Gut Epithelial Cell Receptors for C parvum Invasion

描述(申请人提供)：微小隐孢子虫是一种胃肠道寄生虫，在免疫功能低下的宿主中引起慢性、严重和经常致命的腹泻疾病，目前尚无治疗方法。目前开发治疗这种寄生虫感染的治疗方法的策略主要集中在破坏子孢子附着和侵入肠道上皮细胞的过程。这些研究已经确定了两个子孢子配体gp900和gp40/15，它们附着在上皮细胞上，并介导附着和侵袭。编码这些糖蛋白的基因已经被鉴定、克隆和表达，但功能重组的微小弧菌配体的生产需要在真核系统中表达，以产生对上皮细胞结合至关重要的特定碳水化合物表位。缺乏功能性重组微小弧菌配体阻碍了对这种寄生虫用来入侵细胞的上皮细胞受体的识别。本项目的主要目的是分离和鉴定gp900和gp40/15的上皮细胞受体。本项目的具体目的是：1：产生重组gp900糖蛋白，并鉴定天然和重组gp900与体外培养的肠上皮细胞的结合。特殊目的2：鉴定肠上皮细胞gp900和gp40/15的受体。这些实验结果将为更深入地了解微小隐孢子虫子孢子的附着和侵袭过程提供更多的信息，并将推动开发治疗隐孢子虫病的策略。这一奖项将为候选人奥康纳博士提供在分子水平上研究微小弧菌配体-上皮细胞受体相互作用的机会，并为她在宿主-寄生虫相互作用领域建立独立的研究计划奠定基础。奥康纳的赞助人沃德博士将为奥康纳提供建议。沃德博士是一位胃肠病专家，也是公认的肠道寄生虫专家。她的另一位导师科平博士在识别和表征胃肠道受体方面拥有丰富的经验。新英格兰医学中心与塔夫茨大学和更广泛的波士顿科学界合作，为研究人员提供了多样化和优秀的科学资源，对于年轻科学家来说是一个特殊的环境。

DESCRIPTION (provided by applicant):Cryptosporidium parvum is a gastrointestinal parasite that causes chronic, severe, and frequently fatal diarrheal disease in immunocompromised hosts, for which there is no treatment. Current strategies for the development of therapies to treat this parasitic infection have focused on disruption of the processes of sporozoite attachment and invasion into intestinal epithelial cells. These studies have identified two sporozoite ligands, gp900 and gp40/15, that adhere to epithelial cells and mediate attachment and invasion. The genes encoding these glycoproteins have been identified, cloned, and expressed, but production of functional recombinant C. parvum ligands requires expression in eukaryotic systems to generate the particular carbohydrate epitopes critical for epithelial cell binding. The lack of functional recombinant C. parvum ligands has hampered identification of the epithelial cell receptors used by the parasite to invade cells. The major goal of this project is to isolate and identify the epithelial cell receptors for gp900 and gp40/15. The specific aims of this project are:Specific Aim 1: To generate recombinant gp900 glycoproteins and characterize binding of native and recombinant gp900 to intestinal epithelial cells in vitro.Specific Aim 2: To identify intestinal epithelial cell receptors for gp900 and gp40/15Results from these experiments will provide greater insight into the processes of C. parvum sporozoite attachment and invasion and will advance the goal of developing treatment strategies for cryptosporidiosis. This award will provide the candidate, Dr. O'Connor, with the opportunity to study C. parvum ligand-epithelial cell receptor interactions at the molecular level, and provide the groundwork for her to establish an independent research program in the field of host-parasite interactions. Dr. O'Connor will be advised by her sponsor, Dr. Ward, a gastroenterologist and a recognized expert on enteric parasites. Her co-mentor, Dr. Kopin, has extensive experience identifying and characterizing gastrointestinal receptors. New England Medical Center, in association with Tufts University and the wider Boston scientific community provides researchers with diverse and excellent scientific resources, and is exceptional environment for a young scientist.