单壁碳纳米管载嗜水气单胞菌外膜蛋白亚单位疫苗的研制及免疫效果评价
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S942.5

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国家自然科学基金(31670129)


Development of single-walled carbon nanotubes carrying Aeromonas hydrophila outer membrane protein subunit vaccine and evaluation of immune effect
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    摘要:

    为了筛选细菌的疫苗候选蛋白,本实验根据课题组前期的研究,挑选了2个嗜水气单胞菌外膜蛋白(A0KLQ6和A0KH89),以A0KHR9(OmpAII)为对照,通过比较外膜蛋白与碳纳米管载外膜蛋白通过腹腔注射与浸泡免疫模式,比较和评价斑马鱼的免疫反应与免疫保护效果。结果显示,腹腔注射组和浸泡组与普通蛋白组对比,碳纳米管载蛋白能够显著提高斑马鱼免疫相关基因表达量。腹腔注射组在5 μg剂量下免疫A0KHR9、A0KLQ6和A0KH89蛋白分别获得61.60%、65.39%和84.96%相对免疫保护率(relative immune protection rate,RPS),在5 μg剂量下免疫碳纳米管载蛋白SWCNTs-A0KHR9、SWCNTs-A0KLQ6和SWCNTs-A0KH89分别获得68.10%、77.50%和90.43%的RPSs。浸泡组在40 mg/L剂量下免疫A0KHR9、A0KLQ6和A0KH89蛋白分别获得30.80%、25.85%和37.80%的RPSs,在40 mg/L最高剂量下免疫碳纳米管载蛋白疫苗SWCNTs-A0KHR9、SWCNTs-A0KLQ6和SWCNTs-A0KH89分别获得63.60%、73.74%和68.49%的RPSs。研究表明,铁离子限制下嗜水气单胞菌外膜蛋白A0KLQ6和A0KH89等能够作为抗嗜水气单胞菌的亚单位疫苗候选蛋白,而单壁碳纳米管载蛋白通过注射免疫与浸泡免疫均能显著提升免疫疗效,是一种疫苗的适宜纳米载体。以上研究为寻找高效的嗜水气单胞菌亚单位疫苗候选蛋白及佐剂提供了理论依据。

    Abstract:

    To investigate the immune response of zebrafish to iron-related proteins and SWCNT-encapsulated proteins, we overexpressed and purified two iron-related recombinant proteins (A0KLQ6 and A0KH89) from Aeromonas hydrophila. We then vaccinated Danio rerio with these proteins and their SWCNT-encapsulated counterparts via both intraperitoneal injection and immersion immunization, and found they evoked a strong immune response in D. rerio. When challenged with virulent A. hydrophila, D. rerio administered 5 μg intraperitoneal injections of A0KHR9, A0KLQ6, A0KH89, SWCNTs-A0KHR9, SWCNTs-A0KLQ6 and SWCNTs-A0KH89 obtained high relative percent survivals (RPSs) (61.60%, 65.39%, 84.96%, 68.10%, 77.50% and 90.43%, respectively). Meanwhile, the D. rerio administered 40 mg/L immersion immunizations of aboved samples had 30.80%, 25.85%, 37.80%, 63.60%, 73.74% and 68.49% RPSs, respectively. These results indicate that the outer membrane proteins A0KLQ6 and A0KH89 can be used as potenial subunit vaccine candidates against A. hydrophila infection, and the SWCNTs-encapsulated counterparts can significantly improve the immune effect in both immunization ways. Our studies may provide a theoretical basis for the development of vaccines and adjuvants candidates

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姜倩雯,张丽珊,李小艳,武瑶,赵怡扬,徐慧铭,林向民.单壁碳纳米管载嗜水气单胞菌外膜蛋白亚单位疫苗的研制及免疫效果评价[J].水产学报,2020,44(9):1424~1434

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  • 收稿日期:2019-04-20
  • 最后修改日期:2019-06-27
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  • 在线发布日期: 2020-09-27
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