A non-toxic recombinant protein rSUMO-CPBm4 as a potential vaccine candidate against Clostridium perfringens type C beta enterotoxemia.

Tropical biomedicine Pub Date : 2023-12-01 DOI:10.47665/tb.40.4.004

Y Goa, J G Du, C Jirapattharasate, E Galon, S W Ji, Z G Ran, Y Q Xia

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Abstract

Beta toxin (CPB) is a lethal toxin and plays a key role in enterotoxemia of ruminants caused by Clostridium perfringens type C strain. The existing vaccines based on crude CPB need time-consuming detoxification and difficult quality control steps. In this study, we synthesized the rCPB_m4 of C. perfringens type C strain and small ubiquitin-like modifier (SUMO)-tag CPB_m4 (rSUMO-CPB_m4) by introducing four amino acid substitutions: R212E, Y266A, L268G, and W275A. Compared with rCPB_m4, rSUMO-CPB_m4 was expressed with higher solubility in Escherichia coli BL21 (DE3). Neither rCPB_m4 nor rSUMO-CPB_m4 was lethal to mice. Although rCPB_m4 and rSUMO-CPB_m4 were reactogenic with polyclonal antibodies against crude CPB, rabbits vaccinated with rSUMO-CPB_m4 developed significant levels of toxin-neutralizing antibody (TNA) titers that conferred protection against crude toxin challenge. These data suggest that genetically detoxified rSUMO-CPB_m4 is a promising subunit vaccine candidate for C. perfringens type C beta enterotoxemia.

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一种无毒重组蛋白 rSUMO-CPBm4 可作为预防 C 型产气荚膜梭菌 beta 肠毒血症的候选疫苗。

贝塔毒素（CPB）是一种致命毒素，在 C 型产气荚膜梭菌引起的反刍动物肠毒血症中起着关键作用。现有的基于粗制 CPB 的疫苗需要耗时的解毒和困难的质量控制步骤。在本研究中，我们通过引入四个氨基酸取代，合成了 C 型产气荚膜梭菌的 rCPBm4 和小泛素样修饰物（SUMO）标签 CPBm4（rSUMO-CPBm4）：R212E、Y266A、L268G 和 W275A。与 rCPBm4 相比，rSUMO-CPBm4 在大肠杆菌 BL21 (DE3) 中的表达溶解度更高。rCPBm4 和 rSUMO-CPBm4 都不会使小鼠致死。虽然 rCPBm4 和 rSUMO-CPBm4 与针对粗制 CPB 的多克隆抗体有反应性，但接种了 rSUMO-CPBm4 的兔子体内产生了大量毒素中和抗体 (TNA)，这些抗体能保护兔子免受粗制毒素的挑战。这些数据表明，基因解毒的 rSUMO-CPBm4 是一种很有前景的候选亚单位疫苗，可用于治疗 C 型产气荚膜杆菌肠毒血症。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Tropical biomedicine

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