{"title":"2型志贺毒素e变体(Stx2e)的B亚基与五链α-螺旋线圈捆绑在一起,可以保护仔猪免受猪水肿病的侵害","authors":"Takeshi Arakawa , Hirotaka Uefuji , Yukihiro Tamaki , Shigeki Oogai , Hikaru Arakawa","doi":"10.1016/j.vaccine.2025.127140","DOIUrl":null,"url":null,"abstract":"<div><div>Porcine edema disease (ED) is caused by infection with Shiga toxin type 2 e variant (Stx2e)-producing <em>Escherichia coli</em> (STEC). To develop a new ED vaccine, we engineered a fusion protein, in which the Stx2e B subunit (Stx2eB) was fused to the five-stranded α-helical coiled coil domain of cartilage oligomeric matrix protein (COMP), based on our recent finding that the coiled coil strongly stabilizes the B subunit pentamer of Stx2 (<strong>Tamaki Y, Harakuni T, Arakawa T. Shiga toxin type 2 B subunit protects mice against toxin challenge when leashed and bundled by a stable pentameric coiled-coil molecule. Vaccine. 2024 Mar 7;42(7):1757–1767</strong>). Purified Stx2eB–COMP fusion protein administered to mice conferred complete protection against a lethal dose of Stx2e. However, unfused Stx2eB conferred only 10 % protection. Furthermore, when Stx2eB was fused to a trimeric or tetrameric coiled coil, a marked reduction in protective efficacy was observed, indicating the importance of “five-to-five” fusion stoichiometry. Next, we immunized weaned piglets twice with 100 μg or 10 μg of the Stx2eB–COMP fusion protein, and then orally challenged the animals with a lethal dose of STEC. The piglets immunized with the high dose were almost completely free from clinical symptoms of ED, whereas three of the six piglets administered adjuvant-only died, and the remaining surviving piglets exhibited severe ED symptoms. Although all piglets immunized with the low dose survived, they exhibited mild to moderate ED symptoms. Our findings indicate that Stx2eB is highly protective only when bundled and molecularly stabilized by the coiled coil molecule.</div></div>","PeriodicalId":23491,"journal":{"name":"Vaccine","volume":"61 ","pages":"Article 127140"},"PeriodicalIF":4.5000,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"B subunit of the type 2 Shiga toxin e variant (Stx2e) bundled by a five-stranded α-helical coiled coil protects piglets from porcine edema disease\",\"authors\":\"Takeshi Arakawa , Hirotaka Uefuji , Yukihiro Tamaki , Shigeki Oogai , Hikaru Arakawa\",\"doi\":\"10.1016/j.vaccine.2025.127140\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div>Porcine edema disease (ED) is caused by infection with Shiga toxin type 2 e variant (Stx2e)-producing <em>Escherichia coli</em> (STEC). To develop a new ED vaccine, we engineered a fusion protein, in which the Stx2e B subunit (Stx2eB) was fused to the five-stranded α-helical coiled coil domain of cartilage oligomeric matrix protein (COMP), based on our recent finding that the coiled coil strongly stabilizes the B subunit pentamer of Stx2 (<strong>Tamaki Y, Harakuni T, Arakawa T. Shiga toxin type 2 B subunit protects mice against toxin challenge when leashed and bundled by a stable pentameric coiled-coil molecule. Vaccine. 2024 Mar 7;42(7):1757–1767</strong>). Purified Stx2eB–COMP fusion protein administered to mice conferred complete protection against a lethal dose of Stx2e. However, unfused Stx2eB conferred only 10 % protection. Furthermore, when Stx2eB was fused to a trimeric or tetrameric coiled coil, a marked reduction in protective efficacy was observed, indicating the importance of “five-to-five” fusion stoichiometry. Next, we immunized weaned piglets twice with 100 μg or 10 μg of the Stx2eB–COMP fusion protein, and then orally challenged the animals with a lethal dose of STEC. The piglets immunized with the high dose were almost completely free from clinical symptoms of ED, whereas three of the six piglets administered adjuvant-only died, and the remaining surviving piglets exhibited severe ED symptoms. Although all piglets immunized with the low dose survived, they exhibited mild to moderate ED symptoms. Our findings indicate that Stx2eB is highly protective only when bundled and molecularly stabilized by the coiled coil molecule.</div></div>\",\"PeriodicalId\":23491,\"journal\":{\"name\":\"Vaccine\",\"volume\":\"61 \",\"pages\":\"Article 127140\"},\"PeriodicalIF\":4.5000,\"publicationDate\":\"2025-07-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Vaccine\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0264410X25004372\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"IMMUNOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Vaccine","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0264410X25004372","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"IMMUNOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
猪水肿病(ED)是由产志贺毒素2e型变异(Stx2e)的大肠杆菌(STEC)感染引起的。为了开发一种新的ED疫苗,我们设计了一种融合蛋白,其中Stx2eB亚基(Stx2eB)融合到软骨寡聚基质蛋白(COMP)的五链α-螺旋螺旋结构域,基于我们最近的发现,螺旋螺旋结构强稳定Stx2的B亚基五聚体(Tamaki Y, Harakuni T, Arakawa T.)。志贺毒素2型B亚基在被稳定的五聚体螺旋螺旋分子束缚和捆绑时保护小鼠免受毒素攻击。疫苗。2024年3月7日;42(7):1757-1767。纯化的Stx2eB-COMP融合蛋白给予小鼠对致死剂量的Stx2e具有完全保护作用。然而,未融合的Stx2eB仅提供10%的保护。此外,当Stx2eB与三聚体或四聚体线圈融合时,观察到保护效果显着降低,表明“五比五”融合化学计量的重要性。接下来,我们分别用100 μg和10 μg的Stx2eB-COMP融合蛋白免疫断奶仔猪两次,然后口服致死剂量的产肠毒素大肠杆菌。注射高剂量佐剂的仔猪几乎完全没有ED的临床症状,而只注射佐剂的6头仔猪中有3头死亡,其余存活仔猪表现出严重的ED症状。虽然低剂量免疫仔猪全部存活,但均表现出轻度至中度ED症状。我们的研究结果表明,Stx2eB只有在被盘绕的线圈分子捆绑和分子稳定时才具有高度的保护作用。
B subunit of the type 2 Shiga toxin e variant (Stx2e) bundled by a five-stranded α-helical coiled coil protects piglets from porcine edema disease
Porcine edema disease (ED) is caused by infection with Shiga toxin type 2 e variant (Stx2e)-producing Escherichia coli (STEC). To develop a new ED vaccine, we engineered a fusion protein, in which the Stx2e B subunit (Stx2eB) was fused to the five-stranded α-helical coiled coil domain of cartilage oligomeric matrix protein (COMP), based on our recent finding that the coiled coil strongly stabilizes the B subunit pentamer of Stx2 (Tamaki Y, Harakuni T, Arakawa T. Shiga toxin type 2 B subunit protects mice against toxin challenge when leashed and bundled by a stable pentameric coiled-coil molecule. Vaccine. 2024 Mar 7;42(7):1757–1767). Purified Stx2eB–COMP fusion protein administered to mice conferred complete protection against a lethal dose of Stx2e. However, unfused Stx2eB conferred only 10 % protection. Furthermore, when Stx2eB was fused to a trimeric or tetrameric coiled coil, a marked reduction in protective efficacy was observed, indicating the importance of “five-to-five” fusion stoichiometry. Next, we immunized weaned piglets twice with 100 μg or 10 μg of the Stx2eB–COMP fusion protein, and then orally challenged the animals with a lethal dose of STEC. The piglets immunized with the high dose were almost completely free from clinical symptoms of ED, whereas three of the six piglets administered adjuvant-only died, and the remaining surviving piglets exhibited severe ED symptoms. Although all piglets immunized with the low dose survived, they exhibited mild to moderate ED symptoms. Our findings indicate that Stx2eB is highly protective only when bundled and molecularly stabilized by the coiled coil molecule.
期刊介绍:
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