Tick feeding regulation by orthologous tick saliva serine protease inhibitors

描述（由申请人提供）：在美国，人媒传播疾病主要由蜱传播。本研究的目的是鉴定、表征和验证美洲钝眼蜱和肩突硬蜱蜱唾液丝氨酸蛋白酶抑制剂（serpins）作为白色尾鹿抗蜱感染免疫的疫苗靶抗原。为了完成进食，蜱虫必须克服宿主防御反应、血液凝固、补体激活和炎症，所有这些都是丝氨酸蛋白酶介导的途径，这些途径由丝氨酸蛋白酶抑制剂严格控制。我们的实验室和其他人提出，在进食过程中，蜱表达并注射丝氨酸蛋白酶抑制剂到宿主中，以调节宿主对蜱进食活动的防御反应的逃避。在一个正在进行的NIHR03项目的最后一年，我们筛选了10个A。美洲蜱（Aam）的唾液腺（SG）丝氨酸蛋白酶抑制剂，并验证了其中2种丝氨酸蛋白酶抑制剂（S），AamS6和AamS8分泌到蜱唾液中，这代表它们在蜱进食期间注射到宿主中。我们已经扩展了这些研究，并确定了14个血餐反应I。肩胛蜱SG丝氨酸蛋白酶抑制剂。在本提案中，我们将利用NIHR03项目中开发的方案来验证14 I的分泌。肩胛肌SG丝氨酸蛋白酶抑制剂进入蜱唾液。我们观察到，两个A。americanum蜱唾液丝氨酸蛋白酶以及I.蓖麻蜱唾液丝氨酸蛋白酶在蜱中高度保守。因此，我们假设，蜱利用orthopathic蜱唾液丝氨酸蛋白酶抑制剂来调节蜱的喂养和免疫阻断蜱唾液丝氨酸蛋白酶抑制剂的功能，将防止喂养成功的多个蜱种。白色尾鹿（Odocoileus virginianus）在美国主要人类蜱传疾病（TBD）、莱姆病、人类无形体病、巴贝虫病和埃里希体病以及南方蜱相关皮疹疾病（STARI）的出现中的重要性已被认识到。因此，本提案中的数据将与开发预防人类TBD的新技术的努力直接相关。本申请有三个具体目标。第一是验证16血餐反应性I的分泌。肩胛肌蜱唾液腺丝氨酸蛋白酶抑制剂进入蜱唾液。第二个是描述A的角色。americanum和I.肩胛骨天然蜱唾液丝氨酸蛋白酶抑制剂在蜱摄食调节中的作用。验证用重组蜱唾液丝氨酸蛋白酶抑制剂抗原免疫的白色尾鹿对蜱侵染的蜱免疫和回忆性抗体应答。

DESCRIPTION (provided by applicant): In the United States human vector borne diseases are primarily tick borne. The goals of this research are identification, characterization and validation of Amblyomma americanum and Ixodes scapularis tick saliva serine protease inhibitors (serpins) as vaccine target antigens for immunization of white tailed deer against tick infestation. To complete feeding, ticks must overcome host defense reactions, blood coagulation, complement activation and inflammation, all of which are serine protease mediated pathways that are tightly controlled by serpins. Our lab and others have proposed that during feeding, ticks express and inject serpins into the host to regulate evasion of host defense response to tick feeding activity. In an ongoing NIHR03 project in its final year we have screened 10 A. americanum (Aam) tick salivary gland (SG) serpin, and validated that 2 of the serpins (S), AamS6 and AamS8 are secreted into tick saliva, which is proxy for their injection into the host during tick feeding. We have extended these studies and identified 14 blood meal responsive I. scapularis tick SG serpins. In this proposal we will utilize protocols developed in the NIHR03 project to validate secretion of the 14 I. scapularis SG serpins into tick saliva. We have observed that the two A. americanum tick saliva serpins as well as the I. ricinus tick saliva serpin are highly conserved in ticks. Thus, we hypothesize that ticks utilize orthologous tick saliva serpins to regulate tick feeding and that immunologically blocking tick saliva serpin functions will prevent feeding success of multiple tick species. The importance of white tailed deer (Odocoileus virginianus) in the emergence of major human tick borne diseases (TBD) in the United States, Lyme disease, human anaplasmosis, babesiosis and ehrlichiosis and the southern tick-associated rash illness (STARI) has been recognized. Thus data in this proposal will be directly relevant towards efforts to develop novel technologies to prevent human TBD. There are 3 specific aims in this application. The first is to validate secretion of 16 blood meal responsive I. scapularis tick salivary gland serpins into tick saliva. The second is to characterize role(s) of A. americanum and I. scapularis native tick saliva serpins in tick feeding regulation. To validate tick immunity and anamnestic antibody response to tick infestation of white tailed deer immunized with recombinant tick saliva serpin antigens.