Tick feeding regulation by orthologous tick saliva serine protease inhibitors

描述(由申请人提供)：在美国，人类媒介传播的疾病主要是壁虱传播的。本研究的目的是鉴定、鉴定和验证美洲Amblyomma americanum(Amblyomma Americanum)和Xxodes Sigularis Sigularis Ticularis Sp(Serpins)作为白尾鹿抗硬蜱感染疫苗靶抗原的有效性。为了完成摄食，扁虱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、由蛇严格控制的途径。我们的实验室和其他人提出，在取食过程中，扁虱表达并向宿主体内注射蛇，以调节宿主对扁虱取食活动的逃避防御反应。在NIHR03项目的最后一年，我们已经筛选出10条美洲硬蜱唾液腺(SG)丝蛋白，并验证了其中2条蛇(S)、AamS6和AamS8分泌到硬蜱唾液中，这是它们在硬蜱饲养过程中注射到宿主中的代理。我们扩展了这些研究，并鉴定了14种对血食有反应的肩部扁虱和SG蛇类。在这项提案中，我们将利用NIHR03项目中开发的方案来验证14条我肩部SG蛇分泌到扁虱唾液中的有效性。我们观察到两种美洲硬蜱唾液蛇和一条蓖麻硬茧蜂唾液蛇在硬蜱中高度保守。因此，我们假设壁虱利用同源的壁虱唾液蛇来调节壁虱的摄食，免疫阻断壁虱唾液蛇的功能将阻止多个壁虱物种的饲养成功。白尾鹿(Odocoileus Virginianus)在美国主要人类硬虱传播疾病(TBD)、莱姆病、人类无浆体病、巴贝斯虫病和埃立克斯虫病以及南方硬蜱相关皮疹(STARI)的出现中具有重要作用。因此，该提案中的数据将与开发新技术以预防人类待命疾病的努力直接相关。这项申请有三个具体目标。第一个是验证16种血餐反应性的一号扁虱唾液腺毒蛇分泌到扁虱唾液中的情况。第二个研究是描述美洲硬蜱和肩隆本地硬蜱唾液蛇在硬蜱摄食调节中的作用(S)。目的：验证重组硬蜱唾液蛇毒抗原免疫白尾鹿后，白尾鹿对硬蜱的免疫力和记忆抗体反应性。

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.