Immunization with a novel Clostridium perfringens epsilon toxin mutant rETX(Y196E)-C confers strong protection in mice.
Immunization with a novel Clostridium perfringens epsilon toxin mutant rETX(Y196E)-C confers strong protection in mice.
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DOI：
10.1038/srep24162
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发表时间：
2016-04-06
影响因子：
4.6
通讯作者：
Wang J
中科院分区：
文献类型：
--
作者：
Yao W;Kang J;Kang L;Gao S;Yang H;Ji B;Li P;Liu J;Xin W;Wang J
Epsilon 毒素 (ETX) 由产气荚膜梭菌毒素 B 型和 D 型产生。它可诱发家畜（主要是绵羊、山羊和牛）致命的肠毒血症，给全球畜牧业造成严重的经济损失。在这项研究中，通过用谷氨酸（E）取代第196位酪氨酸（Y196）并引入23个氨基酸长的C端肽而获得的新型稳定的ε毒素突变体rETXY196E-C被确定为有前途的抗肠毒血症重组疫苗候选物。第三次接种后，各免疫组抗重组野生型（rETX）的抗体滴度可达1:105，小鼠完全免受100 × LD50（50%致死量）rETX攻击。 15μg皮下免疫组的小鼠在rETX攻击剂量为500 × LD50时完全存活，而在180μg（1000 × LD50）剂量的rETX给药时80％的小鼠存活。在体外，15μg皮下免疫组的免疫血清可以完全保护MDCK细胞免受16× CT50（50％细胞致死剂量）rETX攻击的影响，并能保护小鼠免受10× LD50剂量（1.8μg）rETX攻击的影响。这些数据表明重组蛋白 rETXY196E-C 是未来应用研究的潜在候选疫苗。
Epsilon toxin (ETX) is produced by toxinotypes B and D of Clostridium perfringens. It can induce lethal enterotoxemia in domestic animals, mainly in sheep, goats and cattle, causing serious economic losses to global animal husbandry. In this study, a novel and stable epsilon toxin mutant rETXY196E-C, obtained by substituting the 196th tyrosine (Y196) with glutamic acid (E) and introducing of 23 amino acids long C-terminal peptide, was determined as a promising recombinant vaccine candidate against enterotoxemia. After the third vaccination, the antibody titers against recombinant wild type (rETX) could reach 1:105 in each immunized group, and the mice were completely protected from 100 × LD50 (50% lethal dose) of rETX challenge. The mice in 15 μg subcutaneously immunized group fully survived at the dose of 500 × LD50 of rETX challenge and 80% of mice survived at 180 μg (1000 × LD50) of rETX administration. In vitro, immune sera from 15 μg subcutaneously immunized group could completely protect MDCK cells from 16 × CT50 (50% lethal dose of cells) of rETX challenge and protect against 10 × LD50 dose (1.8 μg) of rETX challenge in mice. These data suggest that recombinant protein rETXY196E-C is a potential vaccine candidate for future applied researches.