一种高效感染血液细胞的新型靶向性腺病毒载体的构建.pdfVIP

21 2 21 2106-110 2006 3 VIROLOGICA SINICA March 2006 ∗ 1,2 2 2∗∗ 1 1 ∗∗ (1. 230032 2. 100850) Construction of a Novel Retargeted Adenoviral Vector with High Gene Transfer Efficiency into Hematopoietic Cells 1,2 2 2∗∗ 1 1∗∗ NI Fang , LU Zhuo-zhuang , WANG Li-sheng , QU Cheng-kui , WANG Si-ying (1. Department of Pathophysiology, Anhui Medical University, Hefei 230032, China; 2. Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China) Abstract: The adenovirus subgenus C serotypes 5 (Ad5) vectors which have been used to transduce epithelial cells have very limited ability to infect hematopoietic cells. 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 Ad11p fiber shaft and knob domains. An Ad5-based vector encoding the green fluorescent protein (GFP) gene under the control of the CMV promoter with Ad11p fiber receptor specificity was generated (Ad5F11p-GFP). The Ad5F11p-GFP vector-mediated gene transfer efficiency for some committed hematopoietic cell lines such as myeloblasts (K562) and monocytes (U937) was evaluated using flow cytometry and compared to that of Ad5-GFP, which also encodes the GFP gene under the control of the CMV promoter. The results showed that these cell lines were superiorly transduced by the Ad5F11p-GFP vector at the same MOI (multiplicity of infection, MOI) compared with the Ad5-GFP vectormore than 90% tested cells were infected by Ad5F11p-GFP adenovirus