Molecular Studies of Immunoparasitology in Snails

描述（由申请方提供）：淡水蜗牛光滑双脐螺在人类血吸虫曼氏血吸虫的传播中起着不可或缺的作用。增加我们对这种蜗牛内部防御的基本机制的了解，可以帮助开发控制血吸虫病的新方法，血吸虫病仍然感染着全球2亿人。关于螺旋体和它们的蜗牛宿主之间的分子相互作用，以及B之间的分子相互作用，还有很多未知之处。glabrata - S. mansoni模型系统是迄今为止评估抗性和相容性的潜在机制的最佳方法。这项研究是建立在以前的研究基础上的，这些研究表明蜗牛血蛋白（称为FREPs（纤维蛋白原相关蛋白质））在抗线粒体防御中的功能作用。他们进一步受到我们的发现的启发，这些发现表明B。glabrata通过点突变和重组产生了令人惊讶的FREP基因多样性，这一发现沿着其他实验室的工作，引发了对无脊椎动物产生多样化防御反应能力的普遍重新思考。在本申请中，已广泛修订，以响应以前的审查，我们开发了一种通用的方法来评估功能的推定免疫效应（FREPs是主要的，但不是唯一的，效应器可用于测试使用这种方法）在影响的能力S。mansoni在B.光滑的在这样做时，重要的是从它们参与抵抗感染的角度以及作为相容性的一般决定因素来检查候选效应分子。具体而言，我们将进一步开发RNA干扰（RNAi）和重组蛋白/抗体的方法来检查FREPs的功能作用，影响寄生虫在不相容的主机，使用在体内和体外的方法。我们最近确定的其他候选人，即革兰氏阴性菌结合蛋白（GNBP）和肽聚糖识别蛋白（PGRP），控制上游Toll/Imd途径的关键分子的蜗牛同源物，也可用于研究，从而进一步拓宽了我们对蜗牛免疫的理解。最后，我们将尝试破坏S. mansoni实现这种中断的能力将是一个重要的进步，因为兼容性确保了血吸虫病在流行地区的持续传播。这项研究将使我们能够获得有价值的见解的功能FREPs和其他候选人，甚至更重要的是，开发一个系统的普遍适用性，用于评估任何蜗牛候选分子的作用，在影响结果之间的遭遇S。曼索尼和它的蜗牛宿主

DESCRIPTION (provided by applicant): The freshwater snail Biomphalaria glabrata plays an indispensible role in transmission of the human blood fluke, Schistosoma mansoni. Increasing our understanding of the fundamental mechanisms of internal defense of this snail could help develop novel methods of control of schistosomiasis which still infects 200 million people world-wide. Much remains unknown regarding the molecular interplay between schistosomes and their snail hosts, and the B. glabrata - S. mansoni model system is by far the best for assessing underlying mechanisms of resistance and compatibility. The proposed investigations are built on previous studies suggestive of a functional role of snail blood proteins called FREPs (fibrinogen-related proteins) in anti-schistosome defenses. They are further inspired by our discoveries showing that B. glabrata generates a surprising diversity of FREP genes through point mutation and recombination, a discovery that along with work in other labs has provoked a general reconsideration of the capacity of invertebrates to generate diversified defense responses. In this application, which has been extensively revised in response to the previous review, we develop a general approach for assessing the functionality of putative immune effectors (FREPs are the main, but not the sole, effectors available to be tested using this approach) in influencing the ability of S. mansoni to develop successfully in B. glabrata. In so doing, it is important to examine candidate effector molecules both from the perspective of their involvement in resistance to infection, and as general determinants of compatibility. Specifically, we will further develop RNA interference (RNAi) and recombinant protein/antibody approaches to examine the functional role of FREPs in affecting the development of parasites in incompatible hosts, using both in vivo and in vitro approaches. Other candidates we have recently identified, namely gram-negative bacteria binding protein (GNBP) and peptidoglycan recognition proteins (PGRPs), the snail homologs of key molecules controlling upstream Toll/Imd pathways, are also available for study, thus further broadening our understanding of snail immunity. Finally, we will attempt to disrupt the successful parasitization of compatible snails by S. mansoni. The ability to effect such disruption would be an important advance because compatibility ensures the continued transmission of schistosomiasis in endemic areas. This study will enable us to gain valuable insights into the functionality of FREPs and other candidates, and even more importantly, to develop a system of general applicability for evaluating the role of any snail candidate molecules in influencing the outcome of an encounter between S. mansoni and its snail host.