J Vet Sci.  2006 Sep;7(3):257-262. 10.4142/jvs.2006.7.3.257.

Enhanced immune response with foot and mouth disease virus VP1 and interleukin-1 fusion genes

• Park JH
• Kim SJ
• Oem JK
• Lee KN
• Kim YJ
• Kye SJ
• Park JY
• Joo YS

• KMID: 1089910
• DOI: http://doi.org/10.4142/jvs.2006.7.3.257

Abstract

The capsid of the foot and mouth disease (FMD) virus carries the epitopes that are critical for inducing the immune response. In an attempt to enhance the specific immune response, plasmid DNA was constructed to express VP1/interleukin-1alpha (IL-1alpha) and precursor capsid (P1) in combination with 2A (P1-2A)/IL-1alpha under the control of the human cytomegalovirus (HCMV) immediateearly promoter and intron. After DNA transfection into MA104 (monkey kidney) cells, Western blotting and an immunofluorescence assay were used to confirm the expression of VP1 or P1-2A and IL-1alpha. Mice were inoculated with the encoding plasmids via the intradermal route, and the IgG1 and IgG2a levels were used to determine the immune responses. These results show that although the immunized groups did not carry a high level of neutralizing antibodies, the plasmids encoding the VP1/ IL-1alpha, and P1-2A /IL-1alpha fused genes were effective in inducing an enhanced immune response.

Keyword

foot and mouth disease; immunogenicity; interleukin-1

MeSH Terms

Animals
Antibodies, Viral/blood
Capsid Proteins/biosynthesis/genetics/*immunology
Cell Line
DNA, Viral/genetics
Enzyme-Linked Immunosorbent Assay
Foot-and-Mouth Disease/*immunology/prevention&control
Foot-and-Mouth Disease Virus/genetics/*immunology
Haplorhini
Immunization
Interleukin-1/biosynthesis/genetics/*immunology
Male
Mice
Mice, Inbred C57BL
Plasmids/genetics
Polymerase Chain Reaction
Recombinant Fusion Proteins/biosynthesis/genetics/immunology
Specific Pathogen-Free Organisms
Transfection
Vaccines, DNA/genetics/*immunology

Figure

• 

  Fig. 1 Schematic diagram of plasmid constructs expressing various FMDV proteins in the DNA-based mammalian expression vectors pCMV: human cytomegalovirus immediate-early promoter. SV40 p(A): SV40 polyadenylation signal.

• 

  Fig. 2 Detection of recombinant proteins in pS-VP1, pSIL1A-VP1 and pSIL1A-P12A transfected cells by immunofluorescent assay with bovine anti-FMDV serum. A. pSLIA transfected MA104 cells as a control, B. pS-VP1 transfected MA104 cells, C. pSIL1A-VP1 transfected MA104 cells, D. pSIL1A-P12A transfected MA104 cells.

• 

  Fig. 3 Expression of recombinant proteins in transfected cells as detected by Western blot analysis with swine IL-1α antibody (A) or bovine FMDV-antiserum (B). Lysates of MA104 cells transfected with recombinant plasmids were analysis by electrophoretically transferred to nitrocellulose membrane for Western blot analysis. M; Protein molecular weight marker (Invitrogen), lane 1; pSLIA transfected MA104 cells as a control, lane 2; pS-VP1 transfected MA104 cells, lane 3; pSIL1A-VP1 transfected MA104 cells, lane 4; pSIL1A-P12A transfected MA104 cells.

• 

  Fig. 4 Immune responses in the mice immunized with FMDV plasmids. (A) The ELISA was carried out with the sera from C57BL/6 mice (five per group). The test was determined at 1 week after the third immunization. The mice immunized with recombinant plasmids encoding IL-1α showed high titers. The optical densitiy was measured at 405 nm. (B) C57BL/6 mice (five per group) were immunized three times with 10 µl of plasmids by tail injection. ELISA was carried out with each serum from five mice. The test sera from the vaccinated animals were diluted 1 : 300 and optical density read at 405 nm. Similarly increased immune response in the pSIL1A-VP1 and pSIL1A-P12A group were detected in the sera at one week after the 2nd immunization.

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  Fig. 5 Levels of IgG subclass in the mice immunized with FMDV plasmids. The T cell response was tested by IgG subclass analysis in the sera from C57BL/6 mice (five per group) at one week after 3rd immunization. The test sera from the vaccinated animals were diluted 1 : 50 and optical density read at 405 nm. The mice immunized with pSIL1A-VP1 and pSIL1A-P12A elicited specific IgG1 and IgG2a response and increased the ratio of IgG1/IgG2a.

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