ROLE OF MONOCLONAL IGA IN TREATMENT OF CRYPTOSPORIDIOSIS

单克隆 IGA 在治疗隐孢子虫病中的作用

• 批准号: 3791866
• 负责人: SAUL TZIPORI
• 金额: --
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3791866
• 关键词: Cryptosporidium; antiprotozoal agents; cooperative study; cryptosporidiosis; disease /disorder model; host organism interaction; immunoglobulin A; immunotherapy; infant animal; laboratory mouse; monoclonal antibody; nonhuman therapy evaluation; protozoal antigen; severe combined immunodeficiency; surface antigens; tissue /cell culture

The research goals of Project 3 are to investigate strategies for the treatment of cryptosporidiosis based on the inhibition of parasite attachment and cell invasion by monoclonal IgAs directed against sporozoite surface epitopes. Attention will be focused on developing and characterizing monoclonal IgA antibodies against surface epitopes shared by exogenous (sporozoites) and endogenous (merozoites) infectious forms to maximize the efficiency of inhibition. Specific Aim 1 will be devoted to production and analysis of dimeric monoclonal IgA clones reactive against sporozoite and merozoite surface molecules. This work will be performed at Dr. Neutra's laboratory at the Children's Hospital, Harvard Medical School. Specific Aim 2 will demonstrate in animal models the ability of IgA clones to protect mice against Cryptosporidium infection. Hybridoma cells will be injected subcutaneously to produce IgA- secreting tumors (backpack). Cells will be injected to infant Balb/C mice, simultaneously challenged with Cryptosporidium oocysts to determine protection against infection, and to weaned SCID mice chronically infected to determine impact on clearing existing infection. The impact of an oral course of IgA antibodies will also be tested; monoclonal antibodies will be fed to infant mice simultaneously challenged with Cryptosporidium, and to weaned SCID mice chronically infected. A course of intravenous injections to SCID mice with evidence of hepato-biliary involvement will determine impact on Cryptosporidium infection of the biliary tree. This work will be performed jointly between Dr. Neutra's laboratory and Dr. Tzipori's at TUSVM, which are 30 miles apart. Specific Aim 3 will largely be performed at Dr. Flanigan's laboratory at Miriam Hospital, Brown University (50 miles distance). The nature of inhibition will be determined in cell culture system (HT29.74) developed by Dr. Flanigan. He will determine whether inhibition is caused by interference with, sporozoite binding, penetration, or development within the host cell. Project 3 is also responsible for program administration, and for providing services which include, i) production and regular supply of oocysts to all 3 projects, and ii) will be responsible for performing all animal challenge assays and analyses.

项目3的研究目标是研究策略 基于抑制的隐孢子虫病的治疗 单克隆 IgAs 引导的寄生虫附着和细胞侵袭 针对子孢子表面表位。 注意力将集中在 开发和表征单克隆 IgA 抗体 外源（子孢子）和内源共享的表面表位 （裂殖子）感染形式，以最大限度地提高效率 抑制。 具体目标 1 将致力于生产和分析 二聚体单克隆 IgA 克隆对子孢子有反应 裂殖子表面分子。 这项工作将在博士进行。 哈佛医学院儿童医院的 Neutra 实验室 学校。 具体目标2将在动物模型中证明IgA的能力 克隆以保护小鼠免受隐孢子虫感染。 杂交瘤细胞将被皮下注射以产生 IgA- 分泌肿瘤（背包）。 细胞将被注射给婴儿 Balb/C 小鼠，同时接受隐孢子虫卵囊攻击 确定对感染的保护作用，并使 SCID 小鼠断奶 慢性感染以确定对清除现有的影响 感染。 IgA 抗体口服疗程的影响将 也进行测试；单克隆抗体将被喂给幼年小鼠 同时用隐孢子虫进行攻击，并断奶 SCID 小鼠慢性感染。 静脉注射一个疗程 有肝胆系统受累证据的 SCID 小鼠将 确定对胆管树隐孢子虫感染的影响。 这项工作将由 Neutra 博士的实验室联合进行 和 TUSVM 的 Tzipori 博士的诊所相距 30 英里。 具体目标 3 将主要在 Flanigan 博士的诊所进行 布朗大学米里亚姆医院实验室（50 英里 距离）。 抑制的性质将在细胞中决定 Flanigan 博士开发的培养系统（HT29.74）。 他会 确定抑制是否是由干扰引起的， 子孢子在宿主内结合、渗透或发育 细胞。 项目 3 还负责项目管理，并负责 提供的服务包括：i) 生产和定期供应 向所有 3 个项目提供卵囊，并且 ii) 将负责 进行所有动物挑战试验和分析。