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作者用含表达B群脑膜炎球菌外膜蛋白P1的基因质粒的枯草杆菌生产出重组蛋白BacP1,经多步提纯及加热变性后,用磷脂酰胆碱把BacP1重建于脂质体中,形成天然抗原表位,制成BacP1-脂质体菌苗。分别于0和42天皮下注射NIH小鼠,于第52天采血,用酶免疫测定法(EIA)检测抗体滴度。结果显示,小鼠产生了高EIA滴度的抗同源变性和天然型抗原的抗体,且抗体滴度随免疫剂量的增加而升高,而产生的抗异源变性和天然型抗原的抗体滴度则很低。用脑膜炎球菌株作靶抗原检测血清的杀菌能力显示,血清的杀菌活性与EIA滴度一致,1～3μg免疫量产生的杀菌抗体滴度为16～32,6～10μg免疫量产生的杀菌抗体滴度为64～128,且杀菌活性是P1亚型特异性的。用幼大鼠脑膜炎模型试验血清的保护效力显示,注射免疫量为6～10μg的小鼠血清的大鼠,经同型菌株攻击后,完全获得保护,注射免疫量为1～3μg的小鼠血清的大鼠,仅部分大鼠获得保护,而注射未免疫小鼠血清的大鼠,经同样菌株攻击后,全部发生脑膜炎。作者还用此菌苗免疫了Balb/c小鼠和豚鼠,结果均获得较高的抗体水平,个体差异很小。
The authors used a Bacillus subtilis containing a plasmid expressing the meningococcal outer membrane protein P1 to produce the recombinant protein BacP1. After several steps of purification and heat denaturation, BacP1 was reconstructed into liposomes with phosphatidylcholine to form a natural Antigen epitope, made BacP1-liposome vaccine. NIH mice were injected subcutaneously on days 0 and 42, respectively, and blood was collected on day 52 for detection of antibody titers by enzyme immunoassay (EIA). The results showed that mice produced high EIA titers of antibodies against homologous degeneration and native antigens and that the antibody titers increased with increasing immunization dose and that the resultant antibody titers against heterologous degenerations and native antigens Degree is very low. The bactericidal activity of the meningococcal serogroups as the target antigen for the detection of serum showed that the bactericidal activity of the serum was consistent with the titer of the EIA, and the bactericidal antibody titers produced by the immunized amount of 1 to 3 μg were 16 to 32 and the immunogenicity of 10 to 10 μg The titers ranged from 64 to 128 and the bactericidal activity was specific for the P1 subtype. The protective efficacy of the serum from the rat meningitis model test showed that the mice immunized with the mouse serum with an immunization dose of 6 to 10 μg were fully protected after being challenged by the same type strain and the mice were immunized with the immunized amount of 1-3 μg Of rats, only part of the rats were protected, while rats injected with non-immunized mice sera all developed meningitis after challenge with the same strain. The authors also vaccinated Balb / c mice and guinea pigs with this bacterin, resulting in higher antibody levels with little individual variation.