Development of an adenoviral vector system with adenovirus serotype 35 tropism; efficient transient gene transfer into primary malignant hematopoietic cells
Development of an adenoviral vector system with adenovirus serotype 35 tropism; efficient transient gene transfer into primary malignant hematopoietic cells
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DOI：
10.1002/jgm.543
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发表时间：
2004-06-01
影响因子：
3.5
通讯作者：
Fan, XL
中科院分区：
文献类型：
--
作者：
Nilsson, M;Ljungberg, J;Fan, XL
柯萨奇腺病毒受体（CAR）的缺乏阻碍了基于血清型5 （Ad5）的腺病毒载体介导的基因转移到恶性造血细胞。具有B类倾向的纤维重定向腺病毒载体可以潜在地绕过CAR的要求，促进有效的基因转移到恶性造血细胞中。方法利用嵌合纤维基因编码Ad5纤维尾部结构域、Ad35纤维轴和旋涡结构域，对多功能AdEasy系统进行了修饰，使其成为纤维重定向腺病毒载体。在具有Ad35纤维受体特异性的PGK启动子调控下，生成了基于ad5的绿色荧光蛋白（GFP）基因编码载体（Ad5F35-GFP）。将Ad5F35-GFP载体介导的基因转移效率与未经纤维修饰的Ad5-GFP载体进行比较，后者也在PGK启动子的控制下编码GFP基因。结果我们发现多种ad5难治性恶性髓系和B淋巴系对Ad5F35-GFP载体感染具有高度容忍度。重要的是，与Ad5-GFP载体相比，Ad5F35-GFP载体在原发性慢性髓性白血病（CML）细胞和慢性淋巴细胞白血病（CLL） B细胞的感染多重性（MOI）为100时具有优势。结论本研究将促进纤维重定向腺病毒载体的产生，并使原发性恶性造血细胞的瞬时遗传操作成为可能。版权所有：John Wiley Sons， Ltd. 2004
Background A paucity of coxsackie adenovirus receptor (CAR) hampers the adenovirus serotype 5 (Ad5)-based vector-mediated gene transfer into malignant hematopoietic cells. Fiber-retargeted adenoviral vectors with species B tropism can potentially bypass the CAR requirement and facilitate efficient gene transfer into malignant hematopoietic cells.Methods For feasible generation of fiber-retargeted adenoviral vectors, we have modified the versatile AdEasy system with a chimeric fiber gene encoding the Ad5 fiber tail domain and Ad35 fiber shaft and knob domains. An Ad5-based vector encoding the green fluorescent protein (GFP) gene under the control of the PGK promoter with Ad35 fiber receptor specificity was generated (Ad5F35-GFP). The Ad5F35-GFP vector-mediated gene transfer efficiency was compared with a fiber non-modified Ad5-GFP vector, which also encodes the GFP gene under the control of the PGK promoter.Results We demonstrated that a variety of Ad5-refractory malignant myeloid and B lymphoid cell lines were highly permissive to the Ad5F35-GFP vector infection. Importantly, primary chronic myeloid leukemic (CML) cells and chronic lymphocytic leukemia (CLL) B cells were superiorly transduced by the Ad5F35-GFP vector at a multiplicity of infection (MOI) of 100 compared with the Ad5-GFP vector.Conclusions Our study will facilitate the generation of fiber-retargeted adenoviral vectors and enable transient genetic manipulation of primary malignant hematopoietic cells. Copyright (C) 2004 John Wiley Sons, Ltd.