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的研究目标是研究以下策略 基于对隐孢子虫病抑制作用的治疗 单抗免疫球蛋白诱导的寄生虫附着和细胞侵袭 抗子孢子表面表位。我们将把注意力集中在 抗人免疫球蛋白A单抗的研制和鉴定 外源(子孢子)和内源共有的表面表位 (裂殖子)感染形式，以最大限度地提高 抑制力。 具体目标1将致力于生产和分析 抗子孢子的二聚体单抗IgA克隆和 裂殖子表面分子。这项工作将在Dr。 哈佛大学医学院儿童医院的中性实验室 学校。 特定目标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。 杂交瘤细胞将被注射到皮下产生IgA- 分泌性肿瘤(背包)。细胞将被注射给婴儿 BALB/C小鼠，同时攻击隐孢子虫卵囊 确定对感染的保护作用，并断奶的SCID小鼠 慢性感染，以确定对清理现有 感染。口服一疗程的IgA抗体的影响将 也要进行检测；单抗将喂给幼鼠 同时攻击隐孢子虫和断奶SCID 慢性感染的小鼠。一个疗程的静脉注射以治疗 有肝胆受累证据的SCID小鼠将 确定隐孢子虫对胆道感染的影响。 这项工作将由中性子博士的实验室联合完成 齐波里博士在相距30英里的德克萨斯大学支持向量机工作。 具体目标3将主要在弗拉尼根博士的 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确定 培养系统(HT29.74)由弗拉尼根博士开发。他会的 确定抑制是否由干扰引起， 子孢子在宿主内结合、穿透或发育的子孢子 手机。 项目3还负责项目管理，并负责 提供服务，包括：一)生产和定期供应 向所有3个项目提供卵囊，以及ii)将负责 执行所有动物挑战测试和分析。