J Bacteriol Virol.  2020 Mar;50(1):35-43. 10.4167/jbv.2020.50.1.035.

Stable Expression of Bovine Integrin Beta-6 Increases Susceptibility of Goat Kidney Cell Line to Foot-and-mouth Disease Virus

• Kim SM
• Kim SK
• Lee KN
• Park JH
• Kim B

• KMID: 2471836
• DOI: http://doi.org/10.4167/jbv.2020.50.1.035

Abstract

The integrins αvβ1, αvβ3, αvβ6, and αvβ8 are known to be the natural receptors of foot-and-mouth disease virus (FMDV). Among them, integrin αvβ6 is considered a major receptor for FMDV. We performed protein expression of full-length bovine integrins αv, β3, and β6 and confirmed the high efficiency of bovine αvβ6 as the FMDV receptor in FMDV non-permissive SW 480 cells. Next, we established the black goat kidney (BGK) cell line, stably expressing bovine integrin β6 (BGK-β6-4). We observed that BGK-β6-4 cells had significantly enhanced sensitivity to FMDV compared with that of BGK cells (P<0.05). In addition, BGK-β6-4 cells had equal or higher sensitivity to several serotypes of FMDV compared with that of other FMDV permissive cell lines, such as BHK-21 and IBRS-2. In conclusion, we established a promising novel goat cell line, BGK-β6-4, which can be used to isolate or culture FMDV. Furthermore, the BGK-β6-4 cell line may serve as a promising tool for studying integrin αvβ6 receptor functions.

Keyword

Foot-and-mouth disease virus; goat cell line; integrin αvβ6

MeSH Terms

Animals
Cell Line*
Foot-and-Mouth Disease Virus*
Foot-and-Mouth Disease*
Goats*
Integrins
Kidney*
Serogroup
Integrins

Figure

• 

  Figure 1. Detection of bovine integrins αvβ3 and αvβ6 heterodimer expression in SW480 cells using indirect immunofluorescence assay (IFA). SW-480 cells were co-transfected with vectors expressing bovine αv and β6 (a) or β3 (b) subunits. Protein expression was detected by IFA after 48 h of transfection using an anti-human β6 Mab (2076Z) (a) or an anti-human integrin αvβ3 Mab (LM609) (b). Untransfected cells treated with the same primary and secondary antibodies were used as negative controls (c and d).

• 

  Figure 2. Comparison of viral infection efficiency in the cells expressing bovine αvβ3 and αvβ6 integrins. SW480 cells were co-transfected with vectors expressing bovine αv and β6 or β3 subunits. Twenty-four hours after transfection, the cells were infected with 104.5 TCID50 of FMDV O/SKR/2002, Asia 1/M0G/05, or A22/IRQ 24/64 viral suspensions. The supernatant was collected at 8 days after the infections, and viral RNA was extracted. FMDV quantification was performed using real-time RT PCR. An unpaired t-test was performed for statistical analysis by GraphPad InStat software (∗P< 0.05, ∗∗P< 0.01, ∗∗∗P < 0.001). Error bars indicate standard deviations from the mean.

• 

  Figure 3. Comparative virus titration in BGK-β6 stable cell lines. Four stable BGK cell lines, expressing bovine integrin β6 (BGK-β6-1, BGK-β6-2, BGK-β6-3, and BGK-β6-4), in addition to BGK and IBRS-2 cell lines were used for titrating FMDV O/SKR/2002 or C3 Resende. Virus titration was measured 72 h post FMDV infection. TCID50 was calculated by Reed and Muench method (14). An unpaired t-test was performed for statistical analysis by GraphPad InStat software (∗P< 0.05, ∗∗P< 0.01). Error bars indicate standard deviations from the mean.

• 

  Figure 4. Comparative titration of various strains of FMDV in BGK-β6-4, BGK, IBRS-2, and BHK-21 cell lines. BGK-ß6-4, BGK, IBRS-2, and BHK-21 cell lines were used for titrating FMDV A22/IRQ 24/64, A/Pocheon/SKR/2010, Asia1/MOG/05, SAT2/ZIM 5/81, and SAT3/ZIM 4/81. Virus titration was measured 72 h post FMDV infection. TCIID50 was calculated by Reed and Muench method (14). An unpaired t-test was performed for statistical analysis by GraphPad InStat software (∗P< 0.05, ∗∗P< 0.01). Error bars indicate standard deviations from the mean.

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