Characterization of T-cell Receptor Repertoire in Inflamed Tissues of Patients with Crohn's Disease Through Deep Sequencing
Characterization of T-cell Receptor Repertoire in Inflamed Tissues of Patients with Crohn's Disease Through Deep Sequencing
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DOI：
10.1097/mib.0000000000000752
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发表时间：
2016-06-01
影响因子：
4.9
通讯作者：
Kiyotani, Kazuma
中科院分区：
文献类型：
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作者：
Chapman, Christopher G.;Yamaguchi, Rui;Kiyotani, Kazuma
背景：克罗恩病（CD）患者的肠组织中含有大量的T细胞，这些T细胞被认为是介导炎症的细胞。我们进行了详细的表征，这些T细胞repertoires.Methods：我们获得了活检组织从12例患者的新末端回肠进行评估术后复发CD和4个人与正常末端回肠，没有炎症性肠病的历史（对照）。从5例接受狭窄或穿透CD手术的患者中获得病变末端回肠样本。从组织和外周血单核细胞中提取总RNA，并产生cDNA。我们使用下一代测序来表征T细胞受体（TCR）和TCR-cDNA在回肠粘膜组织和匹配的外周血单个核细胞的17例CD，以确定寡克隆扩增的T细胞群体与CD。结果：TCR的多样性在粘膜组织中显着低于匹配的外周血单个核细胞，表明某些T细胞群体在发炎的肠组织中的扩张。在12例复发性CD患者中的4例的新末端回肠中，单一TCR克隆型CASSWTNGEQYF（TRBV 10 -1-TRBJ 2 -7）以7.0%至28.9%的频率富集。该克隆型的丰度与疾病复发的严重程度显着相关，根据Rutgeillance评分（P = 0.015）。结论：特定的T细胞群扩大在CD患者的炎症肠粘膜，其丰度与疾病复发的严重程度相关。对这些T细胞的研究可以提供关于CD发病机制的信息。深度TCR测序是一种强大的工具，可快速提供对T细胞库的深入、实时评估。
Background:Intestinal tissues of patients with Crohn's disease (CD) contain expanded populations of T cells which are believed to mediate inflammation. We performed a detailed characterization of these T-cell repertoires.Methods:We obtained biopsies from the neoterminal ileum of 12 patients undergoing evaluation for postoperative recurrent CD and 4 individuals with normal terminal ileum and no history of inflammatory bowel disease (controls). Samples of diseased terminal ileum were obtained from 5 patients undergoing surgery for stricturing or penetrating CD. Total RNA was extracted from tissues and peripheral blood mononuclear cells, and cDNAs were generated. We used next-generation sequencing to characterize T-cell receptor (TCR)- and TCR- cDNAs in ileal mucosal tissue and matched peripheral blood mononuclear cells of 17 patients with CD to identify oligoclonal expansions of T-cell populations associated with CD.Results:TCR diversity in mucosal tissue was significantly lower than that of matched peripheral blood mononuclear cells, indicating expansion of certain T-cell populations in inflamed intestinal tissue. A single TCR- clonotype, CASSWTNGEQYF (TRBV10-1-TRBJ2-7), was enriched at a frequency of 7.0% to 28.9% in the neoterminal ileum of 4 of 12 patients with recurrent CD. The abundance of this clonotype significantly correlated with the severity of disease recurrence, based on Rutgeerts score (P = 0.015).Conclusions:Specific populations of T cells are expanded in the inflamed intestinal mucosa of patients with CD; their abundance correlates with severity of disease recurrence. Studies of these T cells could provide information about mechanisms of CD pathogenesis. Deep TCR sequencing is a powerful tool that rapidly provides in-depth, real-time assessment of the T-cell repertoire.