Tricomponent fusion complex comprising a viral antigen, a pentameric alpha-helical coiled-coil, and an immunoglobulin-binding domain as an effective antiviral vaccine.
Tricomponent fusion complex comprising a viral antigen, a pentameric alpha-helical coiled-coil, and an immunoglobulin-binding domain as an effective antiviral vaccine.
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三组分融合复合物包含病毒抗原、五聚体α螺旋卷曲螺旋和免疫球蛋白结合结构域，作为有效的抗病毒疫苗。
DOI：
10.1016/j.vaccine.2013.12.016
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发表时间：
2014
期刊：
影响因子：
5.5
通讯作者：
Tadano M
中科院分区：
文献类型：
--
作者：
Arakawa T;Harakuni T;Miyata T;Tafuku S;Tadano M
将软骨寡聚基质蛋白（COMP）的五聚体卷曲螺旋结构域与金黄色葡萄球菌蛋白A的Z结构域（一种免疫球蛋白结合结构域（IBD））基因融合，作为病毒抗原载体复合物进行评价。在概念验证研究中，通过化学缀合将重组日本脑炎病毒（JEV）E蛋白结构域III（D3）加载到COMP-Z融合蛋白上，并在小鼠JEV感染模型中评价所产生的三组分复合物COMP-Z/D3的疫苗效力。免疫与复合物赋予更大的保护，对致命的JEV感染比卸载抗原。接下来，工程化三组分复合物，其中三个分子实体（D3抗原、COMP卷曲螺旋结构域和Z结构域）串联遗传连接以产生D3-COMP-Z三组分复合物或其反向定向构建体Z-COMP-D3。融合蛋白在大肠杆菌中以包涵体形式表达，但可复性为具有生物活性的五聚体，保留了E蛋白的抗原性和IBD功能。免疫与重折叠复合物赋予了高水平的保护，对致命的JEV感染，类似的功效的三组分复合物产生的化学共轭。这些结果表明，三组分复合物，无论是通过化学或基因融合产生的，是一个有前途的分子设计，用于创建有效的亚单位疫苗抗病毒感染。
The pentameric coiled-coil domain of cartilage oligomeric matrix protein (COMP) genetically fused to the Z domain ofStaphylococcus aureusprotein A, an immunoglobulin-binding domain (IBD), was evaluated as a viral antigen carrier complex. In a proof-of-concept study, recombinant Japanese encephalitis virus (JEV) E protein domain III (D3) was loaded onto the COMP–Z fusion protein by chemical conjugation, and the tricomponent complex generated, COMP–Z/D3, was evaluated for its vaccine efficacy in a mouse JEV infection model. Immunization with the complex conferred substantially greater protection against lethal JEV infection than the unloaded antigen. Next, a tricomponent complex was engineered in which the three molecular entities (the D3 antigen, COMP coiled-coil domain, and Z domain) were genetically connected in tandem to create the D3–COMP–Z tricomponent complex, or its reversal oriented construct, Z–COMP–D3. The fusion complexes were produced as inclusion bodies inEscherichia coli, but could be refolded to biologically active pentamers that retained the E protein antigenicity and the IBD function. Immunization with the refolded complexes conferred a high level of protection against lethal JEV infection, similar in efficacy to that of the tricomponent complex generated by chemical conjugation. These results demonstrate that the tricomponent complex, whether generated by chemical or genetic fusion, is a promising molecular design for the creation of effective subunit vaccines against viral infections.