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Abstract：
There is a pressing need for new vaccines against alphaviruses, which can cause fatal encephalitis (Venezuelan equine encephalitis virus (VEEV) and others) and severe arthralgia (e.g. Chikungunya virus, CHIKV). These positive-strand RNA viruses are diverse and evolve rapidly, meaning that the sequence of any vaccine should cover multiple strains that may be quite different from any previous isolate. Here, consensus proteins were produced to represent the common physicochemical properties (PCPs) of the epitope rich, B domain of the E2 envelope protein. PCP-consensus proteins were based on multiple strains of VEEV (VEEVcon) and CHIKV (CHIKVcon) or the conserved PCPs of 24 different alphaviruses (AllAVcon). The AllAVcon was altered to include binding sites for neutralizing antibodies of both VEEV and CHIKV strains (Mosaikcon). All four designed proteins were produced solubly in E. coli and purified. They formed the β-strand core expected from experimental structures of this region of the wild type E2 proteins as indicated by circular dichroism (CD) spectra. Furthermore, the CHIKVcon protein bound to a structure dependent, CHIKV neutralizing monoclonal antibody. The AllAVcon and Mosaikcon proteins bound to polyclonal antibodies generated during natural infection with either VEEV or CHIKV, indicating they contained epitopes of both serotypes. The Mosaikcon antigen induced antibodies in rabbit sera that recognized both the VEEVcon and CHIKVcon spike proteins. These PCP-consensus antigens are promising starting points for novel, broad-spectrum alphavirus vaccines.
摘要：
迫切需要针对甲病毒的新疫苗，这可能导致致命的脑炎（委内瑞拉马脑炎病毒（VEEV）和其他）和严重的关节痛（例如基孔肯雅病毒，CHIKV）。这些正链RNA病毒种类繁多，进化迅速，这意味着任何疫苗的序列都应涵盖多种菌株，这些菌株可能与以前的任何分离株完全不同。这里,产生的共有蛋白代表富含表位的共同物理化学性质(PCPs),E2包膜蛋白的B结构域。PCP共有蛋白基于VEEV（VEEVcon）和CHIKV（CHIKVcon）的多种菌株或24种不同甲病毒（AllAVcon）的保守PCP。改变AllAVcon以包括用于中和VEEV和CHIKV毒株(Mosaikcon)的抗体的结合位点。所有四种设计的蛋白质在大肠杆菌中可溶地产生并纯化。它们形成了由野生型E2蛋白的该区域的实验结构所预期的β-链核心,如通过圆二色性(CD)光谱所指示的。此外，CHIKVcon蛋白与结构依赖结合，CHIKV中和单克隆抗体。AllAVcon和Mosaikcon蛋白与VEEV或CHIKV自然感染过程中产生的多克隆抗体结合，表明它们含有两种血清型的表位。Mosaikcon抗原在兔血清中诱导识别VEEVcon和CHIKVcon刺突蛋白的抗体。这些PCP共有抗原是有希望的新的起点，广谱甲病毒疫苗。
