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陈伟, 李忠义, 刘江秋, 于笑难, 薛采芳, 蔡群, 徐璐, 廖辉. β-丙内酯在Vero细胞HFRS疫苗中的应用[J]. 中国公共卫生, 2003, 19(6): 669-670.
引用本文: 陈伟, 李忠义, 刘江秋, 于笑难, 薛采芳, 蔡群, 徐璐, 廖辉. β-丙内酯在Vero细胞HFRS疫苗中的应用[J]. 中国公共卫生, 2003, 19(6): 669-670.
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CHEN Wei, LI Zhong-yi, LIU Jiang-qiu, . Application of β-propiolactone in HFRS vaccine[J]. Chinese Journal of Public Health, 2003, 19(6): 669-670.
Citation: CHEN Wei, LI Zhong-yi, LIU Jiang-qiu, . Application of β-propiolactone in HFRS vaccine[J]. Chinese Journal of Public Health, 2003, 19(6): 669-670.
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β-丙内酯在Vero细胞HFRS疫苗中的应用

Application of β-propiolactone in HFRS vaccine

    摘要
  • 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1:2000和1:4000的β-丙内酯作用24h, 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1:20。
      结论   肾综合征出血热纯化疫苗经终浓度为1:4000β-丙内酯4℃灭活24h, 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; 采用此方法灭活的双价Vero细胞肾综合征出血热纯化疫苗保持良好的免疫原性。

     

    Abstract:
      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 prolonga2 tion 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 againsthantaan 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.

     

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