自强化RNA的阿尔法病毒骨干增强了对SARS-CoV-2抗原的蛋白质表达和免疫性
Irafasha C Casmil1, Nuthan V Bathula1, Cynthia Huang1
1Michael Smith Laboratories, University of British Columbia, Vancouver V6T1Z4, BC, Canada; School of Biomedical Engineering, University of British Columbia, Vancouver V6T1Z4, BC, Canada.
概括
这项研究引入了来自各种阿尔法病毒的新型自我放大RNA (saRNA) 载体. 这些工程化saRNA载体显示可调节的蛋白质表达和可变的免疫性,为先进的疫苗和疗法铺平了道路.
科学领域:
- 分子生物学分子生物学
- 病毒学 病毒学
- 疫苗学 疫苗学 疫苗学
背景情况:
- 自放大RNA (saRNA) 是一种用于增强基因表达的先进RNA技术.
- 目前的saRNA疫苗开发受限于一系列狭窄的alphavirus骨干,只有委内瑞拉马类脑炎病毒基载体在临床上使用.
- 与信使RNA (mRNA) 相比,saRNA在疫苗中的剂量节约潜力已在临床上得到证实.
研究的目的:
- 设计和表征新的,基因型上不同的α病毒saRNA载体.
- 评估这些saRNA载体的表现,包括蛋白质表达和免疫性,在体外和体内.
- 通过检查它们在人类皮肤实验中的表现来评估saRNA载体的翻译潜力.
主要方法:
- 设计和建造12个不同的α病毒saRNA载体.
- 在哺乳动物细胞系中的体外表征.
- 在体内评估转基因表达和小鼠的药理动力学.
- 在不同小鼠菌株中评估幽默和细胞免疫性.
- 在人体皮肤实验中对saRNA载体性能进行了ex vivo分析.
主要成果:
- 在12个设计的saRNA载体中,有5个在小鼠中表现出显著的露西法酶表达,表达大小和持续时间可调.
- 发现saRNA载体的α病毒基因型显著影响疫苗免疫性,影响小鼠的幽默和细胞反应.
- 当在人体皮肤中测试saRNA载体时,观察到高的转基因表达.
- 在不同的小鼠菌株中,saRNA生物发光和免疫性具有很强的相关性,但与人类的扩散物或细胞系数据的相关性很小.
结论:
- 基因多样化的α病毒可以适应为有效的saRNA载体,用于疫苗和治疗应用.
- 选择阿尔法病毒基因型对于调节saRNA载体性能和免疫性至关重要.
- 虽然小鼠模型为免疫原性提供了相关数据,但人类皮肤实验对评估与人类应用相关的转基因表达有价值.
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