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

摘要 针对SARS-CoV-2感染的获得性抗原特异性免疫反应可能是 决定新冠肺炎的结果和疫苗接种的有效性和持久性。CD4T细胞 持久的、狂热的抗体反应需要帮助。领先的候选疫苗包括 一些也可以诱导CD8T细胞的形式，如病毒载体和核酸，以及 其他形式，如佐剂蛋白质，不能。抗病毒药物的令人信服的证据 病毒特异性CD8T细胞的作用可能会使天平向CD8疫苗倾斜 站台。本探索性R21的目的1和目的2关于T细胞对SARS-CoV-2的应答 分别关注CD8和CD4T细胞反应。上、下呼吸道 上皮细胞是SARS-CoV-2的主要靶组织。鼻腔上皮特别是 丰富表达ACE2和TMPRSS2，这是病毒进入所需的宿主分子。我们利用 联合调查员阿布泽德在获取鼻细胞和扩大原代鼻腔方面的经验 上皮细胞(HNEpC)及其合作者Griinger在BSL3病毒学和 定量RNA测量建立研究T细胞感染的上皮细胞的系统 体外相互作用。我们使用我们的大型新冠肺炎生物库、实验室内经验和其他NIAID 产生高度狂热的SARS-CoV-2特异性CD8的功能不限数量的资源 和CD4T细胞，具有确切的已知的人类白细胞抗原限制和确定的多肽靶标。感染性疾病 HNEpC将被仔细描述和优化。然后我们将测量T细胞效应器 功能包括杀死受感染的细胞，减少病毒后代的产量，以及细胞因子 放手。在后续研究中，我们希望利用SARS-CoV-2基因系统使 确定候选免疫逃避基因的突变，并将我们的研究扩展到下呼吸道 束细胞和细胞器。总体而言，我们努力对免疫之间的重要相互作用进行建模 细胞和SARS-CoV-2感染的呼吸道。 相关性 疫苗可能需要诱导出能够快速识别SARS-CoV-2的特异性T细胞-- 2-特异性感染上呼吸道上皮细胞，并停止其产生 子代病毒，既可以限制感染的进展，也可以限制患者的症状，以及 减少可能感染他人的传染性病毒的脱落。在这笔R21拨款中 应用，我们将使用既定的工作方法来分离大量的纯SARS- 康复者CoV-2特异性T细胞及其抗病毒作用的研究 使用通常在冠状病毒感染期间感染的鼻腔细胞的细胞培养- 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.