Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (10): 2058-2066.doi: 10.3864/j.issn.0578-1752.2012.10.019

• VETERINARY SCIENCE • Previous Articles     Next Articles

Immune Enhancing Effects of Canine Interleukin-7 Gene on Canine Parvovirus DNA Vaccine

 SUN  Yan, ZHONG  Fei, LI  Xiu-Jin, WANG  Xing-Xing, WANG  Lu, JIA  Qi-Heng, HAN  Dong-Mei, LI  Zhen, ZHANG  Feng, PAN  Hong-Li   

  1. 1.河北农业大学动物医学院基础兽医部/农业部动物疫病病原生物学华北区观测实验站,河北保定071001
    2.燕山大学环境与化学工程学院生物工程系,河北秦皇岛 066004
  • Received:2011-12-23 Online:2012-05-15 Published:2012-04-06

Abstract: 【Objective】 Animal interleukin -7 (IL-7), a multifunctional cytokine, plays an important role in B- and T-cell generation and development and in improvement of animal immune capacity coordinating with other cytokines. In this study, the immune enhancing effects of canine IL-7 (cIL-7) gene was investigated in mice co-immunized with expression vectors of canine parvovirus VP2 DNA vaccines. 【Method】Canine IL-7 genes, both with stop codon and without stop codon, were amplified from canine splenic lymphocytes by RT-PCR, and then inserted into the eukaryotic expression vector pcDNA3.1A to contruct the non-fused and Myc/His-tag-fused cIL-7 eukaryotic secretory expression vectors (pcDNA-cIL7 and pcDNA-cIL7/MH), respectively. To determine whether the expression vector could mediate cIL-7 gene expression in eukaryotic cells, the pcDNA-cIL7/MH plasmid was transfected into HEK293T cells using calcium phosphate transfection method for transient expression. The mice were co-immunized with VP2 gene expression vector (pcDNA-CD5-VP2, constructed previously in authors’ laboratory) and cIL-7 gene expression vector pcDNA-cIL7, the mice immunized with either pcDNA-cIL7 or pcDNA-CD5-VP2 were used as the control. After immunization, the antibodies against CPV in the immunized mice at different times were measured by ELISA. The spleen lymphocyte proliferation response at 35 d post-immunization was determined by lymphocyte proliferation assay, and the interferon-γ expression level of the mouse lymphocytes was measured by ELISA. 【Results】 The results showed that the sequence of cIL-7 gene amplified was consistent with that in GenBank. Western-blot showed that the recombinant cIL-7 could be expressed and secreted in HEK293T cells. Immunization results showed that the antibody levels and the neutralizing antibody titers in the serum of VP2/cIL-7-immunized mice were significantly higher than that of VP2-immunized mice (P<0.01 and P<0.05), respectively. The lymphocyte stimulation indexes and secreted IFN-γ levels of the VP2/cIL-7-immunized mice were significantly higher than that of VP2-imunized mice (P<0.05), respectively. 【Conclusion】The cIL-7 gene can significantly enhance the immune response of the mice to CPV VP2 DNA vaccine.

Key words: canine interleukin -7, canine parvovirus, VP2 gene, DNA vaccine

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