Antigen-mediated growth control of hybridoma cells via a human artificial chromosome
Antigen-mediated growth control of hybridoma cells via a human artificial chromosome
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DOI：
10.1016/j.bbagen.2006.10.014
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发表时间：
2007-02-01
影响因子：
3
通讯作者：
Nagamune, Teruyuki
中科院分区：
文献类型：
--
作者：
Kawahara, Masahiro;Inoue, Toshiaki;Nagamune, Teruyuki
人类人工染色体（HAC）载体具有足以满足基因治疗载体要求的几个特征，包括稳定的附加型维持和长期转基因表达的介导。在这项研究中，我们采用了一种抗原介导的遗传修饰细胞扩增（AMEGA）系统，采用抗体/细胞因子受体嵌合体，触发生长信号响应于同源无毒抗原，并将其应用于生长控制的TAC转移细胞，通过添加一种抗原，不同的细胞因子，可能表现出多效性效应。我们以前构建了一种新的HAC载体，21 Δ qHAC，来自人类21号染色体，安置在CHO细胞中。在这里，我们构建了一个HAC载体携带ScFv-gp 130嵌合体响应荧光素结合的BSA（BSA-FL）以及模型转基因，增强型绿色荧光蛋白（EGFP），在CHO细胞。通过微细胞介导的染色体转移将修饰的HAC转移到白细胞介素（IL）-6依赖的杂交瘤细胞7 TD 1中，随后发现细胞在不存在IL-6的情况下显示出BSA-FL依赖的细胞生长和持续的EGFP表达。AMEGA系统与HAC技术的组合将有助于通过赋予遗传修饰细胞生长优势来增加基因治疗的功效。(c)2006 Elsevier B. V.保留所有权利。
Human artificial chromosome (HAC) vectors possess several characteristics sufficient for the requirements of gene therapy vectors, including stable episomal maintenance and mediation of long-term transgene expression. In this study, we adopted an antigen-mediated genetically modified cell amplification (AMEGA) system employing an antibody/cytokine receptor chimera that triggers a growth signal in response to a cognate nontoxic antigen, and applied it to growth control of HAC-transferred cells by adding an antigen that differed from cytokines that may manifest pleiotropic effects. We previously constructed a novel HAC vector, 21 Delta qHAC, derived from human chromosome 2 1, housed in CHO cells. Here, we constructed an HAC vector harboring an ScFv-gp130 chimera responsive to fluorescein-conjugated BSA (BSA-FL) as well as a model transgene, enhanced green fluorescent protein (EGFP), in CHO cells. The modified HAC was transferred into interleukin (IL)-6-dependent hybridoma 7TD1 cells by microcell-mediated chromosome transfer, and the cells were subsequently found to show BSA-FL-dependent cell growth and sustained expression of EGFP in the absence of IL-6. The AMEGA system in combination with HAC technology will be useful for increasing the efficacy of gene therapy by conferring a growth advantage on the genetically modified cells. (c) 2006 Elsevier B.V. All rights reserved.