摘要
从猪带绦虫六钩蚴 c DNA文库中筛选目的基因并进行克隆表达 ,重组抗原用血清学方法和猪体免疫试验进行鉴定 ,筛选保护性抗原基因。将具有免疫保护作用的重组抗原纯化 ,并与免疫刺激复合物佐剂结合 ,制成猪囊虫病基因工程疫苗。对工程菌的培养发酵、重组抗原的下游纯化及疫苗的配制、疫苗的中间试制进行了研究 ,确定了一套比较完善的生产工艺。应用昆明小鼠建立了猪囊虫病的实验动物模型 ,应用该模型进行上述疫苗的免疫预防试验 ,免疫 1次和 2次的免疫保护率分别为 96.9%和 98.4 %。本动物的免疫预防试验显示 ,疫苗安全性好 ,免疫保护率达 92 .2 % ,且免疫组发现的囊虫多数已死亡。在流行区进行了田间试验和区域试验。免疫猪未有不良反应 ,囊虫感染率分别由 2 0 %降为 1 .1 %和 5.4 %降为 0 .2 1 %。用该疫苗对人工感染的囊虫病猪进行免疫治疗 ,发现在感染早期的治疗效果较好。免疫动物的体液免疫和细胞免疫的检测结果显示 ,该疫苗刺激机体产生抗体的时间早、持续时间长、效价高 ;可显著提高淋巴细胞转化率及 E-RFC和 Ea-RFC细胞、ANAE+细胞和粗粒型 ANAE+细胞、抑制 /杀伤性 T细胞亚群的数量。以上结果表明 ,用大肠杆菌表达的重组抗原制备的猪囊虫病基因工程疫苗安全、高效 ,且成本低廉 ,可规模化生产 ,
The purpose genes were screened from the cDNA library of Taenia solium oncospheres, subcloned and expressed. In order to screen the genes of the protective antigens , the expressed recombinant antigens were identified by serological method and the immunized pigs , respectively. The genetic engineering vaccine against swine cysticercosis was made from the purified protective antigen and the ISCOM adjuvant. The animal model of Kunming mouse for swine cysticercosis was established and used to carry out immunoprophylaxis against swine cysticercosis with the genetic engineering vaccine ,the immunological protective rates of one injection group and two injection group were 96.9% and 98.4%, respectively. Piglets were immunized with the vaccine and challenged orally by viable T. solum eggs. The vaccine was safety, no toxic and side effects to piglets, with the protective rate of 92.2% in contrast to the non vaccinated group. Most of the cysticerci found in the immunized group were dead. The effects of the immunotherapy for the genetic engineering vaccine were discovered , and the earlier the therapy the better the effect. The small scale and large scale field vaccination were carried out in the endemic area by immunizing 3583 and 20681 piglets, with the infection rates of cysticercitobe decreased from 20% to 1.1% and 5.4% to 0.21%, respectively. In order to probe the immune mechanism of the vaccine , the humoral and cellular immune response were detected. The detecting results of humoral immune revealed that some animals appeared antibody response against swine cysticercosis at 7 days after immunization,the Abs of all the immumized animals were positive at 21 d. The lasting time of the Ab was more than 210 d and Ab titrer was higher than the Al(oH)3 gel vaccine. The detecting results of the cellular immune displayed that lymphocyte transformation rate, ANAE + cells and bulky ANAE + cell,E RFC and Ea RFC of the immunized pigs were promoted obviously,and the suppressor/killer T ymphocytes were higher than groups of Al(oH)3 gel vaccine and contrast. The above results appeared that the genetic engineering vaccine against swine cysticercosis, which was made of the recombinamt antigen expressed in E. coli, was safety, efficiency,low price and scaled productivity ,it would be a new biological preparation for preventing swine cysticercosis.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2001年第1期49-53,共5页
Chinese Journal of Veterinary Science
关键词
猪囊虫病
基因工程疫苗
研制
免疫预防
免疫治疗
swine cysticercosis
genetic engineering vaccine
development
immunoprophylaxis
immunotherapy
