Gut Epithelial Cell Receptors for C parvum Invasion

描述（由申请人提供）：细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。目前治疗这种寄生虫感染的治疗策略主要集中在破坏孢子体附着和侵入肠上皮细胞的过程上。这些研究已经确定了两种孢子体配体gp900和gp40/15，它们粘附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，但生产功能性重组小孢子虫配体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。缺乏功能性重组小弧菌配体阻碍了寄生虫入侵细胞所用上皮细胞受体的鉴定。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：具体目的1：生成重组gp900糖蛋白，并表征天然gp900和重组gp900在体外与肠上皮细胞的结合。特异性目的2：鉴定gp900和gp40/15的肠上皮细胞受体，这些实验结果将为小隐孢子虫孢子体附着和侵袭过程提供更深入的了解，并将促进隐孢子虫病治疗策略的制定。该奖项将为候选人O'Connor博士提供在分子水平上研究小孢子虫配体与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项目提供基础。奥康纳博士将由她的赞助人沃德博士提供建议，沃德博士是胃肠病学家，也是公认的肠道寄生虫专家。她的共同导师Kopin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，并为年轻科学家提供了特殊的环境。

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.