EFFICIENT CDNA EXPRESSION VECTORS FOR STABLE AND TRANSIENT EXPRESSION OF HLA-DR IN TRANSFECTED FIBROBLAST AND LYMPHOID-CELLS
EFFICIENT CDNA EXPRESSION VECTORS FOR STABLE AND TRANSIENT EXPRESSION OF HLA-DR IN TRANSFECTED FIBROBLAST AND LYMPHOID-CELLS
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DOI：
10.1016/0198-8859(91)90092-n
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发表时间：
1991-08-01
期刊：
影响因子：
2.7
通讯作者：
JARAQUEMADA, D
中科院分区：
文献类型：
--
作者：
LONG, EO;ROSENBRONSON, S;JARAQUEMADA, D
构建了具有几个有用特征的cDNA表达载体。首先，劳斯肉瘤病毒的长末端重复序列被用作启动子，以在人和小鼠来源的各种细胞中获得高水平的表达。第二，顺式连接的表达单位，赋予耐霉酚酸或新霉素类似物G418插入，以促进表达感兴趣的cDNA的转染细胞的分离。第三，通过在猿COS细胞中复制，这些载体可用于有效的瞬时表达。将编码人11类主要组织相容性复合物抗原的DR-α或DR-β链的cDNA片段插入这些载体中，共转染小鼠和人成纤维细胞后获得高水平的细胞表面HLA-DR抗原。这些载体还成功地用于纠正来自先天性免疫缺陷患者的II类阴性B细胞系不能将肽抗原呈递给DR限制性T细胞。
cDNA expression vectors with several useful features were constructed. First, the long terminal repeat of Rous sarcoma virus was used as a promoter to obtain high levels of expression in various cells of human and mouse origin. Second, cis-linked expression units that confer resistance either to mycophenolic acid or the neomycin analog G418 were inserted to facilitate the isolation of transfected cells expressing the cDNA of interest. Third, by replicating in simian COS cells, these vectors can be used for efficient transient expression. cDNA fragments encoding the DR-alpha or DR-beta-chains of human class 11 major histocompatibility complex antigens were inserted into these vectors and high levels of cell surface HLA-DR antigen were obtained after cotransfection into mouse and human fibroblasts. These vectors were also successfully used to correct the inability of a class II-negative B cell line, derived from a patient with a congenital immunodeficiency, to present peptide antigen to DR-restricted T cells.