J Vet Sci.  2014 Sep;15(3):399-407. 10.4142/jvs.2014.15.3.399.

Enhancing mucosal immunity in mice by recombinant adenovirus expressing major epitopes of porcine circovirus-2 capsid protein delivered with cytosine-phosphate-guanosine oligodeoxynucleotides

• Chang HT
• He XY
• Liu YF
• Chen L
• Guo QH
• Yu QY
• Zhao J
• Wang XW
• Yang X
• Wang CQ

• KMID: 2155623
• DOI: http://doi.org/10.4142/jvs.2014.15.3.399

Abstract

A recombinant replication-defective adenovirus expressing the major epitopes of porcine circovirus-2 (PCV-2) capsid protein (rAd/Cap/518) was previously constructed and shown to induce mucosal immunity in mice following intranasal delivery. In the present study, immune responses induced by intranasal immunization with a combination of rAd/Cap/518 and cytosine-phosphate-guanosine oligodeoxynucleotides (CpG ODN) were evaluated in mice. The levels of PCV-2-specific IgG in serum and IgA in saliva, lung, and intestinal fluids were significantly higher in the group immunized with rAd/Cap/518 and CpG ODN than animals immunized with rAd/Cap/518 alone. The frequencies of IL-2-secreting CD4+ T cells and IFN-gamma-producing CD8+ T cells were significantly higher in the combined immunization group than mice immunized with rAd/Cap/518 alone. The frequencies of CD3+, CD3+CD4+CD8-, and CD3+CD4-CD8+ T cells in the combined immunization group were similar to that treated with CpG ODN alone, but significantly higher than mice that did not receive CpG ODN. PCV-2 load after challenge in the combined immunization group was significantly lower than that in the phosphate-buffered saline placebo group and approximately 7-fold lower in the group treated with CpG ODN alone. These results indicate that rAd/Cap/518 combined with CpG ODN can enhance systemic and local mucosal immunity in mice, and represent a promising synergetic mucosal vaccine against PCV-2.

Keyword

cytosine-phosphate-guanosine oligodeoxynucleotides; mucosal immunity; porcine circovirus-2; rAd/Cap/518

MeSH Terms

Adenoviridae/genetics/immunology
Administration, Intranasal
Animals
Capsid Proteins/*genetics/immunology
Circoviridae Infections/*immunology
Circovirus/*genetics/immunology
Epitopes/genetics/immunology
Female
Immunity, Mucosal/immunology
Immunoglobulin A/blood/immunology
Immunoglobulin G/blood/immunology
Mice
Mice, Inbred BALB C
Oligodeoxyribonucleotides/genetics
Vaccines, Synthetic/genetics/immunology
Viral Vaccines/administration & dosage/*genetics/immunology
Capsid Proteins
Epitopes
Immunoglobulin A
Immunoglobulin G
Oligodeoxyribonucleotides
Vaccines, Synthetic
Viral Vaccines

Figure

• 

  Fig. 1 Levels of PCV-2-specific IgG in serum (A) or IgA in saliva (B) along with lung (C) and intestinal (D) lavage fluid from mice (n = 5) immunized with rAd/Cap/518 plus CpG ODN through an intranasal route on the indicated sampling day. Identical letters on top of the bars at each time point indicate no significant difference (p > 0.05). Different letters on top of the bars indicate a significant difference (p < 0.05).

• 

  Fig. 2 The percentages of T cell subsets among spleen lymphocytes in mice 14 days after boosting immunization. Identical letters on top of the bars indicate no significant difference (p > 0.05). Different letters on top of the bars indicate significant differences (p < 0.05).

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  Fig. 3 Representative FACS results showing the percentages of IL-2-secreting CD4+ T cells and IFN-γ-secreting CD8+ T cells among spleen lymphocytes measured 14 days after boosting immunization.

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  Fig. 4 qPCR result showing the PCV-2 viral genomic copy loads in spleen, lung, and lymph node samples collected from mice 14 days after viral challenge. Identical letters on top of the bars indicate no significant difference (p > 0.05). Different letters on top of the bars indicate significant differences (p < 0.05).

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