C群和W135群流脑结合疫苗的制备及其初步免疫评价

doi:10.12034/j.issn.1009-606X.218122

摘要/Abstract

摘要： 用NaIO4分别衍生C群和W135群脑膜炎球菌荚膜多糖(GCMP和GWMP)，经异端基双官能团交联剂N-β-马来酰亚胺丙酸酰肼偶联破伤风类毒素TT，制备两种多糖结合疫苗并免疫BALB/c小鼠，采用间接ELISA法测定血清中多糖特异性抗体和TT特异性抗体，采用硫氰酸盐洗脱法测定多糖特异性抗体亲合力. 结果表明，GCMP?TT和GWMP?TT结合物的多糖蛋白比分别约为0.4和1.0；初次免疫后两周、一次和二次加强免疫后各一周，GCMP?TT组的抗C糖特异性抗体滴度分别是GCMP组的3.2, 1.4和1.2倍，而GWMP?TT组则分别为GWMP组的2.5, 2.2和2.8倍，表明两种结合物均可有效提升纯抗原的免疫效果；两个结合物组的血清免疫球蛋白亚型的比值IgG2a/IgG1均高于多糖对照组，表明结合载体蛋白后多糖由TI抗原转变为TD抗原；两种结合物的多糖特异性抗体亲和力指数约为纯抗原的2倍，表明结合物均可诱导出较强的免疫记忆.

关键词: 流行性脑脊髓膜炎, 多糖, 异端基双官能团交联剂, 载体蛋白, 结合疫苗

Abstract: Bacterial meningitis is an acute infection of the fluid surrounding the brain and spinal cord that can rapidly lead to death. Capsular polysaccharides are a major virulence factor and form the basis for serogroup designation and protective vaccines. Polysaccharide vaccines have been replaced by glycoconjugate vaccines in order to improve the immunogenicity in infants and young children and inducing memory responses. The conjugates also induced high antibody avidity and prolong the IgG persistence. Moreover, compared with the polysaccharide vaccine, the conjugate vaccine can induce a long-lasting, protective memory T-cell response and elicit a comprehensive humoral and cellular immune response. In this study a new synthetic strategy was used to prepare two kinds of monovalent effective meningococcal vaccines. Group C and group W135 meningococcal capsular polysaccharide (GCMP and GWMP) were aldylated with sodium periodate, followed by the conjugation with tetanus toxoid (TT) via the heterotelechelic crosslinker N-β-maleimidopropionic acid hydrazide, separately. The BALB/c mice were immunized with the two conjugates. The polysaccharide (PS) specific antibody and the TT specific antibody in sera by indirect ELISA, and the avidity of PS specific antibody by thiocyanate elution were determined. The results showed that the mass ratios of PS to protein were 0.4 (GCMP-TT) and 1.0 (GWMP-TT). The PS specific antibody in GCMP-TT group were 3.2, 1.4 and 1.2 times, higher than those in GCMP group in primaty, 1st boost and 2nd boost, which were 2.5, 2.2 and 2.8 in GWMP-TT group, indicating the immune enhancement of both conjugate vaccines. The ratios of IgG2a to IgG1 were much higher in PS-TT groups than that in PS groups, demonstrated the conversion of TI antigen to TD type. The doubled avidity index of PS specific antibody in PS-TT groups showed the effective immune memory induced by the conjugation of carrier proteins to polysaccharides. This study confirmed that this synthetic route can effectively synthesize vaccines with better immunogenicity and improve the immune effect of the meningitis vaccine, and has a good application prospect.

Key words: Epidemic cerebrospinal meningitis, polysaccharide, heterotelechelic crosslinker, carrier protein, conjugate vaccine

江苗苗张竞鲁勖琳. C群和W135群流脑结合疫苗的制备及其初步免疫评价[J]. 过程工程学报, 2018, 18(6): 1283-1292.

Miaomiao JIANG Jing ZHANG Xulin LU. Preparation and preliminary immunoassay of group C and group W135 meningococcal polysaccharide conjugate vaccines[J]. Chin. J. Process Eng., 2018, 18(6): 1283-1292.

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