腐蹄病C型节瘤拟杆菌纤毛蛋白基因工程疫苗对家兔的免疫试验

Study on the Immunogenicity of the Recombinant Pili Protein Vaccine in Healthy Rabbits

将转化 C型节瘤拟杆菌纤毛蛋白基因表达子粒的工程菌 PAK/ 2 pfs C在含羧苄青霉素营养肉汤培养基中培养 ,用 Mg Cl2 法在培养物上清中提取表达产物 (纤毛蛋白 )。将表达的纤毛蛋白产物用弗氏完全佐剂乳化 ,制成疫苗。用疫苗免疫 3只健康家兔 ,2 1 d后再次接种 ;定期采血 ,用对流免疫电泳和 K凝集实验检测试验家兔的体液免疫应答及抗体水平。结果发现 ,免疫 7d即可产生相应抗体 ,2 1 d后抗体效价达到 80 0 0×以上 ,而且高滴度抗体可维持 6个月以上。试验表明 ,腐蹄病

The PME pili was transformed into the host competent cell PAK/2pfs and the recombinant pili was expressed in the supernatant of the cultures of the transformant cell PAK/2pfsC . the recombinant pili was purified by MgCl 2 from the supernatant of the culture of the transformed PAK/2pfs. Healty rabbit were inoculated vaccine of the recombinant pili protein at 250 μg per dose to potentiate the immunogenicity. Rabbit inoculated two time at interval 3 weeks. The blood of each rabbit was collected at days 0, 7, 14, 21, 30, 60, 90, 120, 150, 180. Blood samples for test rabbits were taken for counter immunoelectrophoresis. The antibody titers were evaluated by k agglutination. The results showed that the lower agglutination titers were observed at day 7 and higher levels over 8000 at day 21 induced by the vaccine of recombinant pili.All of test rabbits observed remained positive for antibody response of higher level over 6 months.