Braun S E, Aronovich E L, Anderson R A, Crotty P L, McIvor R S, Whitley C B
Department of Genetics and Cell Biology, University of Minnesota, Minneapolis 55455.
Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11830-4. doi: 10.1073/pnas.90.24.11830.
To explore the possibility of using gene transfer to provide iduronate-2-sulfatase (IDS; EC 3.1.6.13) enzyme activity for treatment of Hunter syndrome, an amphotropic retroviral vector, L2SN, containing the human IDS coding sequence was constructed and studied for gene expression in vitro. Lymphoblastoid cell lines (LCLs) from patients with Hunter syndrome were transduced with L2SN and expressed high levels of IDS enzyme activity, 10- to 70-fold higher than normal human peripheral blood leukocytes or LCLs. Such L2SN-transduced LCLs failed to show accumulation of 35SO4 into glycosaminoglycan (35SO4-GAG), indicating that recombinant IDS enzyme participated in GAG metabolism. Coculture of L2SN-transduced LCLs with fibroblasts from patients with Hunter syndrome reduced the accumulation of 35SO4-GAG. These results demonstrated retroviral-mediated IDS gene transfer into lymphoid cells and the ability of such cells to provide recombinant enzyme for intercellular metabolic cross-correction.
为了探索利用基因转移来提供艾杜糖醛酸-2-硫酸酯酶（IDS；EC 3.1.6.13）的酶活性以治疗亨特综合征的可能性，构建了一种含有人类IDS编码序列的双嗜性逆转录病毒载体L2SN，并对其在体外的基因表达进行了研究。用L2SN转导亨特综合征患者的淋巴母细胞系（LCLs），这些细胞系表达高水平的IDS酶活性，比正常人外周血白细胞或LCLs高10至70倍。这种经L2SN转导的LCLs未能显示出35SO4积累到糖胺聚糖（35SO4-GAG）中，这表明重组IDS酶参与了GAG代谢。将经L2SN转导的LCLs与亨特综合征患者的成纤维细胞共培养可减少35SO4-GAG的积累。这些结果证明了逆转录病毒介导的IDS基因转移到淋巴细胞中，以及这些细胞为细胞间代谢交叉校正提供重组酶的能力。
