This invention discloses methods for identifying
Francisella tularensis
vaccine candidates. It enables identification of novel vaccine candidates and quality assurance for vaccine batches, assessment of protection in vaccinates and identification of the infecting agent in vaccinates. Mice were first vaccinated with
Brucella abortus
O-polysaccharide (OPS) vaccine. These animals were then given 10 LD
50
s of
F. tularensis
live vaccine strain (LVS). Sixty percent (60%) of the vaccinated mice survived the multiple lethal doses. Sera were collected from these surviving mice and the antibodies were used to probe supernatant and cell lysates of live
F. tularensis
LVS cultures. Several
F. tularensis
components were identified only by the noted “survivor” antisera. Of these identified proteins, enzyme digestions and chemical oxidation suggest post-translational modifications of some proteins e.g. a 52 kDa glycoprotein, a 45 kDa lipoprotein and a 19 kDa nucleoprotein. The 52 kDa component caused nitrous oxide induction in tissue cultures at low concentrations, cell death at high concentrations. Vaccination with this gave partial protection while addition of other components acted synergistically to give enhanced protection from 250 LD
50
s of
F. tularensis
LVS.
本发明公开了用于鉴定
土拉弗氏菌
候选疫苗的方法。通过该方法可以鉴定新型候选疫苗,保证疫苗批次的质量,评估疫苗的保护作用,鉴定疫苗的感染源。首先用
流产布鲁氏菌
O-
多糖(OPS)疫苗。然后给这些动物注射 10 LD
50
的
土拉菌
活疫苗菌株(LVS)。60%的接种小鼠在多次致死剂量后存活下来。从这些存活的小鼠身上收集血清,用
抗体探查活的
F. tularensis
LVS 培养物。几种
土拉菌
只有 "幸存者 "抗血清能鉴定出其中的几种成分。在这些已鉴定的蛋白质中,酶消化和
化学氧化表明一些蛋白质发生了翻译后修饰,如 52 kDa 糖蛋白、45 kDa 脂蛋白和 19 kDa 核蛋白。52 kDa 成分在低浓度下会在组织培养物中引起
一氧化二氮,在高浓度下会导致
细胞死亡。接种该成分的疫苗可提供部分保护,而添加其他成分则可发挥协同作用,在 250 LD
50
s 的
F. tularensis
LVS。