由SARS-CoV-2引起的COVID-19在全球产生了重大影响。虽然疫苗和治疗减少了严重病例和死亡,长期影响尚未得到很好的理解。目前用于研究的模型,如非人灵长类动物和转基因小鼠,价格昂贵,需要稀缺的生物安全3级(BSL-3)实验室,从而限制了其实用性。然而,小鼠肝炎病毒1(MHV-1)小鼠模型提供了一个有希望的替代方案。该替代模型可以在更广泛可用的生物安全级别2(BSL-2)实验室中进行研究。此外,老鼠经济实惠,易于处理,利用MHV-1作为SARS-CoV-2的替代品消除了对昂贵的转基因小鼠的需要。重要的是,MHV-1模型成功地概括了COVID-19相关的临床症状,减肥,急性期多器官病理变化和衰竭,不可逆的神经系统并发症,和其他感染后的长期器官功能障碍,这与COVID-19后可用的人类数据相似。为了帮助研究人员建立和使用MHV-1小鼠模型,该协议提供了全面的指导,包括动物准备程序,诱导病毒感染,临床观察,病理变化,和组织分析用于机械研究,从而对疾病机制和进展产生有价值的见解。通过采用MHV-1模型和提供的协议,研究人员可以有效地规避资金限制和BSL-3实验室的有限可用性,从而促进更容易获得和成本效益的方法来调查SARS-CoV-2病理生理学的潜在机制和探索潜在的治疗干预措施.©2023威利期刊有限责任公司。基本方案:在A/J小鼠中诱导小鼠肝炎病毒1(MHV-1)感染支持方案1:组织学评估支持方案2:肝酶测量支持方案3:水通道蛋白表达的蛋白质印迹分析支持方案4:mRNA测量支持方案5:免疫组织化学/免疫荧光支持方案6:组织水测量。
COVID-19, caused by SARS-CoV-2, has had a significant global impact. While vaccines and treatments have reduced severe cases and deaths, the long-term effects are not yet well understood. Current models used for research, such as non-human primates and transgenic mice, are expensive and require scarce Biosafety Level-3 (BSL-3) laboratories, thereby limiting their practicality. However, the mouse hepatitis virus 1 (MHV-1) mouse model offers a promising alternative. This surrogate model can be investigated in more widely available Biosafety Level-2 (BSL-2) laboratories. Furthermore, mice are affordable and easy to handle, and utilizing MHV-1 as a surrogate for SARS-CoV-2 eliminates the need for costly transgenic mice. Importantly, the MHV-1 model successfully recapitulates COVID-19-related clinical symptoms, weight loss, multiorgan pathological changes and failure in acute stages, irreversible neurological complications, and other long-term organ dysfunction post-infection, which are similar to available human data post-COVID-19. To assist researchers in establishing and using the MHV-1 mouse model, this protocol offers comprehensive guidance encompassing procedures for animal preparation, induction of viral infection, clinical observation, pathological changes, and tissue analysis for mechanistic studies, thereby yielding valuable insights into disease mechanisms and progression. By adopting the MHV-1 model and the provided protocols, researchers can effectively circumvent financial constraints and the limited availability of BSL-3 laboratories, thus facilitating a more accessible and cost-effective approach to investigating the underlying mechanisms of SARS-CoV-2 pathophysiology and exploring potential therapeutic interventions. © 2023 Wiley Periodicals LLC. Basic Protocol: Induction of mouse hepatitis virus 1 (MHV-1) infection in A/J mice Support Protocol 1: Histological evaluation Support Protocol 2: Liver enzyme measurement Support Protocol 3: Western blot analysis of aquaporin expression Support Protocol 4: mRNA measurement Support Protocol 5: Immunohistochemistry/immunofluorescence Support Protocol 6: Tissue water measurement.