J Vet Sci.  2014 Mar;15(1):99-109. 10.4142/jvs.2014.15.1.99.

Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine

• Tang D
• Liu J
• Li C
• Zhang H
• Ma P
• Luo X
• Zeng Z
• Hong N
• Liu X
• Wang B
• Wang F
• Gan Z
• Hao F

• KMID: 1737615
• DOI: http://doi.org/10.4142/jvs.2014.15.1.99

Abstract

The purpose of this study was to investigate the effects of porcine interleukin (IL)-2 and IL-4 genes on enhancing the immunogenicity of a porcine reproductive and respiratory syndrome virus ORF5 DNA vaccine in piglets. Eukaryotic expression plasmids pcDNA-ORF5, pcDNA-IL-2, and pcDNA-IL-4 were constructed and then expressed in Marc-145 cells. The effects of these genes were detected using an indirect immunofluorescent assay and reverse transcription polymerase chain reaction (RT-PCR). Characteristic fluorescence was observed at different times after pcDNA-ORF5 was expressed in the Marc-145 cells, and PCR products corresponding to ORF5, IL-2, and IL-4 genes were detected at 48 h. Based on these data, healthy piglets were injected intramuscularly with different combinations of the purified plasmids: pcDNA-ORF5 alone, pcDNA-ORF5 + pcDNA-IL-2, pcDNA-ORF5 + pcDNA-IL-4, and pcDNA-ORF5 + pcDNAIL-4 + pcDNA-IL-2. The ensuing humoral immune responses, percentages of CD4+ and CD8+ T lymphocytes, proliferation indices, and interferon-gamma expression were analyzed. Results revealed that the piglets co-immunized with pcDNA-ORF5 + pcDNA-IL-4 + pcDNA-IL-2 plasmids developed significantly higher antibody titers and neutralizing antibody levels, had significantly increased levels of specific T lymphocyte proliferation, elevated percentages of CD4+ and CD8+ T lymphocytes, and significantly higher IFN-gamma production than the other inoculated pigs (p < 0.05).

Keyword

IL-2; IL-4; immunogenicity; ORF5 DNA vaccine; porcine reproductive and respiratory syndrome virus

MeSH Terms

Animals
Cell Line
Escherichia coli/genetics
Haplorhini
Immunity, Cellular
Interleukin-2/genetics/*metabolism
Interleukin-4/genetics/*metabolism
Neutralization Tests/veterinary
Plasmids
Porcine Reproductive and Respiratory Syndrome/*prevention & control
Porcine respiratory and reproductive syndrome virus/*immunology
Recombinant Proteins/genetics/metabolism
Swine
Vaccines, DNA/immunology
Viral Envelope Proteins/*genetics/metabolism
Viral Vaccines/*immunology
Interleukin-2
Interleukin-4
Recombinant Proteins
Vaccines, DNA
Viral Envelope Proteins
Viral Vaccines

Figure

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  Fig. 1 Expression of pcDNA-ORF5 after transfection. Scale bars = 100 µm.

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  Fig. 2 RT-PCR results for the transfected cells. M, DL2000 DNA marker; Lanes 1, 3, and 5, RT-PCR products from cells transfected with pcDNA3.1(+); Lane 2, RT-PCR product from cells transfected with pcDNA-IL-2; Lane 4, RT-PCR product from cells transfected with pcDNA-IL-4; Lane 6, RT-PCR product from cells transfected with pcDNA-ORF5.

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  Fig. 3 Detection of recombinant proteins encoded by expression plasmids pcDNA-ORF5(A), pcDNA-IL-2 (B), and pcDNA-IL-4 (C) using Western blotting.

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  Fig. 4 Humoral immune responses in vaccinated piglets from the different groups detected with a commercial ELISA kit.

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  Fig. 5 T lymphocyte proliferation assay results for the immunized piglets.

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  Fig. 6 Serum IFN-γ production in the vaccinated groups and control group.

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