细胞培养模型来开发我们理解宿主内RF免疫调节作用。该项目将完成关键工作，以了解宿主对RF的免疫反应，这对于了解与更致病性肝氟的共同感染中的免疫反应至关重要。 RF将调节瘤胃组织外植体和上皮细胞中的宿主免疫反应，以及牛巨噬细胞的免疫特异性细胞系。 2。在电动汽车中鉴定出的特定预测的免疫相关蛋白将产生像整个EV研究中观察到的那样的免疫调节反应。为了进一步研究这些假设，将完成以下目标：1。建立一种体外植物瘤瘤瘤瘤的体外外植体组织模型，用于宿主 - 寄生虫相互作用，可以成功维持短期（长达72小时）（例如，组织结构）并维持生存能力），并通过成年RF的电动汽车刺激，以通过细胞因子的分泌和多合子（转录组和蛋白质组分析）评估组织中的免疫反应。 （铅：au）2。在瘤胃上皮细胞的既定模型中验证目标1的结果（Ji等，2021；使用RT-PCR用于靶向基因表达），并评估对EV的细胞生存能力和代谢。刺激。 （铅：Qub）3。与成人RF EV组成型表达的关键免疫调节剂蛋白在瘤胃组织外植体/上皮细胞模型中测试，并评估免疫反应，以更好地了解EV中生物活性分子的作用机制。 （铅：关于蛋白质和AU的合成Qub的实验性工作）4。随着时间的推移，成年RF刺激的牛巨噬细胞中免疫参数的变化，以更好地了解即时和长期的免疫反应。 （铅：au）