Preclinical studies of lymphocyte gene therapy for mild Hunter syndrome (mucopolysaccharidosis type II)
Preclinical studies of lymphocyte gene therapy for mild Hunter syndrome (mucopolysaccharidosis type II)
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DOI：
10.1089/hum.1996.7.3-283
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发表时间：
1996-02-10
影响因子：
4.2
通讯作者：
Whitley, CB
中科院分区：
文献类型：
--
作者：
Braun, SE;Pan, D;Whitley, CB
为了探索离体淋巴细胞基因治疗轻度亨特氏综合征（粘多糖样变性II型）的可行性，我们评估了逆转录病毒介导的艾杜糖醛酸-2-硫酸酯酶（IDS）编码序列的基因转移到外周血淋巴细胞从酶缺陷的个人（PBL（MPS））。通过在长末端重复序列（LTR）（在载体L2 SN中）或巨细胞病毒（CMV）早期启动子（载体LNC 2）的转录调控下插入IDS cDNA来构建莫洛尼鼠白血病病毒衍生的逆转录病毒载体。高滴度的病毒生产细胞产生嗜酸性PA 317包装细胞。在用抗CD 3抗体和白细胞介素-2（IL-2）体外刺激T淋巴细胞3天后，在接下来的3天中的每一天转导PBL（MPS）一次。7 - 21天后，评价培养的PBL（MPS）的基因转移和IDS比活性。尽管基因转移效率为5%或更低，但L2 SN转导的PBL（MPS）的异质群体具有高水平的IDS酶活性（456 U/mg/hr +/- SD 292）。
To explore the feasibility of ex vivo lymphocyte gene therapy for mild Hunter syndrome (mucopolysaccharidosis type II), we evaluated retrovirus-mediated gene transfer of the iduronate-2-sulfatase (IDS) coding sequence into peripheral blood lymphocytes from enzyme-deficient individuals (PBL(MPS)). Moloney murine leukemia virus-derived retroviral vectors were constructed by inserting the IDS cDNA under transcriptional regulation of the long terminal repeat (LTR) (in vector L2SN) or the cytomegalovirus (CMV) early promoter (vector LNC2). High-titer virus-producer cells were generated using amphotropic PA317 packaging cells. After 3 days of in vitro stimulation of T lymphocytes with anti-CD3 antibody and interleukin-2 (IL-2), PBL(MPS) were transduced once on each of the next 3 days. Seven to 21 days later, cultured PBL(MPS) were evaluated for gene transfer and IDS specific activity. Heterogeneous populations of L2SN-transduced PBL(MPS) had high levels of IDS enzyme activity (456 U/mg per hr +/- SD 292) despite a gene transfer efficiency of 5% or less. Owing to overexpression of IDS in that percentage of PBL(MPS) successfully transduced, IDS activity was increased above the deficiency found in patients with Hunter syndrome (