ANTIBODY RESPONSES AGAINST MOG IN MARMOSET EAE

这项建议的长期目标是开发一种新的免疫治疗方法，用于在非人类灵长类动物中脱髓鞘EAE。为了实现这一长期目标，将追求以下具体目标：(1)鉴定雅克夏鱼的IgG1、VH和VL kappa谱系。利用基于聚合酶链式反应的方法，RACE和随后的DNA测序将可以鉴定出雅克希虫IgG1H和L链亚家族。(2)雅克逊对虾抗髓鞘蛋白抗体反应的多样性分析。通过构建一个组合F(Ab)文库，可以选择MOG和MBP的特异性克隆，并描述这些髓鞘蛋白在抗体反应中使用的多样性。(3)产生可溶性的MOG和MBP特异的F(Ab)片段，用于MOG免疫的日本血吸虫，以防止脱髓鞘。从与MOG或MBP高亲和力结合的噬菌体克隆中去除gIII蛋白，将允许表达可溶的F[ab]片段。高亲和力MOG特异性F[ab]将被测试其防止绒猴EAE脱髓鞘的能力。MBP特异性F[ab]将用作对照。如果这种免疫治疗方法被证明是成功的，那么基于类似原理的治疗方法可以为人类多发性硬化症开发。

The long-term objective of this proposal is to develop a new immunotherapeutic approach towards demyelinating EAE in a non- human primate. To achieve this long-term goal the following specific aims will be pursued: (1) Characterization the IgG1 VH and VL kappa repertoire in C. jacchus. Using the PCR based method RACE and subsequent DNA sequencing will permit the identification of C. jacchus IgG1 H and L chain subfamilies. (2) Analysis of the diversity of the antibody response against myelin proteins in C. jacchus. By constructing a combinatorial F(ab) library from C. jacchus IgG mRNA it will be possible to select MOG and MBP specific clones and to describe the diversity IgG1 VH and VL kappa usage in the antibody response to these myelin proteins. (3) Production of soluble MOG-and MBP-specific F(ab) fragments to be administered in MOG immunized C. jacchus to prevent demyelination. The removal of the gIII protein from the phage clones selected for high affinity binding to MOG or MBP, will permit the expression of soluble F[ab] fragments. High affinity MOG specific F[ab] will be tested in their ability to prevent demyelination in marmoset EAE. MBP specific F[ab] will be used as control. If this immunotherapeutic approach proves to be successful, therapies based on similar principles could be developed for human MS.