ANTIBODY RESPONSES AGAINST MOG IN MARMOSET EAE

本建议的长期目标是开发一种新的免疫治疗方法来治疗非人灵长类动物的EAE脱髓鞘。为了实现这一长期目标，将追求以下具体目标：(1)表征紫蝽IgG1 VH和VL kappa曲目。利用基于RACE的PCR方法和随后的DNA测序将允许鉴定紫茎草IgG1 H和L链亚家族。(2)刺槐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，将有可能选择MOG和MBP特异性克隆，并描述IgG1 VH和VL kappa在抗体对这些髓鞘蛋白的反应中的多样性。(3)可溶性MOG和mbp特异性F（ab）片段的产生，用于MOG免疫的紫针菇预防脱髓鞘。从选择与MOG或MBP高亲和力结合的噬菌体克隆中去除gIII蛋白，将允许可溶性F[ab]片段的表达。将测试高亲和力MOG特异性F[ab]在狨猴EAE中防止脱髓鞘的能力。MBP特异性F[ab]作为对照。如果这种免疫治疗方法被证明是成功的，基于类似原理的治疗方法可以用于人类多发性硬化症。

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.