The protease network that regulates innate immunity in mosquitoes

项目总结 蚊子的先天免疫系统是它们媒介能力的关键决定因素。这包括 支持该属原生动物寄生虫物种发育和传播的能力 世界范围内人类疟疾的主要媒介--按蚊传播的疟原虫。洞察 对先天免疫效应机制的调节仍然不完整，但对我们的 对这种最重要的人类媒介传播的宿主和病原体相互作用的基本了解 疾病。长期目标是了解免疫系统在体内的调节。冈比亚向海流通报 以及未来的媒介控制策略。此应用程序的目标是在全球范围内识别 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 在不同的微生物感染的背景下，激活并将调理作用与黑化联系起来。其基本原理是 这项拟议的研究是关于调节蚊子免疫的蛋白酶网络的详细信息 可以用来预测使用微生物的新型媒介控制策略的长期效果 毒剂，并操纵感染结果。根据我们的初步数据，以下三个具体目标 将继续进行：(1)确定对蚊子至关重要的蛋白酶及其同系物的相互作用 (2)评估酶网络对免疫和蚊子健康的影响；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 测试以下假设：片段丝氨酸蛋白酶及其同系物形成的功能模块是 通过定义它们的切割模式、基因相互作用和精确的 生化功能。在第二个目标下，蛋白酶网络对病原菌的潜在影响 将使用常见的微生物挑战来评估抗药性和耐受性以及蚊子的适合性 模型和生命表分析。在第三个目标下，将使用标准的网络科学方法来 将系统中的所有蛋白酶相互作用可视化为一个静态的多层网络，并分析该网络 推断蛋白水解流通过该环节的调理和黑化，并确定关键分子 控制着豁免权。这项拟议的研究是创新的，因为它将首次评估蛋白酶 作为一个单一的、集成的网络级联，在不同的免疫过程中控制蚊子的体液免疫 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 蚊子的天然免疫，如果成功，将对昆虫免疫学领域产生革命性的影响。这 该项目意义重大，因为它将提供对蛋白酶的贡献的全面了解 委员会还注意到该网络对蚊子健康的影响，以及该系统在克服感染方面的局限性。归根结底，这 可以利用知识来操纵感染结果，从而为开发新的 疾病控制战略，旨在破坏疟疾寄生虫在其媒介中的发展。

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.