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中国生物工程杂志

China Biotechnology
China Biotechnology  2011, Vol. 31 Issue (06): 86-92    DOI:
    
The Characterization of Modified Adenovirus Vector
QIAO Wei1,2, HE Jin-shen1,2, FU Yuan-hui2, HONG Tao2,3
1. Department of Immunology, Anhui Medical University, Hefei 230032, China;
2. College of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing 100044, China;
3. Institute of Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China
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Abstract  

Objective: A simple way to construct an adenovirus vector containing a peptide-RGD in the HI loop of the fiber knob was developed and then its infection efficiency on RD and T24 cells in vitro and the humoral responses to transgenes in vivo were investigated. Methods: The pAdEasy-RGD-1 was modified by inserting RGD motif into pAdEasy-1 based on simple PCR method. Enhanced green fluorescent protein (EGFP) and codon optimized fusion protein (Fsyn) of human respiratory syncytial virus (RSV) were cloned and constucted into pAdEasy-RGD-1 to generate adenovirus vectors of FGAd-RGD-EGFP and FGAd-RGD-Fsyn, respectively. FGAd-RGD-EGFP infected RD and T24 cells to investigate its infection efficiency using fluorescent microscope and flow cytometry. FGAd-RGD-Fsyn was exploited to vaccinate BALB/c mice via intramuscular and imtranasal routes, respectively, to evaluate humoral responses to transgenes in vivo. Results: The infection efficiency on RD and T24 cells was higher with FGAd-RGD-EGFP than FGAd-EGFP, and immunization with FGAd-RGD-Fsyn generated similar serum IgG specific for RSV-F. Conclusions: PCR is a simple and practical method to construct adenovirus vector containing a peptide-RGD in the HI loop of the fiber knob. The resultant modified adenovirus vector displays improved infection efficiency on RD and T24 cells and similar humoral immune responses to transgene as compared to original adenovirus vector. Therefore, it will be an useful platform for oncotherapy.



Key wordsReplication-defective adenovirus vector      RGD      Fluorensence      Humoral immune response     
Received: 15 November 2010      Published: 28 June 2011
ZTFLH:  Q819  
Cite this article:

QIAO Wei, HE Jin-shen, FU Yuan-hui, HONG Tao. The Characterization of Modified Adenovirus Vector. China Biotechnology, 2011, 31(06): 86-92.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2011/V31/I06/86

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