中国农业科学2012，Vol.45Issue(10)：2058-2066,9.DOI:10.3864/j.issn.0578-1752.2012.10.019
犬白细胞介素-7基因对犬细小病毒DNA疫苗的免疫增强作用
Immune Enhancing Effects of Canine Interleukin-7 Gene on Canine Parvovirus DNA Vaccine
摘要
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 contract 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 EL1SA. 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.关键词
犬IL-7/细小病毒/VP2基因/DNA疫苗Key words
canine interleukin -7/ canine parvovirus/ VP2 gene/ DNA vaccine引用本文复制引用
孙岩,潘红丽,仲飞,李秀锦,王幸兴,王璐,贾启恒,韩冬梅,李振,张峰..犬白细胞介素-7基因对犬细小病毒DNA疫苗的免疫增强作用[J].中国农业科学,2012,45(10):2058-2066,9.基金项目
国家自然科学基金(30771586) （30771586）
河北省自然科学基金(C2008000244) （C2008000244）
