González S, Nazábal C, Viña L, Caballero E
Center for Genetic Engineering and Biotechnology, Havana, Cuba.
Dev Biol Stand. 1998;92:269-76.
Studying outer membrane proteins as vaccine candidates, our group has previously isolated, cloned, and expressed in Escherichia coli the gene encoding for a high molecular weight protein (P64k), common to many meningococcal strains. To continue the characterisation of this meningococcal antigen, we have evaluated its immunogenicity in mice alone or combined with several commercially-available adjuvants. We used as an adjuvant aluminium hydroxide (Alhydrogel and Rehydragel), aluminium phosphate, Algammulin, crude saponin, the saponin Quil A, dimethyl-dioctadecyl ammonium bromide (DDA), Freund's adjuvant, and Montanide 888. The antibody titres against the recombinant protein and whole meningococci elicited with these adjuvants were compared. We found that Quil A produced the highest titres against the recombinant P64k. Algammulin and the quaternary ammonium compound DDA induced the highest levels of antibodies against meningococci. We analysed the recognition of a set of linear peptides by antisera prepared against the protein combined with some of the adjuvants. The responses depended on the adjuvant used and the results have been confirmed by epitope mapping using overlapping peptides synthesised on pins.
作为疫苗候选物研究外膜蛋白时，我们小组先前已在大肠杆菌中分离、克隆并表达了编码许多脑膜炎球菌菌株共有的一种高分子量蛋白（P64k）的基因。为继续对这种脑膜炎球菌抗原进行特性分析，我们评估了其单独或与几种市售佐剂联合使用时在小鼠体内的免疫原性。我们使用的佐剂有氢氧化铝（Alhydrogel和Rehydragel）、磷酸铝、Algammulin、粗皂苷、皂苷Quil A、二甲基二十八烷基溴化铵（DDA）、弗氏佐剂和Montanide 888。比较了这些佐剂诱导产生的针对重组蛋白和完整脑膜炎球菌的抗体效价。我们发现Quil A产生的针对重组P64k的效价最高。Algammulin和季铵化合物DDA诱导产生的针对脑膜炎球菌的抗体水平最高。我们分析了用与某些佐剂联合使用的该蛋白制备的抗血清对一组线性肽的识别情况。反应取决于所用佐剂，并且通过使用在针上合成的重叠肽进行表位作图证实了结果。
