Heteropentameric cholera toxin B subunit chimeric molecules genetically fused to a vaccine antigen induce systemic and mucosal immune responses: a potential new strategy to target recombinant vaccine antigens to mucosal immune systems
Heteropentameric cholera toxin B subunit chimeric molecules genetically fused to a vaccine antigen induce systemic and mucosal immune responses: a potential new strategy to target recombinant vaccine antigens to mucosal immune systems
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DOI：
10.1128/iai.73.9.5654-5665.2005
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发表时间：
2005-09-01
影响因子：
3.1
通讯作者：
Arakawa, T
中科院分区：
文献类型：
--
作者：
Harakuni, T;Sugawa, H;Arakawa, T
非侵入性粘膜疫苗是胃肠外疫苗的有吸引力的替代品。尽管疫苗抗原与霍乱毒素的B亚基（CT B）的缀合是用于疫苗递送至粘膜免疫系统的最有前途的策略之一，但是该分子不能耐受大蛋白融合，因为它严重损害五聚体化并且失去对GM 1-神经节苷脂的亲和力。在这里，我们报告了一种新的策略，其中CTB-抗原融合亚基之间的空间位阻通过将未融合的CTB“分子缓冲液”整合到五聚体单元中而显著降低，使它们更有效地自组装成具有生物活性的五聚体。此外，嵌合蛋白采取了紧凑的配置，变得足够小的分泌，和一步亲和纯化的蛋白质，当通过粘膜途径给药时，诱导小鼠的特异性免疫应答。由于我们的结果不依赖于特定表达系统或疫苗抗原的使用，因此该策略可广泛适用于基于细菌肠毒素的疫苗设计。
Noninvasive mucosal vaccines are attractive alternatives to parenteral vaccines. Although the conjugation of vaccine antigens with the B subunit of cholera toxin (CTB) is one of the most promising strategies for vaccine delivery to mucosal immune systems, the molecule cannot tolerate large-protein fusion, as it severely impairs pentamerization and loses affinity for GMI-ganglioside. Here we report a new strategy, in which steric hindrance between CTB-antigen fusion subunits is significantly reduced through the integration of unfused CTB "molecular buffers" into the pentamer unit, making them more efficiently self-assemble into biologically active pentamers. In addition, the chimeric protein took a compact configuration, becoming small enough to be secreted, and one-step affinity-purified proteins, when administered through a mucosal route, induced specific immune responses in mice. Since our results are not dependent on the use of a particular expression system or vaccine antigen, this strategy could be broadly applicable to bacterial enterotoxin -based vaccine design.