The protease network that regulates innate immunity in mosquitoes

项目摘要 蚊子的先天免疫系统是其媒介能力的关键决定因素。这包括 支持属中原生动物寄生虫种的发育和传播的能力 按蚊是世界范围内人类疟疾的主要传播媒介。洞察 先天免疫效应机制的调节仍然不完全，但对我们的免疫系统至关重要。 对这一最重要的人类媒介传播的宿主-病原体相互作用的基本了解 疾病长期的目标是了解免疫系统的调节。冈比亚将通报最新情况 以及未来的病媒控制战略。本申请的目的是在全球范围内确定 通过确定细胞外蛋白酶网络内的相互作用来调节免疫系统， 在不同微生物感染的情况下激活调理作用并将其与黑化联系起来。的理由 拟议研究是关于调节蚊子免疫力的蛋白酶网络的详细信息 可用于预测使用微生物的新型病媒控制策略的长期效力， 药物，并操纵感染结果。根据我们的初步数据，以下三个具体目标 本论文的主要工作如下：（1）确定蛋白酶及其同源物对蚊子的作用 免疫力;（2）评估蛋白酶网络对免疫力和蚊子适应性的影响;以及（3） 使用网络科学可视化蚊子的免疫调节网络。在第一个目标下，我们将 测试clip-丝氨酸蛋白酶及其同系物形成功能模块的假设， 通过定义它们的切割模式，遗传相互作用和精确的免疫反应， 生化功能在第二个目标下，蛋白酶网络对病原体的潜在影响 将使用常见的微生物挑战来评估抗性和耐受性以及蚊子适应性 模型和生命表分析。在第三个目标下，标准的网络科学方法将用于 将系统中的所有蛋白酶相互作用可视化为静态多层网络，并分析该网络 以推断蛋白水解流通过连接调理作用和黑化，并确定关键分子 控制着免疫力这项研究具有创新性，因为它将首次评估蛋白酶 级联作为一个单一的，综合的网络，控制蚊子的体液免疫在不同的免疫 挑战此外，该项目将使用网络科学作为一种高度创新的方法来研究 蚊子的先天免疫，如果成功，将是昆虫免疫学领域的变革。这 该项目是重要的，因为它将提供全面的了解蛋白酶的贡献， 这一系统对蚊子健康的影响以及该系统在克服感染方面的局限性。最终这 知识可以用来操纵感染的结果，从而告知新的发展， 疾病控制战略，旨在破坏疟疾寄生虫在其载体中的发展。

PROJECT SUMMARY The innate immune system of mosquitoes is a critical determinant of their vector competence. This includes the ability to support development and transmission of the protozoan parasite species in the genus Plasmodium by Anopheles mosquitoes, the principal vectors of human malaria world-wide. Insight into the regulation of innate immune effector mechanisms remains incomplete, but is vitally important to our fundamental understanding of host-pathogen interactions in this most important human vector-borne disease. The long-term goal is to understand immune system regulation in An. gambiae to inform current and future vector control strategies. The objective of this application is to globally identify mechanisms of immune system regulation by determining the interactions within the extracellular protease network that activate and link opsonization to melanization in the context of distinct microbial infections. The rationale for the proposed research is that detailed information on the protease network that regulate mosquito immunity could be employed to predict long-term efficacy of novel vector control strategies that employ microbial agents, and manipulate infection outcome. Guided by our preliminary data, the following three specific aims will be pursued: (1) Determine the interactions of proteases and their homologs that are critical for mosquito immunity; (2) Assess the impact of the protease network on immunity and mosquito fitness; and (3) Visualize the immunoregulatory network in mosquitoes using network science. Under the first aim, we will test the hypothesis that clip-serine proteinases and their homologs form functional modules that are required for optimal immune responses by defining their cleavage patterns, genetic interactions, and precise biochemi-cal function. Under the second aim, the potential effect of the protease network on pathogen resistance and tolerance as well as mosquito fitness will be assessed using common microbial challenge models and life table analyses. Under the third aim standard network science approaches will be used to visualize all protease interactions in the system as a static multilayered network and to analyze this network to infer proteolytic flow through that links opsonization and melanization and to identify the key molecules that control immunity. The proposed research is innovative, as it will for the first time evaluate protease cascades as a single, integrated network that controls mosquito humoral immunity during diverse immune challenges. Additionally, this project will use network science as a highly innovative approach to the study of mosquito innate immunity, which if successful will be transformative to the field of insect immunology. This project is significant as it will provide comprehensive understanding of the contribution of the protease network to mosquito health as well as the limitations of the system in overcoming infection. Ultimately, this knowledge could be employed to manipulate infection outcome and thus inform the development of new disease control strategies that aim at disrupting malaria parasite development in its vector.