β-丙内酯在Vero细胞HFRS疫苗中的应用  

Application of β-propiolactone in HFRS vaccine

作　　者：陈伟[1] 李忠义[2] 刘江秋[2] 于笑难[3] 薛采芳[1] 蔡群[4] 徐璐 廖辉 

机构地区：[1]第四军医大学基础部病原生物学教研室,西安710032 [2]沈阳军区军事医学研究所 [3]沈阳军区总医院检验科 [4]黑龙江省哈尔滨市第一医院 [5]辽宁卫星生物制品研究所

出　　处：《中国公共卫生》2003年第6期669-670,共2页Chinese Journal of Public Health

摘　　要：目的　探讨 β -丙内酯灭活双价Vero细胞肾综合征出血热纯化疫苗的最佳条件。 方法　采用不同终浓度的 β-丙内酯和不同的灭活时间对疫苗进行灭活 ,用细胞培养法进行病毒增殖试验 ,以确定灭活效果。应用高效液相色谱法分析 β -丙内酯的最佳水解条件和 3批双价vero细胞出血热纯化灭活疫苗 β-丙内酯残留量。检测 3批双价纯化灭活疫苗的免疫效果。结果　疫苗经终浓度为 1∶2 0 0 0和 1∶4 0 0 0的 β -丙内酯作用 2 4h ,均可完全灭活病毒 ;疫苗中的 β-丙内酯在 37℃水浴下 ,随着水解时间的延长 ,其含量逐渐下降 ,水解 2h后已无 β-丙内酯检出 ;3批经 β-丙内酯灭活的双价Vero细胞出血热纯化疫苗均未检出 β-丙内酯残留 ,在家兔体内诱导产生针对汉滩型和汉城型病毒的中和抗体效价均达到 1∶2 0。结论　肾综合征出血热纯化疫苗经终浓度为 1∶4 0 0 0 β -丙内酯 4℃灭活 2 4h ,然后再于 37℃水浴中水解 2h ,既可达到完全灭活病毒且无 β-丙内酯残留的目的 ;Objective To study the optimal inactivated condition of β-propiolactone(BPL) on preparation of hemorrhagic fever with renal syndrome(HFRS)bivalent purified vaccine(vero cells derived).Methods The HFRS bivalent purified vaccine was treated with different final concentrations and different time respectively.Residual infectivity was determimed by standard viral proliferation assay on tissue culture cells.Using high performance liquid chromatography(HPLC),the optimal condition of hydrolyssi of bpl was analyzed and the residual contents of BPL in 3 batches of HFRS bivalent purified and inactivated vaccine were measured.Rabbits immunized with the 3 batches of HFRS bivalent purified and inactivated vaccine were detected the neutralizing antibody titers of sera at 4 weeks after 2 times of immunization.Results The HFRS bivalent purified vaccines treated with the final concentration of 1∶2?000 and 1∶4?000 of BPL for 24 hours were inactivated completely.Under the condition of 37?℃ water bath,the content of BPL in HFRS vaccine decreased gradually along with the prolongation of time and disappeared completely 2 hous later.Rabbits immunized with the 3 batches of HFRS bivalent purified and inactivated vaccine showed 100% sera conversion and the neutralizing antibody titers of sera against hantaan virus(HTN) and seoul virus(SEO) reached 1∶20.Conclusion It was feasible and secure for inactivation of HFRS bivalent purified vaccine(vero cells derived)with the final concentration of 1∶4?000 of BPL under the conditions of 4?℃ for 24 hours and then 37?℃ water bathing for 2 hours.The HFRS bivalent purified vaccine inactivated with BPL possessed good immunogenicity.

关 键 词：肾综合征出血热 疫苗 VERO细胞 β-丙内酯 病毒灭活 高效液相色谱 

分 类 号：R373.32[医药卫生—病原生物学]