SARS-CoV-2-reactive tissue-resident memory T cells in healthy and cancer subjects

项目总结 这份名为“SARS-CoV-2-Reactive”的多PI提案 主题“是根据‘RFA-CA-20-039’编写的- 健康人和癌症患者的组织驻留记忆T细胞 SARS-CoV-2血清学研究项目。 最近的研究表明，对SARS-CoV-2感染的抗体反应随着时间的推移迅速下降，这意味着 缺乏持久的保护性体液(B细胞)免疫。细胞免疫是否也是如此(例如，T 细胞)对此知之甚少。众所周知，CD8TRM细胞是病毒感染的第一道防线 在粘膜/屏障部位。众所周知，它们还可以保护宿主免受同源或异源的再次感染。 我们小组是第一个证明TRM细胞在驱动有效的抗肿瘤免疫反应中起关键作用的人 而TRM细胞是抗PD1治疗的主要细胞靶点。这些主要发现是 由于Vijayanand博士、Ay博士和Ottensmeier博士(多PI)之间的持续合作，这是可能的。 这个团队汇集了T细胞免疫学、单细胞基因组学、生物信息学和癌症方面的经验 免疫学。我们的多PI团队最近完成了第一个也是最大的单细胞RNA-SEQ和TCR-SEQ 新冠肺炎患者SARS-CoV-2反应性CD8T细胞和CD4T细胞(约30万单细胞)分析这里, 为了了解TRM对SARS-CoV-2的反应，我们将利用癌症(n=100)和非癌症的队列 (n=100)患者，将提供多余的呼吸道(鼻、口咽、喉)、肺和肿瘤组织标本 在常规手术中获得的。在目标1中，我们将定义SARS-CoV-2反应性TRM细胞的特性 有或无SARS-CoV-2感染史的癌症和非癌症患者。我们将联合表演 呼吸道(鼻腔、口咽)、肺和肿瘤中CD8 TRM细胞的单细胞RNA-SEQ和TCR-SEQ分析 组织。同时，通过用SARS-CoV-2肽库刺激PBMC，我们将确定转录转录 SARS-CoV-2反应性T细胞的TCR序列。我们将利用此TCR序列信息来定义 粘膜和肿瘤组织中SARS-CoV-2反应性TRM细胞的数量和特性。最新研究在非- 接触过病毒的人(新冠肺炎大流行前)表明已存在循环中的CD8T细胞，与人类 冠状病毒交叉反应。在这里，我们将衡量以前存在的SARS-CoV-2交叉病毒的数量和质量。 无SARS-CoV-2感染的临床或血清学证据的受试者的反应性TRM反应。在……里面 目的2，我们将评估SARS-CoV-2感染对抗肿瘤和其他抗病毒TRM反应的影响。我们 将以(I)常见呼吸道RNA病毒为靶点的多肽库刺激匹配的PBMC(如上) (流感(流感)，RSV)，(Ii)持久性DNA病毒(CMV，EBV)，和(Iii)肿瘤驱动病毒(HPV)来定义 病毒特异性和肿瘤(HPV)特异性CD8T细胞的TCR序列；我们将利用TCR 确定粘膜和肿瘤中其他病毒/肿瘤反应性TRM细胞的频率和特性的信息- 组织细胞。

PROJECT SUMMARY This multi-PI proposal titled “SARS-CoV-2-reactive subjects” is written in response to ‘RFA-CA-20-039’ - tissue-resident memory T cells in healthy and cancer Research projects in SARS-CoV-2 Serological Sciences. Recent studies have shown that antibody responses to SARS-CoV-2 infection decline rapidly over time, implying a lack of durable protective humoral (B cell) immunity. Whether this is also true for cellular immunity (e.g., T cells) is poorly understood. It is well established that CD8+ TRM cells are the first line of defense in viral infections at mucosal/barrier sites. They are also known to protect hosts against homologous or heterologous re-infections. Our group was the first to show that TRM cells are pivotal players in driving effective anti-tumor immune responses in lung cancer, and that TRM cells are the primary cellular targets of anti-PD1 therapies. These key findings were possible because of the ongoing collaboration between Dr. Vijayanand, Dr. Ay, and Dr. Ottensmeier (Multi-PI). This team brings together experience in T cell immunology, single-cell genomics, bioinformatics, and cancer immunology. Our Multi-PI team has recently performed the first and largest single-cell RNA-seq and TCR-seq analysis of SARS-CoV-2-reactive CD8+ and CD4+ T cells (~300,000 single-cells) from COVID-19 patients. Here, to understand TRM responses to SARS-CoV-2, we will capitalize on a cohort of cancer (n=100) and non-cancer (n=100) patients, who will provide excess airway (nasal, oropharynx, larynx), lung and tumor tissue specimens obtained during routine surgery. In AIM 1, we will define the properties of SARS-CoV-2 reactive TRM cells from cancer and non-cancer patients with or without previous SARS-CoV-2 infection. We will perform combined single-cell RNA-seq and TCR-seq analysis of CD8+ TRM cells in the airways (nasal, oropharynx), lung, and tumor tissue. In parallel, by stimulating PBMCs with SARS-CoV-2 peptide pool, we will determine the transcriptomic and TCR sequence of SARS-CoV-2 reactive T cells. We will utilize this TCR sequence information to define the numbers and properties of SARS-CoV-2 reactive-TRM cells in mucosal and tumor tissues. Recent studies in non- exposed individuals (pre-COVID-19 pandemic) indicate pre-existing, circulating CD8+ T cells, with human coronavirus cross-reactivity. Here, we will measure the quantity and quality of pre-existing SARS-CoV-2 cross- reactive TRM responses in subjects without clinical or serological evidence of previous SARS-CoV-2 infection. In AIM 2, we will assess the impact of SARS-CoV-2 infection on anti-tumor and other anti-viral TRM responses. We will stimulate matched PBMCs (as above) with peptide pools targeting (i) common respiratory RNA viruses (influenza (FLU), RSV), (ii) persistent DNA viruses (CMV, EBV), and (iii) a tumor-driving virus (HPV) to define the TCR sequence of the respective virus-specific and tumor(HPV)-specific CD8+ T cells; we will utilize the TCR information to determine frequency and properties of other virus/tumor-reactive TRM cells in mucosal and tumor- tissue cells.