摘要
为研究皮肤特异性高效表达hCTLA4-Ig分子对移植皮肤存活及受体免疫功能的影响,充分实现hCTLA4-Ig分子在皮肤移植中的免疫调控功能,利用K14(角蛋白14)基因启动子,构建了hCTLA4-Ig分子皮肤组织特异性表达载体,并通过受精卵原核显微注射技术制备了K14/hCTLA4-Ig转基因小鼠并建立了品系。通过RT-PCR和Northernblot分析表明,hCTLA4-Ig在转基因小鼠体内呈皮肤组织特异性高效表达;以GAPDH基因的表达量为内部参照分析表明,hCTLA4-Ig的表达水平在不同世代之间以及转基因个体的不同生命时相点之间保持相对恒定,说明皮肤组织特异性稳定表达hCTLA4-Ig的转基因小鼠品系已被建立。
hCTLA4-Ig,which is capable of inhibiting T cell activation and rendering T cell to be anergy by blocking co-stimulatory signal pathway, is recognized as a potential therapeutic molecule to extend the survival of skin graft in burn medicine. To investigate whether the survival of skin graft can be further extended by skin-specific expression of hCTLA4-Ig, a skin-specific expression vector was constructed by pacing the encoding sequence of hCTLA4-Ig under the drive of K14 promoter,and a transgenic mouse line was established with the vector. RT-PCR and Northern blot assay indicated that hCTLA4-Ig was skin-specifically expressed in transgenic mice at a rather high level. Compared with the expression of GAPDH, the expression of hCTLA4-Ig remained constant through generations and over lifetime. The results of this paper indicated that a transgenic mouse line skin-specifically and constitutively expressing hCTLA4-Ig had been established.
基金
国家"十五"重点科技攻关项目(编号:2001BA710B)资助~~
