Virus-like particles displaying conserved toxin epitopes stimulate broadly reactive, polyspecific, murine antibody responses capable of snake venom recognition
Virus-like particles displaying conserved toxin epitopes stimulate broadly reactive, polyspecific, murine antibody responses capable of snake venom recognition
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DOI：
10.21203/rs.3.rs-1044937/v1
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发表时间：
2021-11
期刊：
影响因子：
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通讯作者：
S. Menzies;Charlotte A. Dawson;E. Crittenden;Rebecca J. Edge;Steven R. Hall;Jaffer Alsolaiss;M. Wilkinson;N. Casewell;R. Harrison;S. Ainsworth
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文献类型：
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作者：
S. Menzies;Charlotte A. Dawson;E. Crittenden;Rebecca J. Edge;Steven R. Hall;Jaffer Alsolaiss;M. Wilkinson;N. Casewell;R. Harrison;S. Ainsworth
抗蛇毒血清是目前治疗毒蛇咬伤的首选药物。然而，只有低比例的抗蛇毒免疫球蛋白对毒液毒素具有特异性，导致剂量功效和效力差。我们试图研究在病毒样颗粒上展示的线性毒液表位是否可以刺激能够识别来自不同医学重要物种的毒液毒素的稳健和集中的抗体应答。生物信息学设计的表位，对应于预测的保守区域的组I磷脂酶A2和三指毒素，被工程化展示在表面上的肝炎B核心抗原病毒样颗粒，并用于免疫雌性CD 1小鼠超过14周。所有毒液表位病毒样颗粒的抗体应答可通过ELISA在免疫期结束时检测到，尽管针对不同表位免疫原的总抗体和表位特异性抗体滴度是可变的。使用汇集血清的免疫印迹证明了在六个眼镜蛇毒液，代表三大洲和四个属的不同面板中的各种毒液成分的识别。最后，汇集终端血清进行了比较，通过定量免疫印迹常规抗蛇毒血清，并证明上级识别低分子量眼镜蛇毒液毒素。这项研究证明了原理证明，病毒样颗粒工程化展示保守的毒素线性表位可以引发特异性抗体反应，小鼠能够识别地理上广泛的眼镜蛇毒液。
Antivenom is currently the first-choice treatment for snakebite envenoming. However, only a low proportion of antivenom immunoglobulins are specific to venom toxins, resulting in poor dose efficacy and potency. We sought to investigate whether linear venom epitopes displayed on virus like particles can stimulate a robust and focused antibody response capable of recognising venom toxins from diverse medically important species. Bioinformatically-designed epitopes, corresponding to predicted conserved regions of group I phospholipase A2 and three finger toxins, were engineered for display on the surface of hepatitis B core antigen virus like particles and used to immunise female CD1 mice over a 14-weeks. Antibody responses to all venom epitope virus like particles were detectable by ELISA by the end of the immunisation period, although total antibody and epitope specific antibody titres were variable against the different epitope immunogens. Immunoblots using pooled sera demonstrated recognition of various venom components in a diverse panel of six elapid venoms, representing three continents and four genera. Finally, pooled terminal sera was compared to conventional antivenom via quantitative immunoblot, and demonstrated superior recognition of lower-molecular weight elapid venom toxins. This study demonstrates proof-of-principle that virus like particles engineered to display conserved toxin linear epitopes can elicit specific antibody responses in mice which are able to recognise a geographically broad range of elapid venoms.