摘要
本试验目的是研究牛IL-2重组质粒对口蹄疫病毒(Foot-and-mouth disease virus,FMDV)G-H环多肽免疫小鼠的佐剂效应。作者构建了含有信号肽序列的牛IL-2真核表达质粒pcDNA3.1-pBoIL-2,并体外鉴定其表达,同时原核表达并纯化了Asia1型FMDV G-H环多肽。采用不同组合的pcDNA3.1-pBoIL-2、FMDV G-H环多肽分组免疫小鼠,并设阴、阳性对照,通过间接ELISA试验、微量血清中和试验、淋巴细胞增殖试验和实时定量PCR试验对各免疫组小鼠体液和细胞免疫水平进行综合评价。结果表明,与G-H+pcDNA3.1免疫组比较,G-H+pcD-NA3.1-pBoIL-2免疫组特异性抗体水平、中和抗体滴度、淋巴细胞增殖水平和细胞因子相对表达水平显著提高(P<0.05)。结果提示,牛IL-2真核表达质粒可以显著提高G-H环多肽的免疫效果。
To study the immune response induced by G-H loop peptide of Asia1 foot-and-mouth disease virus（FMDV） with BoIL-2 recombinant plasmid as adjuvant,the immune effects in immunized mice were investigated.Recombinant plasmids pcDNA3.1-pBoIL-2 was constructed and the condition of the expression was identified in vitro.G-H peptide was expressed in Rosetta（DE3）pLysS and were successfully purified through GST affinity purification.Five groups of Kunming mice each with 8 were inoculated with different combinations including PBS,pcDNA3.1-pBoIL-2 and/or G-H peptide,subsequently,immune effects have been evaluated through indirect ELISA assay,the neutralization test,lymphocytes proliferation assay and Quantitive Real-time PCR.The results showed that in comparing with the group immunized with G-H＋pcDNA3.1,the group communication with pcDNA3.1-pBoIL-2 induced significantly humoral and cellular immune responses.The results of this study indicate that the co-inoculation of G-H loop peptide with rBoIL-2（as adjuvant） showed good immune effect.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2009年第11期1651-1657,共7页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家科技支撑计划课题(2006BAD06A17-08)
黑龙江省十一五重大科技项目(GA06B202)
国家奶牛技术体系
