Human Coronary Plaque T Cells Are Clonal and Cross-React to Virus and Self.
Human Coronary Plaque T Cells Are Clonal and Cross-React to Virus and Self.
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人冠状动脉斑块T细胞是克隆性的，并与病毒和自身交叉反应。
DOI：
10.1161/circresaha.121.320090
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发表时间：
2022-05-13
影响因子：
20.1
通讯作者：
Nguyen, Patricia K.
中科院分区：
文献类型：
--
作者：
Chowdhury, Roshni Roy;D'Addabbo, Jessica;Huang, Xianxi;Veizades, Stefan;Sasagawa, Koki;Louis, David M.;Cheng, Paul;Sokol, Jan;Jensen, Annie;Tso, Alexandria;Shankar, Vishnu;Wendel, Ben Shogo;Bakerman, Isaac;Liang, Grace;Koyano, Tiffany;Fong, Robyn;Nau, Allison N.;Ahmad, Herra;Gopakumar, Jayakrishnan;Wirka, Robert;Lee, Andrew S.;Boyd, Jack;Woo, Y. Joseph;Quertermous, Thomas;Gulati, Gunsagar Singh;Jaiswal, Siddhartha;Chien, Yueh-Hsiu;Chan, Charles Kwok Fai;Davis, Mark M.;Nguyen, Patricia K.
关键词：
冠状动脉疾病曾经被认为主要是一种脂质沉积性疾病，是一种无法治愈、危及生命的疾病，现在其特征还在于慢性炎症，其特征是动脉粥样硬化斑块的形成，其中含有处于各种激活和分化状态的免疫细胞。了解这些免疫细胞如何促进疾病进展可能有助于开发新的治疗策略。我们使用单细胞技术和体外测定来研究不同成熟阶段的人冠状动脉粥样硬化斑块的免疫微环境。除了巨噬细胞之外，我们还在冠状动脉斑块中发现了高比例的 αβ T 细胞。大多数这些 T 细胞缺乏 CCR7 和 L-选择素的高表达，表明它们主要是“经历过抗原”的记忆细胞。值得注意的是，这些细胞中近三分之一表达 HLA-DRA 表面标记，表明通过其 T 细胞抗原受体 (TCR) 进行激活。与此一致的是，TCR 库分析证实了活化的 αβ T 细胞 (CD4 << CD8) 的存在，表现出特定 TCR 的克隆扩增。有趣的是，我们发现这些斑块T细胞具有针对流感、冠状病毒和其他病毒表位的特异性TCR，这些表位与平滑肌细胞和内皮细胞上发现的蛋白质具有序列同源性，这表明在没有主动感染的情况下潜在的自身免疫介导的T细胞激活。为了更好地了解这些激活的斑块 T 细胞的潜在功能，我们在单细胞水平上研究了它们的转录组。在 Seurat 算法鉴定出的激活标记 HLA-DRA 表达最高的 3 个 T 细胞表型簇中，两个簇表达 CD8 细胞的促炎和溶细胞特征，而另一个则表达双调蛋白，促进平滑肌细胞增殖和纤维化，从而有助于斑块进展。总而言之，这些发现表明，斑块 T 细胞可能通过抗原结合进行克隆扩增，可能对自身表位发生反应，并且可能与斑块微环境中的平滑肌细胞和巨噬细胞相互作用。尽管最近使用单细胞 RNA 测序的研究已经表征了动脉粥样硬化斑块内的 T 细胞，但这些分析仅限于小鼠主动脉病变和晚期人类颈动脉斑块。虽然动脉粥样硬化患者血液中的 T 细胞克隆已被确定，但斑块 T 细胞库仍有待分析。在这里，我们提供了人类冠状动脉斑块 T 细胞库及其跨疾病谱的转录组的第一个综合图谱。我们证明 T 细胞效应记忆簇占主导地位，表达激活标记，并且克隆扩展，最引人注目的是 CD8+ T 细胞亚群，其中克隆性与疾病进展密切相关。有趣的是，斑块中的 T 细胞对流感和 SARS-CoV-2 等病毒表现出特异性，并与血管蛋白发生交叉反应，这提供了 T 细胞介导的自身免疫可能在机制上导致病毒相关血栓并发症的初步证据。最后，对斑块转录组的分析表明，T细胞亚群可能通过释放细胞因子双调蛋白与血管平滑肌细胞（SMC）相互作用，诱导平滑肌细胞增殖并促进不可逆斑块的形成。
Once considered primarily a disorder of lipid deposition, coronary artery disease is an incurable, life-threatening disease that is now also characterized by chronic inflammation notable for the build-up of atherosclerotic plaques containing immune cells in various states of activation and differentiation. Understanding how these immune cells contribute to disease progression may lead to the development of novel therapeutic strategies. We used single cell technology and in vitro assays to interrogate the immune microenvironment of human coronary atherosclerotic plaque at different stages of maturity. In addition to macrophages, we found a high proportion of αβ T cells in the coronary plaques. Most of these T cells lack high expression of CCR7 and L-selectin, indicating that they are primarily “antigen-experienced,” memory cells. Notably, nearly one-third of these cells express the HLA-DRA surface marker, signifying activation through their T cell-antigen receptors (TCRs). Consistent with this, TCR repertoire analysis confirmed the presence of activated αβ T cells (CD4 << CD8), exhibiting clonal expansion of specific TCRs. Interestingly, we found that these plaque T cells had TCRs specific for influenza, coronavirus, and other viral epitopes, which share sequence homologies to proteins found on smooth muscle cells and endothelial cells, suggesting potential auto-immune mediated T cell activation in the absence of active infection. To better understand the potential function of these activated plaque T cells, we then interrogated their transcriptome at the single cell level. Of the 3 T cell phenotypic clusters with the highest expression of the activation marker HLA-DRA identified by the Seurat algorithm, two clusters express a proinflammatory and cytolytic signature characteristic of CD8 cells, while the other expresses amphiregulin, which promotes smooth muscle cell proliferation and fibrosis, and, thus, contributes to plaque progression. Taken together, these findings demonstrate that plaque T cells are clonally expanded potentially by antigen engagement, are potentially reactive to self-epitopes, and may interact with smooth muscle cells and macrophages in the plaque microenvironment. Although recent studies using single cell RNA sequencing have characterized T cells within atherosclerotic plaques, these analysis have been limited to murine aortic lesions and advanced human carotid plaques. While T cell clones have been defined in the blood of patients with atherosclerosis, the plaque T cell repertoire has yet to be analyzed. Here, we provide the first comprehensive map of the human coronary plaque T cell repertoire and its transcriptome across the disease spectrum. We demonstrate that T cell effector memory clusters predominate, express activation markers, and are clonally expanded, most strikingly in the CD8+ T cell subset where clonality tracks with disease progression. Interestingly, plaque residing T cells display specificity to viruses including influenza and SARS-CoV-2 and cross-react with vascular proteins, providing preliminary evidence that T cell mediated auto-immunity may contribute mechanistically to viral-associated thrombotic complications. Finally, analysis of the plaque transcriptome reveals T cell subpopulations may interact with vascular smooth muscle cells (SMCs) through the release of the cytokine amphiregulin, inducing the proliferation of smooth muscle cells and promoting the formation of irreversible plaques.