Autoreactive T cells may contribute to post-viral myocarditis induced with Coxsackievirus B3 (CVB3), but the underlying mechanisms of their generation are unclear. Here, we have comprehensively analyzed the generation of antigen-specific, autoreactive T cells in the mouse model of CVB3 infection for antigens implicated in patients with myocarditis/dilated cardiomyopathy. First, comparative analysis of CVB3 proteome with five autoantigens led us to identify three mimicry epitopes, one each from adenine nucleotide translocator 1 (ANT), sarcoplasmic/endoplasmic reticulum Ca2+ ATPase 2a (SERCA2a) and cardiac troponin I. None of these induced cross-reactive T cell responses. Next, we generated major histocompatibility complex (MHC) class II dextramers to enumerate the frequencies of antigen-specific T cells to determine whether T cells with multiple antigen specificities are generated by CVB3 infection. These analyses revealed appearance of CD4 T cells positive for SERCA2a 971-990, and cardiac myosin heavy chain-α (Myhc) 334-352 dextramers, both in the periphery and also in the hearts of CVB3-infected animals. While ANT 21-40 dextramer+ T cells were inconsistently detected, the β1-adrenergic receptor 181-200/211-230 or branched chain α-ketoacid dehydrogenase kinase 111-130 dextramer+ cells were absent. Interestingly, SERCA2a 971-990, Myhc 334-352 and ANT 21-40 dextramer+ cells were also detected in the liver indicating that they may have a pathogenic role. Finally, we demonstrate that the SERCA2a 971-990-reactive T cells generated in CVB3 infection could transfer disease to naïve mice. The data suggest that CVB3 infection can lead to the generation of autoreactive T cells for multiple antigens indicating a possibility that the autoreactive T cells localized in the liver can potentially circulate and contribute to the development of viral myocarditis.

中文翻译：

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病毒性心肌炎涉及在 CVB3 感染小鼠模型中产生具有多种抗原特异性的自身反应性 T 细胞，这些抗原定位于淋巴和非淋巴器官。

自身反应性 T 细胞可能导致柯萨奇病毒 B3 (CVB3) 诱导的病毒后心肌炎，但其产生的潜在机制尚不清楚。在这里，我们全面分析了在 CVB3 感染小鼠模型中产生的抗原特异性、自身反应性 T 细胞，用于与心肌炎/扩张型心肌病患者相关的抗原。首先，通过对 CVB3 蛋白质组与五种自身抗原的比较分析，我们确定了三个模拟表位，其中一个来自腺嘌呤核苷酸易位蛋白 1 (ANT)、肌浆/内质网 Ca2+ ATPase 2a (SERCA2a) 和心肌肌钙蛋白 I。这些都没有诱导交叉反应性 T 细胞反应。下一个，我们生成了主要组织相容性复合体 (MHC) II 类右旋分子，以计算抗原特异性 T 细胞的频率，以确定具有多种抗原特异性的 T 细胞是否由 CVB3 感染产生。这些分析显示，在 CVB3 感染动物的外周和心脏中，出现了 SERCA2a 971-990 和心脏肌球蛋白重链-α (Myhc) 334-352 右旋体阳性的 CD4 T 细胞。虽然 ANT 21-40 右旋糖体 + T 细胞的检测不一致，但不存在 β1-肾上腺素能受体 181-200/211-230 或支链 α-酮酸脱氢酶激酶 111-130 右旋体 + 细胞。有趣的是，在肝脏中也检测到 SERCA2a 971-990、Myhc 334-352 和 ANT 21-40 右旋糖体+细胞，表明它们可能具有致病作用。最后，我们证明在 CVB3 感染中产生的 SERCA2a 971-990 反应性 T 细胞可以将疾病转移给幼稚小鼠。数据表明，CVB3 感染可导致产生多种抗原的自身反应性 T 细胞，这表明位于肝脏中的自身反应性 T 细胞可能循环并促进病毒性心肌炎的发展。