Serine Proteinase Inhibitors (Serpins) in Tick Feeding and Physiology

丝氨酸蛋白酶抑制剂（Serpin）在蜱饲喂和生理学中的作用

• 批准号: 7673631
• 负责人: ALBERT MULENGA
• 金额: $ 7.28万
• 依托单位: TEXAS A&M UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-15 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7673631
• 关键词: Amblyomma; Antibodies; Arthropod Vectors; Blood coagulation; Containment; Data; Data Analyses; Development; Disease; Family; Future; Gene Silencing; Goals; Host Defense; Human; Immunoglobulin Variable Region; Inflammation; Knowledge; Lead; Life; Messenger RNA; Methods; Mutation; Names; Organism; Pathway interactions; Peptides; Physiology; Positioning Attribute; Protease Inhibitor; Protein Family; Proteins; Protocols documentation; Public Health; Publishing; RNA Interference; Regulation; Saliva; Salivary Glands; Serine Proteinase Inhibitors; Serpins; Technology; Testing; Tick Control; Ticks; United States; Vector-transmitted infectious disease; Western Blotting; acaricide; acronyms; base; feeding; pathogen; polyclonal antibody; prevent; programs; public health relevance

DESCRIPTION (provided by applicant): In the United States (US) ticks transmit more vector borne disease agents than any other vector arthropod. Limitations associated with current acaricide based tick control strategies that threaten the future sustainability of tick borne illnesses containment programs, have necessitated the need for development of alternative tick control strategies. Our long-term goals are to identify important tick proteins, which if blocked, can compromise the tick's ability to feed, acquire and transmit disease agents. One important such class of proteins, are serine proteinase inhibitors that are commonly referred to by its acronym "serpin", which is the largest class of proteinase inhibitors that have been found in most forms of life. In humans, serpins are involved in regulation of important of proteolytic pathways that are essential to life such as blood coagulation with serpin malfunctions arising from natural mutations causing numerous debilitating diseases. We have recently described a family of 19 Amblyomma americanum serpins that we have named as "Lospins", an acronym for Lone Star tick serpins. Sequence and expression data analyses have revealed that 10 of the 19 Lospin cDNAs encode for Lospin proteins that are potentially secreted into tick saliva during tick feeding. Our hypothesis is that, the 10 Lospins are secreted into the host during tick feeding to facilitate the tick's escape from host defense and that disrupting their function will prevent the tick from feeding. In this proposal we will characterize six of the 10 Lospins via two specific aims. The first is to validate secretion into tick saliva of six selected Lospin proteins. The rationale is that Lospins detected in tick saliva will represent candidates that are secreted into the host during tick feeding and will be selected for further study. We will raise specific polyclonal antibodies to candidate Lospins and use them in standard western blotting analyses of collected tick saliva. The second, we will be to suppress expression of Lospins selected in aim 1 and study the effects on tick feeding. The rationale is that suppression of an essential Lospin will prevent the tick from feeding and it will be selected for future study. A. americanum is an important public health pest that is commonly encountered tick species particularly in southern and Atlantic states. The immediate utility of results from this proposal will be advancing our knowledge on the physiology of this important pest. This data will position us to rationale new methods to control this tick. PUBLIC HEALTH RELEVANCE: In the United States ticks transmit more vector borne disease agents than any other vector arthropod. Limitations associated with current acaricide based tick control strategies that threaten the future sustainability of containment programs for tick borne illnesses, have necessitated the need for development of alternative tick control strategies. Identification of important tick proteins that regulate tick physiology and facilitate tick feeding is important before alternative tick control methods can be developed.

描述（由申请方提供）：在美国（US），蜱传播的病媒病原体多于任何其他病媒节肢动物。与当前基于杀螨剂的蜱虫控制策略相关的局限性威胁到蜱虫传播疾病遏制计划的未来可持续性，因此需要开发替代蜱虫控制策略。我们的长期目标是确定重要的蜱蛋白，如果被阻断，可能会损害蜱的进食，获得和传播疾病的能力。一类重要的此类蛋白质是丝氨酸蛋白酶抑制剂，其通常以其首字母缩写“丝氨酸蛋白酶抑制剂”来指代，其是已在大多数生命形式中发现的最大类别的蛋白酶抑制剂。在人类中，丝氨酸蛋白酶抑制剂参与调节对生命至关重要的重要蛋白水解途径，例如血液凝固，其中丝氨酸蛋白酶抑制剂功能障碍由导致许多衰弱性疾病的天然突变引起。我们最近描述了一个家庭的19 Amblyomma americanum蛇针，我们已经命名为“Lospins”，一个首字母缩写为孤独的星星蜱蛇针。序列和表达数据分析显示，19个Lospin cDNA中有10个编码Lospin蛋白，这些蛋白在蜱进食期间可能分泌到蜱唾液中。我们的假设是，10个Lospin在蜱虫进食过程中分泌到宿主体内，以促进蜱虫逃离宿主防御，破坏它们的功能将阻止蜱虫进食。在本提案中，我们将通过两个具体目标来描述10个Lospins中的6个。第一个是验证六种选定的Lospin蛋白分泌到蜱唾液中。基本原理是，在蜱唾液中检测到的Lospins将代表在蜱进食期间分泌到宿主中的候选物，并将被选择用于进一步研究。我们将提出候选Lospins的特异性多克隆抗体，并将其用于收集的蜱唾液的标准蛋白质印迹分析。第二，我们将抑制目标1中选择的Lospins的表达，并研究对蜱摄食的影响。基本原理是抑制必需的Lospin将阻止蜱虫进食，并将其选择用于未来研究。A.美洲蜱是一种重要的公共卫生害虫，特别是在南部和大西洋各州，通常会遇到蜱类。这项建议的结果的直接效用将推进我们对这一重要害虫的生理学知识。这些数据将使我们能够合理地采用新的方法来控制这种蜱虫。 公共卫生相关性：在美国，蜱虫传播的媒介传播疾病比任何其他媒介节肢动物都多。与当前基于杀螨剂的蜱虫控制策略相关的限制威胁到蜱虫传播疾病遏制计划的未来可持续性，因此需要开发替代蜱虫控制策略。在开发替代蜱控制方法之前，鉴定调节蜱生理学和促进蜱进食的重要蜱蛋白是重要的。