Efficient gene transfer into human trophoblast cells with adenovirus vector containing chimeric type 5 and 35 fiber protein.
Efficient gene transfer into human trophoblast cells with adenovirus vector containing chimeric type 5 and 35 fiber protein.
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DOI：
10.1248/bpb.27.2046
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发表时间：
2004-12
影响因子：
2
通讯作者：
N. Koizumi;H. Mizuguchi;M. Kondoh;M. Fujii;T. Hayakawa;Yoshiteru Watanabe
中科院分区：
文献类型：
--
作者：
N. Koizumi;H. Mizuguchi;M. Kondoh;M. Fujii;T. Hayakawa;Yoshiteru Watanabe
基于Ad 5型的重组腺病毒（Ad）载体已广泛用于基因转移实验。传统的Ad 5型载体的向性范围较窄，并且受到可包装的转基因大小的限制。为了克服这些限制，我们之前开发了包含嵌合 Ad 5 型和 35 纤维蛋白的 Ad 载体（Ad5/35 载体）。在当前的研究中，我们评估了Ad5/35载体将基因转移到人滋养层细胞系（JAR、JEG-3和BeWo细胞）中的能力，这些细胞被用作人胎盘的体外模型。我们比较了 Ad5/35 与传统 Ad 载体的基因转移效率。我们发现，通过流式细胞术测定，在所有 3 种滋养层细胞系中，Ad5/35 载体受体 CD46 的表达高于柯萨奇病毒和腺病毒受体的表达。接下来，我们比较了各自表达荧光素酶作为报告基因的Ad5载体和Ad5/35载体的转导活性。 Ad5/35载体在所有3个滋养层细胞系中均具有比常规Ad载体更高的基因转移活性（JAR细胞中1.82倍，BeWo细胞中5.37倍，JEG-3细胞中6.11倍）。因此，含有嵌合型 5 和 35 纤维蛋白的 Ad 载体可以成为人类滋养层细胞系中基因转移实验的有力工具。
Recombinant adenovirus (Ad) vectors based on Ad type 5 have been widely used for gene transfer experiments. Conventional Ad type 5 vectors have a narrow range of tropism and are limited by the size of the transgene that can be packaged. To overcome these limitations, we previously developed an Ad vector (Ad5/35 vector) containing a chimeric Ad type 5 and 35 fiber protein. In the current study, we evaluated the ability of the Ad5/35 vector to transfer genes into human trophoblast cell lines (JAR, JEG-3 and BeWo cells), which are used as in vitro models of human placenta. We compared the gene transfer efficiency of Ad5/35 to that of conventional Ad vector. We found that expression of CD46, which are receptors for Ad5/35 vector, are higher than that of coxsackievirus and adenovirus receptor in all 3 trophoblast cell lines, as determined by flow cytometry. Next, we compared the transducing activity of Ad5 vector and Ad5/35 vector that each expressed luciferase as a reporter gene. Ad5/35 vector had greater gene transfer activity than the conventional Ad vector in all 3 trophoblast cell lines (1.82-fold in JAR cells, 5.37-fold in BeWo cells, 6.11-fold in JEG-3 cells). Thus, Ad vector that contains chimeric type 5 and 35 fiber protein can be a powerful tool for gene transfer experiments in human trophoblast cell lines.