Ghaemi Amir, Soleimanjahi Hoorieh, Gill Pooria, Hassan Zuhair, Jahromi Soodeh Razeghi M, Roohvand Farzin
Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, 14115-111 Iran.
Genet Vaccines Ther. 2010 May 12;8:3. doi: 10.1186/1479-0556-8-3.
Lambda phages have considerable potential as gene delivery vehicles due to their genetic tractability, low cost, safety and physical characteristics in comparison to other nanocarriers and gene porters. Little is known concerning lambda phage-mediated gene transfer and expression in mammalian hosts. We therefore performed experiments to evaluate lambda-ZAP bacteriophage-mediated gene transfer and expression in vitro. For this purpose, we constructed recombinant lambda-phage nanobioparticles containing a mammalian expression cassette encoding enhanced green fluorescent protein (EGFP) and E7 gene of human papillomavirus type 16 (lambda-HPV-16 E7) using Lambda ZAP- CMV XR vector. Four cell lines (COS-7, CHO, TC-1 and HEK-239) were transduced with the nanobioparticles. We also characterized the therapeutic anti-tumor effects of the recombinant lambda-HPV-16 E7 phage in C57BL/6 tumor mice model as a cancer vaccine. Obtained results showed that delivery and expression of these genes in fibroblastic cells (COS-7 and CHO) are more efficient than epithelial cells (TC-1 and HEK-239) using these nanobioparticles. Despite the same phage M.O.I entry, the internalizing titers of COS-7 and CHO cells were more than TC-1 and HEK-293 cells, respectively. Mice vaccinated with lambda-HPV-16 E7 are able to generate potent therapeutic antitumor effects against challenge with E7- expressing tumor cell line, TC-1 compared to group treated with the wild phage. The results demonstrated that the recombinant lambda-phages, due to their capabilities in transducing mammalian cells, can also be considered in design and construction of novel and safe phage-based nanomedicines.
与其他纳米载体和基因转运体相比，λ噬菌体因其遗传易操作性、低成本、安全性和物理特性，作为基因传递载体具有相当大的潜力。关于λ噬菌体介导的基因在哺乳动物宿主中的转移和表达，人们了解甚少。因此，我们进行了实验以评估λ-ZAP噬菌体在体外介导的基因转移和表达。为此，我们使用Lambda ZAP-CMV XR载体构建了重组λ噬菌体纳米生物颗粒，其包含编码增强型绿色荧光蛋白（EGFP）和人乳头瘤病毒16型E7基因（λ-HPV-16 E7）的哺乳动物表达盒。用这些纳米生物颗粒转导了四种细胞系（COS-7、CHO、TC-1和HEK-239）。我们还将重组λ-HPV-16 E7噬菌体作为癌症疫苗，在C57BL/6肿瘤小鼠模型中表征了其治疗性抗肿瘤作用。获得的结果表明，使用这些纳米生物颗粒，这些基因在成纤维细胞（COS-7和CHO）中的递送和表达比上皮细胞（TC-1和HEK-239）更有效。尽管噬菌体的感染复数相同，但COS-7和CHO细胞的内化滴度分别高于TC-1和HEK-293细胞。与用野生噬菌体处理的组相比，用λ-HPV-16 E7疫苗接种的小鼠能够对用表达E7的肿瘤细胞系TC-1进行的攻击产生有效的治疗性抗肿瘤作用。结果表明，重组λ噬菌体由于其转导哺乳动物细胞的能力，也可用于设计和构建新型安全的基于噬菌体的纳米药物。
