C19 SARS-CoV-2-specific T cells in the infected nasel epithelium

总结 对SARS-CoV-2感染的获得性抗原特异性免疫反应可能是 确定COVID-19的结果以及疫苗接种的有效性和持久性。cd 4 T细胞 需要帮助才能产生持久的、强烈的抗体反应。主要候选疫苗包括 - 也可以诱导CD 8 T细胞的一些形式，如病毒载体和核酸，以及 其它形式如佐剂化蛋白质不能。抗病毒药物的有力证据 病毒特异性CD 8 T细胞的作用可能会使平衡向有利于CD 8诱导疫苗的方向倾斜 平台本探索性R21对T细胞对SARS-CoV-2应答的目的1和目的2 分别涉及CD 8和CD 4 T细胞应答。上下呼吸道 上皮是SARS-CoV-2的主要靶组织。鼻上皮特别是 富含病毒进入所需的宿主分子ACE 2和TMPRSS 2的表达。我们利用 合作研究者Abuzeid在获取鼻细胞和扩大原发性鼻肿瘤方面的经验 上皮细胞（HNEpC）和合作者Greninger在BSL 3病毒学和 定量RNA测量以创建研究T细胞感染的上皮细胞的系统 体外相互作用我们利用我们的大型COVID-19生物库、实验室内经验和其他NIAID 资源，以产生功能上无限量的高亲和力SARS-CoV-2特异性CD 8 和具有精确已知的HLA限制和确定的肽靶的CD 4 T细胞。感染 HNEpC将被仔细表征和优化。然后我们将测量T细胞效应子 功能包括杀死感染的细胞、减少病毒子代输出和细胞因子 release.在后续研究中，我们希望利用SARS-CoV-2基因系统， 在候选免疫逃避基因中定义突变体，并将我们的研究扩展到下呼吸道 束细胞和类器官。总的来说，我们努力模拟免疫系统之间的重要相互作用。 细胞和SARS-CoV-2感染的呼吸道。 相关性 疫苗可能需要引发SARS-CoV-2特异性T细胞，这些T细胞可以快速识别SARS-CoV-2。 2-特异性感染上呼吸道上皮细胞，并关闭其生产 子代病毒，以限制患者体内感染和症状的进展，以及 减少可能传染他人的传染性病毒的散播。在R21补助金中 应用，我们将使用既定的工作方法来隔离大量的纯SARS- 来自康复者的CoV-2特异性T细胞并研究其发挥抗病毒作用的能力 使用细胞培养物的细胞，这些细胞排列在鼻腔中，通常在COVID期间被感染， 19.

Summary The acquired, antigen-specific immune response to SARS-CoV-2-infection is likely central to determining the outcome of COVID-19 and the efficacy and durability of vaccination. CD4 T cell help is required for lasting, avid antibody responses. The leading vaccine candidates include some formats that can also induce CD8 T cells, such as viral vectors and nucleic acids, and other formats such as adjuvanted protein that cannot. Compelling evidence for an antiviral effects of virus-specific CD8 T cells could tilt the balance in favor of CD8-eliciting vaccine platforms. Aim 1 and Aim 2 of this exploratory R21 on the T cell response to SARS-CoV-2 concern CD8- and CD4 T cell responses, respectively. Upper and lower airway respiratory epithelium are the primary target tissues for SARS-CoV-2. The nasal epithelium is particularly rich in expression of ACE2 and TMPRSS2, host molecules required for viral entry. We leverage Co-Investigator Abuzeid’s experience in harvesting nasal cells and expanding primary nasal epithelial cells (HNEpC) and Collaborator Greninger’s expertise in BSL3 virology and quantitative RNA measurement to create systems to study T cell-infected epithelial cell interactions in vitro. We use our large COVID-19 biobank, in-lab experience, and other NIAID resources to generate functionally unlimited amounts of highly avid SARS-CoV-2-specific CD8 and CD4 T cells with precisely known HLA restriction and defined peptide targets. Infection of HNEpC will be carefully characterized and optimized. We will then measure T cell effector functions including the killing of infected cells, reduction of viral progeny output, and cytokine release. In follow-up studies, we hope to leverage SARS-CoV-2 genetic systems to make defined mutants in candidate immune evasion genes and extend our studies to lower respiratory tract cells and organoids. Overall, we strive to model important interactions between immune cells and the SARS-CoV-2 infected respiratory tract. Relevance Vaccines may need to elicit SARS-CoV-2 specific T cells that can quickly recognize SARS-CoV- 2-specific infected upper respiratory tract epithelial cells and shut down their production of progeny virus, to both limit the progression of infection and symptoms within patients, and reduce the shedding of infectious virus that may infect other persons. In this R21 grant application, we will use established, working methods to isolate large quantities of pure SARS- CoV-2-specific T cells from recovered persons and study their ability to exert anti-viral effects using cell cultures of the cells that line the nasal cavity that are normally infected during COVID- 19.