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Journal of Animal and Veterinary Advances

Year: 2013  |  Volume: 12  |  Issue: 1  |  Page No.: 108 - 117

Establishment of a Murine αυβ1 Transgenic CHO-K1 Cell Line and its Susceptibility to Foot-and-Mouth Disease Virus Type Asia l/HN/2006 in China

Y. Zhang, H.X. Zheng, Z.D. Zhang, Y. Jin, F. Yang, J.J. He, W.J. Cao, D.H. Sun and L. Lv

Abstract

Field isolates of Foot-and-Mouth Disease Virus (FMDV) were found to use four αυ integrins (αυβ1, αυβ3, αυβ6 and αυβ8) as cellular receptors. Researchers established a stable Chinese Hamster Ovary clone K1 (CHO-K1) cell line expressing the murine αυβ1 heterodimer (designated as CHO-K1-αυβ1) using a highly efficient lentiviral-based gene transfer technology to deliver murine αυ, Internal Ribosome Entry Site (IRES) and β1 genes into cell chromosomes and the inserted genes were then transcribed from a Cytomegalovirus (CMV) promoter. αυβ1 expression was stringently regulated by Doxycycline (Dox) and was found to be stable. CHO-K1-αυβ1 cells were susceptible to FMDV type Asia l/HN/2006. The plaque assay revealed that the virus produced bigger and more plaques in CHO-K1-αυβ1 cells (1.05x104 PFU mL-1) than in CHO-K1 cells. When sodium heparin (1 and 2 mg mL-1) was used as the inhibitor, the number of plaques in CHO-K1 cells were significantly decreased (4.0x103-35 and 20 PFU mL-1), supported by time-course of replication and proliferation. The number and size of plaques on CHO-K1-αυβ1 cells showed no obvious change, indicating that the αυβ1 heterodimer expressed on CHO-K1-αυβ1 can be used as an FMDV receptor.

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