mRNA induced expression of human angiotensin-converting enzyme 2 in mice for the study of the adaptive immune response to severe acute respiratory syndrome coronavirus 2.
mRNA induced expression of human angiotensin-converting enzyme 2 in mice for the study of the adaptive immune response to severe acute respiratory syndrome coronavirus 2.
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DOI：
10.1371/journal.ppat.1009163
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发表时间：
2020-12
期刊：
影响因子：
6.7
通讯作者：
Pinto AK
中科院分区：
文献类型：
--
作者：
Hassert M;Geerling E;Stone ET;Steffen TL;Feldman MS;Dickson AL;Class J;Richner JM;Brien JD;Pinto AK
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）是一种新型的人类冠状病毒，它已引起了一场大流行。快速评估SARS-CoV-2疫苗和抗病毒药物的关键是开发易于处理的动物模型，以了解对病毒的适应性免疫反应。为此，由于SARS-CoV-2进入所需的受体血管紧张素转换酶2（ACE 2）的显著差异，普通实验室小鼠品系的使用受到阻碍。在目前的研究中，我们设计并利用基于mRNA的转染系统来诱导hACE 2受体的表达，以使SARS-CoV-2进入其他非许可细胞。通过在体内环境中使用该表达系统，我们能够询问1型干扰素受体缺陷小鼠对SARS-CoV-2的适应性免疫应答。在这样做的过程中，我们发现，当hACE 2在感染过程中表达时，T细胞对SARS-CoV-2的反应增强。此外，我们证明了这些反应被保存在记忆中，并在二次感染时得到加强。重要的是，使用该系统，我们功能性地鉴定了H2 b限制性小鼠中SARS-CoV-2感染期间靶向的CD 4+和CD 8+结构肽表位，并证实了它们在已建立的SARS-CoV-2发病机制模型中的存在。我们证明，与在人类中观察到的相同，小鼠中的抗原特异性CD 8 + T细胞主要靶向刺突和膜蛋白的肽，而抗原特异性CD 4 + T细胞靶向核衣壳、膜和刺突蛋白的肽。由于小鼠免疫应答的焦点与人类免疫应答的焦点高度相似，因此本研究中提供的功能性鼠SARS-CoV-2特异性T细胞表位的鉴定对于评价疫苗在鼠SARS-CoV-2感染模型中的效力至关重要。易处理的小动物模型的开发对于了解对新型人类冠状病毒严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的免疫应答以及评估针对该病毒的疫苗至关重要。然而，由于缺乏人血管紧张素转换酶2（hACE 2）的表达，小鼠感染模型的开发受到阻碍，hACE 2是SARS-CoV-2进入所需的受体。在这项研究中，我们将hACE 2基因克隆到mRNA表达载体中，并证明转染该mRNA允许SARS-CoV-2进入和复制。我们利用这种通过体内mRNA转染在小鼠中表达hACE 2的新方法来表征对SARS-CoV-2的适应性免疫应答。这种独特而易处理的模型允许首次表征鼠SARS-CoV-2特异性T细胞应答。这些信息对于确定抗病毒保护的相关性和评估疫苗至关重要。
The novel human coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a pandemic. Critical to the rapid evaluation of vaccines and antivirals against SARS-CoV-2 is the development of tractable animal models to understand the adaptive immune response to the virus. To this end, the use of common laboratory strains of mice is hindered by significant divergence of the angiotensin-converting enzyme 2 (ACE2), which is the receptor required for entry of SARS-CoV-2. In the current study, we designed and utilized an mRNA-based transfection system to induce expression of the hACE2 receptor in order to confer entry of SARS-CoV-2 in otherwise non-permissive cells. By employing this expression system in an in vivo setting, we were able to interrogate the adaptive immune response to SARS-CoV-2 in type 1 interferon receptor deficient mice. In doing so, we showed that the T cell response to SARS-CoV-2 is enhanced when hACE2 is expressed during infection. Moreover, we demonstrated that these responses are preserved in memory and are boosted upon secondary infection. Importantly, using this system, we functionally identified the CD4+ and CD8+ structural peptide epitopes targeted during SARS-CoV-2 infection in H2b restricted mice and confirmed their existence in an established model of SARS-CoV-2 pathogenesis. We demonstrated that, identical to what has been seen in humans, the antigen-specific CD8+ T cells in mice primarily target peptides of the spike and membrane proteins, while the antigen-specific CD4+ T cells target peptides of the nucleocapsid, membrane, and spike proteins. As the focus of the immune response in mice is highly similar to that of the humans, the identification of functional murine SARS-CoV-2-specific T cell epitopes provided in this study will be critical for evaluation of vaccine efficacy in murine models of SARS-CoV-2 infection. The development of tractable small animal models is critical to gain an understanding of the immune response to the novel human coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and for the evaluation of vaccines against the virus. However, the development of murine models of infection has been hindered due to the lack of expression of the human angiotensin-converting enzyme 2 (hACE2), which is the receptor required for entry of SARS-CoV-2. In this study, we cloned the hACE2 gene into an mRNA expression vector and demonstrated that transfection with this mRNA allowed for SARS-CoV-2 entry and replication. We utilized this novel method of hACE2 expression in mice by in vivo mRNA transfection to characterize the adaptive immune response to SARS-CoV-2. This unique and tractable model allowed for the first ever characterization of the murine SARS-CoV-2 specific T cell response. This information will be critical to determining the correlates of protection against the virus and for the evaluation of vaccines.
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影响因子：
64.8
作者：
Li W;Moore MJ;Vasilieva N;Sui J;Wong SK;Berne MA;Somasundaran M;Sullivan JL;Luzuriaga K;Greenough TC;Choe H;Farzan M
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影响因子：
5.4
作者：
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作者：
Kuba K;Imai Y;Rao S;Gao H;Guo F;Guan B;Huan Y;Yang P;Zhang Y;Deng W;Bao L;Zhang B;Liu G;Wang Z;Chappell M;Liu Y;Zheng D;Leibbrandt A;Wada T;Slutsky AS;Liu D;Qin C;Jiang C;Penninger JM
通讯作者：
Penninger JM
影响因子：
82.9
作者：
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通讯作者：
Krammer F