Intestinal transplant (ITx) rejection is associated with memory T helper type 17 cell (Th17) infiltration of grafted tissues. Modulation of Th17 effector cell response is facilitated by T regulatory (Treg) cells, but a phenotypic characterization of this process is lacking in the context of allograft rejection. Flow cytometry was performed to examine the expression of surface receptors, cytokines, and transcription factors in Th17 and Treg cells in ITx control (n = 34) and rejection patients (n = 23). To elucidate key pathways guiding the rejection biology, we utilized RNA sequencing (RNAseq) and assessed epigenetic stability through pyrosequencing of the Treg-specific demethylated region (TSDR). We found that intestinal allograft rejection is characterized by Treg cellular infiltrates, which are polarized toward Th17-type chemokine receptor, ROR-γt transcription factor expression, and cytokine production. These Treg cell subsets have maintained epigenetic stability, as defined by FoxP3-TSDR methylation status, but displayed upregulation of functional Treg and purinergic signaling genes by RNAseq analysis such as CD39, in keeping with suppressor Th17 properties. We show that ITx rejection is associated with increased polarized cells that express a Th17-like phenotype concurrent with regulatory purinergic markers.

中文翻译：

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患有同种异体移植排斥的肠移植受者中抑制 T 辅助细胞 17 型细胞反应

肠移植 (ITx) 排斥与移植组织的记忆 T 辅助细胞 17 型细胞 (Th17) 浸润有关。Th17 效应细胞反应的调节是由调节性 T (Treg) 细胞促进的，但在同种异体移植排斥的情况下缺乏该过程的表型特征。采用流式细胞术检测 ITx 对照 (n = 34) 和排斥反应患者 (n = 23) 中 Th17 和 Treg 细胞的表面受体、细胞因子和转录因子的表达。为了阐明指导排斥生物学的关键途径，我们利用 RNA 测序 (RNAseq) 并通过 Treg 特异性去甲基化区域 (TSDR) 的焦磷酸测序评估表观遗传稳定性。我们发现肠道同种异体移植排斥的特点是 Treg 细胞浸润，这些细胞向 Th17 型趋化因子受体、ROR-γt 转录因子表达和细胞因子产生极化。这些 Treg 细胞亚群保持了由 FoxP3-TSDR 甲基化状态定义的表观遗传稳定性，但通过 RNAseq 分析（如 CD39）显示功能性 Treg 和嘌呤信号基因的上调，与抑制性 Th17 特性保持一致。我们发现 ITx 排斥与表达 Th17 样表型和调节性嘌呤能标记物的极化细胞增加有关。