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目的利用不同的载体对脑膜炎奈瑟球菌(Neisseria meningitidis,Nm)A群、B群、C群、W135群和Y群进行偶联,比较不同的免疫组合物的免疫效果。方法将Nm A群、W135群和Y群多糖与CRM197偶联;将B群Nm菌种进行传代、培养、提取并纯化外膜蛋白,将外膜蛋白与C群多糖偶联。采用冻干水化法将蛋白质-多糖偶联物与脂质体(以明矾作为对照)混合后,免疫豚鼠,对不同Nm免疫组合物的杀菌滴度进行检测。结果各免疫组合物针对相应的Nm血清群具有不同的免疫作用,与C群多糖和B群外膜蛋白直接混合后免疫相比,C群多糖与B群外膜蛋白以共价键偶联后形成的免疫复合物免疫豚鼠,可引发更高滴度的杀菌抗体;而与以明矾作为佐剂相比,以脂质体作为佐剂均具有更好的免疫促进作用,其中以A群和Y群的免疫增强效果明显(P<0.05)。结论以不同抗原共价键偶联的方式较抗原直接混合后制备的Nm免疫组合物具有更高的杀菌滴度,同时以脂质体作为佐剂有利于增强脑膜炎疫苗的免疫效果。
OBJECTIVE: To compare the immune effects of different immune compositions by using different vectors to couple Neisseria meningitidis group A, B group, C group, W135 group and Y group. Methods Nm A, W135 and Y polysaccharides were conjugated with CRM197. The Nm strains of B group were subcultured, cultured, and the outer membrane protein was extracted and purified. The outer membrane protein was coupled with C group polysaccharides. After the protein-polysaccharide conjugate was mixed with liposomes (with alum as a control) by freeze-drying hydration method, the guinea pigs were immunized and the bactericidal titers of different Nm immune compositions were tested. Results The results showed that each immune composition had different immunological effects against the corresponding Nm serogroups. Compared with the immunization of C group polysaccharides and B group outer membrane proteins, C group polysaccharides were covalently coupled with B group outer membrane proteins The immunocomplex formed immunized guinea pigs, can lead to higher titers of bactericidal antibodies; and compared with alum as an adjuvant, with liposomes as adjuvants have a better immune boost, which group A and Y The immune enhancement group was significantly (P <0.05). Conclusion Nm immunosuppressant prepared by direct covalent coupling of different antigens has higher bactericidal titer than liposome, and the liposome as an adjuvant is beneficial to enhance the immunogenicity of meningitis vaccine.