Commensal-Specific CD4+ Cells From Patients With Crohn's Disease Have a T-Helper 17 Inflammatory Profile
Commensal-Specific CD4+ Cells From Patients With Crohn's Disease Have a T-Helper 17 Inflammatory Profile
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DOI：
10.1053/j.gastro.2016.05.050
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发表时间：
2016-09-01
期刊：
影响因子：
29.4
通讯作者：
Salas, Azucena
中科院分区：
文献类型：
--
作者：
Calderon-Gomez, Elisabeth;Bassolas-Molina, Helena;Salas, Azucena
背景与目的：克罗恩病（CD）与对肠道微生物群的免疫应答改变有关，主要是基于对微生物蛋白的血清反应性增加。虽然T细胞被认为有助于CD的发展，但对所涉及的抗原知之甚少。我们研究了从CD患者中分离的T细胞的抗原特异性。方法：我们从65例CD患者和45名健康人（对照组）中分离外周血单个核细胞。我们研究了T-细胞的反应性，使用增殖试验（基于胸苷掺入和羧基荧光素琥珀酰亚胺酯稀释）的肠道微生物抗原。使用微阵列和实时聚合酶链反应分析确定基因表达模式。通过酶联免疫吸附试验、流式细胞术或多重细胞因子试验测量细胞因子、趋化因子和抗体。从7例无炎症性肠病的患者的手术切除标本中获得肠隐窝。我们检测了肠道特异性CD 4（+）T细胞对这些样本中原代肠上皮细胞的影响。研究结果：与对照组相比，细菌蛋白FlaX、A4-fla 2和YidX增加了从CD患者外周血分离的CD 4（+）T细胞的增殖。在对照组的血液样本中，FlaX、A4-fla 2或YidX特异性的CD 4（+）T细胞具有辅助性T细胞（Th）1表型; CD患者中这些蛋白特异性的CD 4（+）T细胞中较大比例具有Th 17表型或产生Th 1和Th 17细胞因子。当从CD患者的结肠特异性CD 4（+）T细胞收集的上清液应用于健康的肠上皮细胞时，上皮细胞增加了趋化因子（C-X-C基序）配体1（CXCL 1）、CXCL 8和CC趋化因子配体20（CCL 20）的表达。结论：与对照组的T细胞相比，CD患者中更大比例的肠道特异性CD 4（+）T细胞具有Th 17表型或产生Th 1和Th 17细胞因子;这可能有助于CD患者的肠道炎症。这些细胞可能是治疗CD的靶点。来自健康个体和CD患者的骨髓特异性CD 4（+）T细胞的转录数据已保存在国家生物技术信息中心的基因表达综合库中（登录号：GSE 70469）。
BACKGROUND & AIMS: Crohn's disease (CD) has been associated with an altered immune response to commensal microbiota, mostly based on increased seroreactivity to microbial proteins. Although T cells are believed to contribute to the development of CD, little is known about the antigens involved. We investigated the antigen-specificity of T cells isolated from patients with CD. METHODS: We isolated peripheral blood mononuclear cells from 65 patients with CD and 45 healthy individuals (controls). We investigated T-cell reactivity to commensal microbial antigens using proliferation assays (based on thymidine incorporation and carboxyfluorescein succinimidyl ester dilution). Gene expression patterns were determined using microarray and real-time polymerase chain reaction analyses. Cytokines, chemokines, and antibodies were measured by enzyme-linked immunosorbent assay, flow cytometry, or multiplex cytokine assays. Intestinal crypts were obtained from surgical resection specimens of 7 individuals without inflammatory bowel disease. We examined the effects of commensal-specific CD4(+) T cells on primary intestinal epithelial cells from these samples. RESULTS: The bacterial proteins FlaX, A4-fla2, and YidX increased proliferation of CD4(+) T cells isolated from peripheral blood of patients with CD compared with controls. In blood samples from controls, CD4(+) T cells specific for FlaX, A4-fla2, or YidX had a T-helper (Th) 1 phenotype; a larger proportion of CD4(+) T cells specific for these proteins in patients with CD had a Th17 phenotype or produced Th1 and Th17 cytokines. When supernatants collected from commensal-specific CD4(+) T cells from patients with CD were applied to healthy intestinal epithelial cells, the epithelial cells increased the expression of the chemokine (C-X-C motif) ligand 1 (CXCL1), CXCL8 and the CC chemokine ligand 20 (CCL20). CONCLUSIONS: A larger proportion of commensal-specific CD4(+) T cells from patients with CD have a Th17 phenotype or produce Th1 and Th17 cytokines, compared with T cells from controls; this might contribute to intestinal inflammation in patients with CD. These cells might be targeted for treatment of CD. The transcriptional data of commensal-specific CD4(+) T cells from healthy individuals and CD patients have been deposited in the Gene Expression Omnibus at the National Center for Biotechnology Information (accession no: GSE70469).