J Vet Sci.  2015 Sep;16(3):265-272. 10.4142/jvs.2015.16.3.265.

Establishment and evaluation of a murine alphavbeta3-integrin-expressing cell line with increased susceptibility to Foot-and-mouth disease virus

Affiliations
  • 1State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory, Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, La

Abstract

Integrin alphavbeta3 plays a major role in various signaling pathways, cell apoptosis, and tumor angiogenesis. To examine the functions and roles of alphavbeta3 integrin, a stable CHO-677 cell line expressing the murine alphavbeta3 heterodimer (designated as "CHO-677-malphavbeta3" cells) was established using a highly efficient lentiviral-mediated gene transfer technique. Integrin subunits alphav and beta3 were detected at the gene and protein levels by polymerase chain reaction (PCR) and indirect immunofluorescent assay (IFA), respectively, in the CHO-677-malphavbeta3 cell line at the 20th passage, implying that these genes were successfully introduced into the CHO-677 cells and expressed stably. A plaque-forming assay, 50% tissue culture infective dose (TCID50), real-time quantitative reverse transcription-PCR, and IFA were used to detect the replication levels of Foot-and-mouth disease virus (FMDV) in the CHO-677-malphavbeta3 cell line. After infection with FMDV/O/ZK/93, the cell line showed a significant increase in viral RNA and protein compared with CHO-677 cells. These findings suggest that we successfully established a stable alphavbeta3-receptor-expressing cell line with increased susceptibility to FMDV. This cell line will be very useful for further investigation of alphavbeta3 integrin, and as a cell model for FMDV research.

Keyword

alphavbeta3; cell line; Foot-and-mouth disease virus; integrin; receptor

MeSH Terms

Animals
Animals, Suckling
CHO Cells
Cloning, Molecular
Cricetulus
DNA, Complementary/genetics/metabolism
Disease Susceptibility/virology
Foot-and-Mouth Disease/*genetics/virology
Foot-and-Mouth Disease Virus/*physiology
Integrin alphaVbeta3/*genetics/metabolism
Mice
DNA, Complementary
Integrin alphaVbeta3
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