Studies of Immunity /Tolerance Relating to Factor VIII

与因子 VIII 相关的免疫/耐受性研究

• 批准号: 6680030
• 负责人: Basil Golding
• 金额: --
• 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/6680030
• 关键词: antigen antibody reaction; coagulation factor VIII; disease /disorder model; embryo /fetus; gene expression; hemophilia As; immune tolerance /unresponsiveness; immunity; immunogenetics; laboratory mouse; newborn animals; serology /serodiagnosis

Summary: The objective is to determine whether early exposure to F. VIII in the fetus or neonate abrogates antibody development in mouse hemophiliacs. B-domain-deleted F. VIII has been expressed in our laboratory in several vectors: adenovirus, adeno-associated virus and as DNA plasmids. All three have been shown to express functional F. VIII in vitro. In vivo experiments are underway to see if expression can be detected in serum of treated mice. Preliminary data show that the constructs can correct clotting times in hemophiliac mice. Future experiments will determine whether these constructs induce antibody responses in adult mice as expected. If antibodies are induced the constructs will be introduced earlier in the life of these mice to see whether they can be tolerized rather than immunized. First neonatal treatment will be tried and if this fails fetal treatment will be attempted to induce tolerance. In addition, this project will help decide which constructs are more effective in replacement therapy both in regard to levels of expression of F. VIII and in the ability of each construct to be immunogenic. Immune responses aginst F. VIII and against the vector will be assessed in terms of antibody and cellular responses. Antibodies and cytotoxic T cell responses against the vector can be expected to limit the usefulness of the particular vector.

总结：目的是确定早期接触F。胎儿或新生儿中的VIII消除了小鼠血友病患者的抗体形成。 B结构域缺失型F. VIII已经在我们的实验室中在几种载体中表达：腺病毒、腺相关病毒和作为DNA质粒。 这三个函数都可以表示函数F。VIII体外。 体内实验正在进行中，以观察是否可以在治疗小鼠的血清中检测到表达。 初步数据显示，该结构可以纠正血友病小鼠的凝血时间。 未来的实验将确定这些构建体是否如预期的那样在成年小鼠中诱导抗体应答。 如果抗体被诱导，则将在这些小鼠的生命早期引入构建体，以观察它们是否可以被耐受化而不是免疫。 将尝试第一次新生儿治疗，如果失败，将尝试胎儿治疗以诱导耐受。 此外，该项目将有助于决定哪些结构在替代治疗中更有效，无论是在F表达水平方面。VIII和每个构建体具有免疫原性的能力。 抗F.将根据抗体和细胞应答来评估VIII和针对载体的抗体和细胞应答。 可以预期针对载体的抗体和细胞毒性T细胞应答限制特定载体的有用性。