摘要
将猪白细胞介素2(pIL-2)与猪白细胞介素6(pIL-6)的cDNA序列克隆到大肠杆菌表达载体pET-28a及pGEX-KG上,转化BL21(DE3)表达菌,经IPTG诱导产生重组蛋白pIL-2与pIL-6,表达量分别占菌株总蛋白的43.45％和28.37％,经提纯后含量可分别达到97.06％和86.37％。将提纯后不同剂量的两种重组蛋白以单独或组合的方式,加入伪狂犬病(Ea株)TK^-/gG^-双基因缺失疫苗中,使每头份疫苗分别含2、5和10μg·ml^(-1)的pIL-2或(和)pIL-6,用此疫苗免疫伪狂犬病抗体阴性猪。同时设试验对照组。初次免疫30d后加强免疫,测定中和抗体,观察试验猪的日增重情况,进行统计分析。发现各试验组的抗体水平均高于不含重组蛋白的基因缺失疫苗组;其中,含2μg·ml^(-1)pIL-2试验组与含10μg·ml^(-1)pIL-2/pIL-6试验组抗体水平和基因缺失疫苗对照组之间呈显著差异(P=0.019)与极显著差异(P=0.009)。结果表明,大肠杆菌表达的pIL-2、pIL-6对伪狂犬病鄂A株基因缺失疫苗(TK^-/gG^-/LacZ^+)有较强的佐剂效应,且呈剂量相关性。不同试验组日增重比较结果表明,免疫效果高低与猪的生长速度呈正相关。本试验为重组pIL-2与pIL-6作为新型疫苗佐剂应用提供了重要的试验依据。
Porcine interleukin-2 and porcine interleukin-6 cDNA sequences were cloned into the expressing plasmid vectors pET-28a and pGEX-KG, respectively; and expressed in E. coli BL21(DE3) with high-level production. The gene deleted vaccine of pseudorabies virus Ea strain ( TK- /gG- /LacZ + ) as mixed with the two different purified recom-binant proteins each, or both, with the doses of 2, 5 and 10ug.ml-1. Ten groups of pseudorabies negative antibody swines were immunized twice with tested vaccines at different doses, or control vaccine, respectively. The antibody titers of the test groups were delected by neulralizalion test, and the daily weight gains of swines were calculated and analyzed statistically. It was found that all the neutralizing anlibody titers in lest groups were higher than the control group, and the recombinant proleins appeared a dose dependenl adjuvant effect. A significant and extremely significant differences, were observed in tested vaccines containing 2 ug.ml-1 pIL-2 and with 10ug.ml-1 pIL-2/pIL-6, and the vaccines with-out pILs. The difference of the daily weighl gain indicated the potenlial posilive influence of pIL-2 and pIL-6 on immune proteclion.
出处
《中国农业科学》
CAS
CSCD
北大核心
2003年第10期1213-1218,共6页
Scientia Agricultura Sinica
基金
国家"863"计划资助项目(2001AA213051)
