【摘要】 将转化节瘤拟杆菌 （D .nodosus）纤毛蛋白 （Pili）和绵羊白细胞介素 2 （oviIL2 ）融合基因表达质粒的工程菌BL₂ 1,在含梭苄青霉素营养肉汤培养基中表达 ,离心获得菌体沉淀物 ,裂解后配制成Pili_oviIL2融合基因工程疫苗。用加佐剂和不加佐剂的Pili_oviIL2融合基因工程疫苗分别接种 2只健康家兔 ,2 1天后接种第 2次 ;定期采血 ,用对流免疫电泳检测试验兔的体液免疫反应。结果发现 ,加佐剂和不加佐剂的Pili_oviIL2融合基因工程疫苗免疫兔 7天即可产生相应抗体 ,抗体在免疫血清中可维持 6个月以上。进一步试验将不加佐剂的Pili_oviIL2融合基因工程疫苗接种 3只健康绵羊 ,同样 2 1天后接种第 2次 ,定期采血 ,用对流免疫电泳检测试验绵羊的体液免疫反应。同时用Pili基因工程疫苗接种 2只绵羊作对照。结果 3只绵羊接种Pili_oviIL2融合基因工程疫苗后 ,分别于 7天和 14天产生相应的抗体 ,而接种Pili基因工程疫苗的绵羊于 2 8天产生相应的抗体 ;被免疫绵羊血清中的抗体可维持 6个月以上。用Pili_oviIL2融合基因工程疫苗接种兔和绵羊的免疫试验表明 ,Pili_oviIL2融合基因工程疫苗具有较好的体液免疫应答反应 ,重组OviIL2在融合基因工程疫苗中具有良好的佐剂作用更多还原

【Abstract】 A fusion pili and ovine IL₂ subunit protein was expressed by transforming the plasmid pmPiliIL2 harbouring pili gene and ovine IL2 gene into the competent host cell BL₂1（DE3）.A vaccine called fusion pili_oviIL2 vaccine was prepared from sonicated cell mixtures of the transformants with pili_oviIL2 fusion protein.Two rabbits of 2 groups were respectively inoculated subcutaneously with either a single fusion pili_oviIL2 vaccine or an emulsion of pili_oviIL2 vaccine with Freund’s adjuvant,and each received the same dose at interval 3 weeks.Blood samples for test rabbits were taken for counter immunoelectrophoresis after inoculation at interval of one week to evaluate the antibody response.Three sheep in group 1 received two times of inoculation with either a single fusion pili_oviIL2 vaccine without any adjuvant and two sheep in group 2 received an emulsion of a recombinant pili vaccine with Freund’s adjuvant.Humoral antibody response was evaluated as did for rabbits above.Results showed that the serum antibody was elicited at 7th day after first inoculation and sustained over 6 months in each of four test rabbits inoculated with fusion pili_oviIL2 vaccines with or without Freund’s adjuvant.The same result was also observed in test sheep.The antibody response occurred in three sheep inoculated with a single pili_oviIL2 earlier at days 7 and 14 respectively while the antibody response for sheep inoculated with a recombinant pili vaccine was observed at 28th day. All of test sheep observed remained positive for antibody response over 6 months.Thus it can be derived that the single fusion pili_oviIL2 vaccine without any adjuvant elicited a high humoral immune response and the fusion oviIL₂ subunit in the vaccine had at least the same effective adjuvant activity in enhancing immunogenicity as Freund’s adjuvant did.更多还原