J Bacteriol Virol.  2012 Sep;42(3):224-231. 10.4167/jbv.2012.42.3.224.

Adenovirus Expressing Human Interferon Inhibits Replication of Foot and Mouth Disease Virus and Reduces Fatal Rate in Mice

• Chu JQ
• Kim SM
• Lee KN
• Oem JK
• Ko YJ
• Lee HS
• Kim YJ
• Park JY
• Kim KJ
• Parida S
• Oh Y
• Paton DJ
• Joo YS
• Kim B
• Park JH

• KMID: 1434751
• DOI: http://doi.org/10.4167/jbv.2012.42.3.224

Abstract

Interferon is an important cytokine that plays a critical role in the initial host defense against viral infection. Recombinant human adenoviruses expressing human interferon-alpha (Ad-HIFNalpha) or pig interferon-beta fused with interleukin-18 (Ad-PIFNbeta-IL18) were constructed and used to induce an early protective response against foot and mouth disease (FMD). To analyze the antiviral effect, bovine thyroid and porcine kidney IBRS-2 cells and ICR mice were treated with Ad-HIFNalpha, Ad-PIFNbeta-IL18, and cocktail of Ad-HIFNalpha and Ad-PIFNbeta-IL18. The survival rate of suckling mice was monitored after foot and mouth disease virus (FMDV) challenge following intra-peritoneal (IP) administration of appropriate adenovirus. Indirect antigen ELISA was performed to evaluate inhibition of FMDV replication following challenge with the FMDV O, A, or Asia 1 serotypes in vitro. These recombinant adenoviruses reduced the replication of FMDV in susceptible cells, thereby decreasing the fatality in mice, suggesting that they can be a useful control method for the early protection against FMD infection in livestock after field trial.

Keyword

FMD; Interferon; Protection; Adenovirus

MeSH Terms

Adenoviridae
Adenoviruses, Human
Animals
Asia
Enzyme-Linked Immunosorbent Assay
Foot
Foot-and-Mouth Disease
Foot-and-Mouth Disease Virus
Humans
Interferon-alpha
Interferon-beta
Interferons
Interleukin-18
Kidney
Livestock
Mice
Mice, Inbred ICR
Survival Rate
Thyroid Gland
Interferon-alpha
Interferon-beta
Interferons
Interleukin-18

Figure

• 

  Figure 1 Construction of recombinant adenovirus expressing HIFNα or PIFNβ-IL18. (A) Schematic diagram of genome organization of recombinant adenovirus, Ad5; adenovirus serotype 5, LITR; left inverted terminal repeat, RITR; right inverted terminal repeat, ΔE1; deletion of E1 gene, ΔE3; deletion of E3 gene, pA; β-globin poly A, MLP; adenoviral major late promoter, CMV; cytomegalovirus promoter, GFP; green fluorescent protein. (B) Sequences of the adenoviral DNA (Ad-HIFN and Ad-PIFNβ-IL18) in cloning region for foreign gene.

• 

  Figure 2 Inhibition of FMDV replication by anti-viral plasmids in IBRS-2 cells. This result shows inhibition of FMDV replication by transfection of antiviral plasmids in IBRS-2 cells. The plasmids containing HIFNα or PIFNβ/IL18 fusion genes (pHIFNα and pPIFNβ-PIL18) demonstrated anti-FMDV potential. Optical density in only FMDV-infected or mock-infected cells was 3.37±0.0332 or 0.06±0.0035, respectively.

• 

  Figure 3 Western blot analysis of HIFNα, PIFNβ-IL18 expressed by recombinant adenovirus in HEK 293A cells. Lysates of HEK 293A cells infected with recombinant adenovirus were analyzed by transferring to nitrocellulose membranes for Western blot analysis. Lane M; protein molecular weight marker (BIO-RAD). Lane 1; cell lysates from Ad-HIFNα infected HEK 293A cells. Lane 2; supernatant from Ad-HIFNα infected HEK 293A cell. Lane 3, 7; cell lysate from mock-infected HEK 293A cell. Lane 4, 8; supernatant of mock-infected HEK 293A cell. Lane 5; cell lysate of Ad-PIFNβ-PIL18 infected HEK 293A cell. Lane 6; supernatant from Ad-PIFNβ-PIL18 infected HEK 293A cell.

• 

  Figure 4 Anti-FMDV activity by recombinant adenoviruses in bovine and porcine-originated cells. FMDV antigen ELISA was determined from the supernatant taken at 72 h after FMDV infection. Cells were infected with Ad-HIFNα or Ad-PIFNβ-IL18 (104.0 TCID50/0.1 ml) and FMDV serotypes (102.0 TCID50/0.1 ml) of O/UKG, Asia1/ISR and A22/Iraq was used to challenge cells at 24 h after infection with the recombinant adenoviruses. The control group for adenovirus control (Ad-control) was inoculated adenovirus expressing lamin-shRNA.

• 

  Figure 5 Fatality in suckling mice following adenovirus treatment and FMDV challenge. (A) Mice were challenged with FMDV (20 LD50/0.1 ml) at 24 h after recombinant adenovirus (106 TCID50/0.1 ml) inoculation by intraperitoneal (IP) route. (B) Mice were challenged with FMDV O/SKR/2002 (125 LD50/0.1 ml) at 24 h. To determine synergic effect by co-administration with adenoviruses, high dose of FMDV (125 LD50/0.1 ml) and adenovirus (Ad-HIFNα, 106.5 TCID50/0.1 ml and Ad-PIFNβ-IL18, 107.5 TCID50/0.1 ml) were administrated to suckling ICR mice by intraperitoneal (IP) route. The control group (control) was only inoculated with FMDV.

• 

  Figure 6 Change of fatality in suckling mice following several adenovirus administration and FMDV challenge. Recombinant adenoviruses (106.0 TCID50/0.1 ml) were inoculated by intraperitoneal (IP) route at -1, 3 DPC (2 round injection group, 2X) and at -1, 0, 3 and 6 DPC (4 round injection group, 4X) in sucking ICR mice. The mice were 7-day old when challenged by FMDV O/SKR/2002 (20 LD50/mouse). The control group for adenovirus (Ad-con) was inoculated adenovirus expressing lamin-shRNA. The control group (control) was only inoculated with FMDV.

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