Zheng Jian, Channappanavar Rudra, Perlman Stanley
Department of Microbiology and Immunology, School of Medicine, University of Louisville, Louisville, KY 40202, USA; Center for Predictive Medicine, School of Medicine, University of Louisville, Louisville, KY 40202, USA; Departments of Microbiology and Immunology, University of Iowa, Iowa City, IA 52242, USA.
Department of Veterinary Pathobiology, Oklahoma State University, Stillwater, OK, USA.
STAR Protoc. 2023 Feb 27;4(2):102171. doi: 10.1016/j.xpro.2023.102171.
Here, we detail the immunization of mice with a sublethal dose of MERS-CoV or two doses of replication-incompetent alphavirus replicon particles expressing MERS-CoV spike protein. We then describe steps to determine the outcome of immunization by challenging immunized mice with a lethal dose of MERS-CoV, as well as by detecting virus-specific neutralizing antibody and virus-specific T cell response via neutralization assay and flow cytometry, respectively. This protocol can be used to evaluate other CoV infections or vaccine-induced immune responses. For complete details on the use and execution of this protocol, please refer to Zheng et al. (2021)..
在此，我们详细介绍了用亚致死剂量的中东呼吸综合征冠状病毒（MERS-CoV）或两剂表达MERS-CoV刺突蛋白的无复制能力的甲病毒复制子颗粒对小鼠进行免疫接种的过程。然后，我们描述了通过用致死剂量的MERS-CoV攻击免疫小鼠，以及分别通过中和试验和流式细胞术检测病毒特异性中和抗体和病毒特异性T细胞反应来确定免疫结果的步骤。该方案可用于评估其他冠状病毒感染或疫苗诱导的免疫反应。有关该方案使用和执行的完整详细信息，请参考郑等人（2021年）的研究。
