Molecular Aspects of Tsetse and Trypanosome Transmission

该应用程序用于预防非洲锥虫病，这是非洲最被忽视的疾病之一 由采采赛传播的寄生非洲锥虫引起。缺乏有效的工具来遏制 哺乳动物的感染和动物宿主的存在需要病媒控制来抗击疾病。 我们将研究采采集-锥虫相互作用对传播动力学的影响。用于传输到 发生时，锥虫首先在中肠(MG)感染，然后移动到苍蝇的口器，以 进入并定植唾液腺(SG)。消除大多数寄生虫的主要障碍是 蝇类发生在MG。我们已经证明了一种寄生虫介导的媒介生理的操纵过程 在感染早期暂时性地降低中肠屏障的完整性，使寄生虫能够绕过周围的 矩阵(PM)屏障。这种操控过程的核心是哺乳动物寄生虫表面蛋白，变种 表面糖蛋白(VSG)，在感染过程的早期进入肠腔，干扰 Tsetse的PM合成通过microRNA(miR-275)。通过操纵性的 再次处理使感染寄生虫的MG重新进入管腔以定植SG。我们将使用 要调查的跨学科研究计划： 1.降低PM疗效的机制和寄生虫VSG蛋白的不同成分是 在感染过程的早期对这种干扰负责。我们还将调查寄生虫 在感染过程的后期实现PM减少的组件，因为寄生虫从MG迁移到SGS以 变速箱。我们将进行媒介和寄生虫转录图谱分析，以发现潜在的介体。 生物体内的对话。 2.Tsetse microRNA(MiR275)通过识别其下游分子靶点在PM合成中的作用 通过转录组和RiBoseq图谱，并通过使用双荧光素酶分析在 S2细胞株，免疫共沉淀法。 3.采采虫-寄生虫在野外自然感染中的相互作用以验证寄生虫-病媒的对话 在实验室中观察，并确定PM修改对建立联合 感染多种寄生虫物种和品系。利用野外苍蝇，我们将确定 评估PM屏障的流行病学意义的寄生虫传播过程。 总的来说，我们的研究将提供关于适应性和操纵性过程的基本知识 影响重要媒介的媒介能力和疾病传播，并将揭示潜在的目标 用于通过传播阻断策略或副转基因应用进行干扰，以减少疾病。

This application is on prevention of African Trypanosomiasis, one of the most neglected diseases of Africa caused by parasitic African trypanosomes transmitted by tsetse. The absence of effective tools to curb infections in the mammal and the presence of animal reservoirs necessitate vector control to combat disease. We will investigate tsetse-trypanosome interactions that influence transmission dynamics. For transmission to occur, trypanosomes first establish infections in the midgut (MG) and then move to the fly's mouthparts to access and colonize the salivary glands (SG). The major barrier that eliminates parasites from the majority of flies occurs in the MG. We have shown that a parasite mediated manipulative process of vector's physiology transiently reduces midgut barrier integrity early in the infection to enable the parasites to bypass the peritrophic matrix (PM) barrier. At the core of this manipulative process is the mammalian parasite surface proteins, Variant Surface Glycoproteins (VSGs), shed into the gut lumen early in the infection process, which interfere with tsetse's PM synthesis acting through a microRNA (miR-275). Loss of PM integrity through a manipulative process again enables MG infecting parasites to re-enter into the lumen to colonize SG. We will use an interdisciplinary research plan to investigate: 1. The mechanisms that reduce PM efficacy and the different components of the parasite VSG protein that are responsible for this interference early in the infection process. We will also investigate the parasite components that enable PM reduction later in the infection process as parasites migrate from MG to SGs for transmission. We will perform vector and parasite transcriptomic profiling to discover potential mediators of the intra-organismal dialogue. 2. The role of the tsetse microRNA (miR275) in PM synthesis by identifying its downstream molecular targets through transcriptome and Riboseq profiling and by validating these targets using a dual-luciferase assay in a S2 cell line and through co-immunoprecipation assays. 3. Tsetse-parasite interactions in natural infections in the field to validate the parasite-vector dialogue we observe in the laboratory, and to determine the influence of PM modification on establishment of co- infections with multiple parasite species and strains. Using field flies, we will determine the course of parasite transmission processes to assess the epidemiological significance of PM barriers. Collectively, our studies will provide fundamental knowledge on adaptive and manipulative processes that influence vector competence and disease transmission in an important vector and will reveal potential targets for interference by transmission blocking strategies or paratransgenic applications to reduce disease.