CAJ | 학술논문

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BL-21，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，21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21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28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。
장전화절류의간균(D.nodosus)섬모단백(Pili)화면양백세포개소2(oviIL2)융합기인표체질립적공정균BL-21，재함사변청매소영양육탕배양기중표체，리심획득균체침정물，렬해후배제성Pili-oviIL2융합기인공정역묘。용가좌제화불가좌제적Pili-oviIL2융합기인공정역묘분별접충2지건강가토，21천후접충제2차；정기채혈，용대류면역전영검측시험토적체액면역반응。결과발현，가좌제화불가좌제적Pili-oviIL2융합기인공정역묘면역토7천즉가산생상응항체，항체재면역혈청중가유지6개월이상。진일보시험장불가좌제적Pili-oviIL2융합기인공정역묘접충3지건강면양，동양21천후접충제2차，정기채혈，용대류면역전영검측시험면양적체액면역반응。동시용Pili기인공정역묘접충2지면양작대조。결과3지면양접충Pili-oviIL2융합기인공정역묘후，분별우7천화14천산생상응적항체，이접충Pili기인공정역묘적면양우28천산생상응적항체；피면역면양혈청중적항체가유지6개월이상。용Pili-oviIL2융합기인공정역묘접충토화면양적면역시험표명，Pili-oviIL2융합기인공정역묘구유교호적체액면역응답반응，중조OviIL2재융합기인공정역묘중구유량호적좌제작용。
A fusion pili and ovine IL-2 subunit protein was expressed by transforming the plasmid pmPiliIL2 harbouring pili gene and ovine IL2 gene into the competent host cell BL-21(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 oviIL-2 subunit in the vaccine had at least the same effective adjuvant activity in enhancing immunogenicity as Freund's adjuvant did.