无毒性逆转的无细胞百日咳菌苗制备

Preparation of Irreversible Toxoiding of Aceiiular Pertussis Vaccine

将百日咳菌培养物上清液,经一段硫酸铵盐析提取百日咳主要保护性抗原 FHA 和 PT,用蔗糖密度梯度离心去除内毒素,以戊二醛解毒制成无细胞百日咳菌苗。该菌苗对家兔不产生热原反应,对小鼠毒性低,不引起明显的白细胞增多及组胺致敏活性,小鼠免疫后对百日咳毒菌脑腔攻击具有很好的保护作用,并可产生较高的抗体应答。菌苗经37℃保存3个月未出现毒性逆转现象,质量稳定,为大批量制造质量均一的无细胞百日咳菌苗提供了可能性。

B.Pertussis phase I was cultivated in SS medium supplemented with 0.1%Heptakis (dimethyl-b-cyclodextrin) in large shaking bottoles.From the culture superna-tant,two kinds of main protective antigens FHA and PT were extracted by precipita-tion with ammonium sulfate and endotoxin was removed by density gradient centrifug-ation.The FHA and PT preparation was detoxified in an appropriate degree with glu-taraldehyde.Finally the acellular pertussis vaccine (ADPT) was obtained.The test showed that ADPT vaccine did not cause the fever in rabbits and ind-uce leucocytosis and histamin sensitizing activity inmice significantly.The vaccineprepared by the method mention-above conferrd good protection and higher antib-ody response to B.pertussis in mice.Toxicity reversion of the vaccine was not fo-und when stored at 37℃ for 3 months.