使用阴阳性脂质体作为长dRNA的载体,在彩虹鱼细胞系中触发抗病毒反应
Shayne J Oberhoffner1, Dominique E Daniels1, Erin Cooper2
1Department of Biology, Wilfrid Laurier University, 75 University Avenue West, Waterloo, ON, N2L 3C5, Canada.
In vitro cellular & developmental biology. Animal
|January 9, 2025
概括
这项研究研究了酸性脂质体,以向鱼类细胞传递长长双链RNA (dsRNA),以提高抗病毒免疫力. 脂质体过于有毒,尽管dsrna,但无法保护病毒感染.
科学领域:
- 免疫学 免疫学 免疫学
- 分子生物学分子生物学
- 病毒学 病毒学
背景情况:
- 长双链RNA (dsRNA) 激活I型干扰素 (IFN) 和IFN刺激基因 (ISG),建立抗病毒状态.
- 这种抗病毒状态的预防性诱导可以减轻病毒感染的严重程度和死亡率.
- 需要有效的传递载体来保护dRNA在动物模型应用中的细胞外环境中的降解.
研究的目的:
- 评估商业上可用的阴阳性脂质体,用于封装并将dSRNA传递给Oncorhynchus mykiss细胞系 (RTG-2和RTgill-W1).
- 评估dsRNA载荷脂质体激活IFN通路的能力,并给予对鱼再病毒 (CSV) 感染的保护.
- 为了确定这种脂肪体dRNA输送系统在鱼细胞中的细胞毒性和有效性.
主要方法:
- 电泳运动转移试验 (EMSA) 验证dSRNA封装和稳定性.
- 用RNase III降解试验来评估dRNA的保护.
- 细胞活力测试用于评估脂质体-dsRNA细胞毒性.
- Mx1基因表达分析以测量IFN通路激活.
- 病毒挑战测试与鱼重病毒 (CSV) 测试,以确定抗病毒保护.
主要成果:
- 脂质体在55和800微克/毫升时有效封装dRNA,在4°C和-20°C显示稳定性.
- 封装的dsRNA受到了RNase III降解的保护.
- 装有dsRNA的脂质体在24小时暴露后表现出高细胞毒性.
- 2小时的暴露降低了细胞毒性和增强了Mx1表达,但不足以保护病毒.
- 未封装的dsRNA显著降低了CSV标位和突触形成.
结论:
- 测试的脂质体配方对于在鱼细胞中有效的抗病毒应用过于细胞毒.
- 尽管传递载体存在挑战,但dRNA仍然是鱼类中强大的免疫调节器.
- 需要进一步的研究来开发更少的毒性和更有效的dsrna输送系统用于水产养殖.
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