Brynychova Iva, Hoffmanova Iva, Dvorak Milos, Dankova Pavlina
Department of Anthropology and Human Genetics, Faculty of Science, Charles University in Prague, Czech Republic.
Second Department of Internal Medicine, Third Faculty of Medicine, Charles University in Prague, Czech Republic.
Adv Clin Exp Med. 2016 Sep-Oct;25(5):887-893. doi: 10.17219/acem/39940.
Celiac disease (CD) is an organ-specific autoimmune disease, and both adaptive and innate immunity are involved in its development.
The aim of the study was to determine whether the markers of intestinal mucosal inflammation in CD can be detected in peripheral blood monocytes (PBMs), and whether the immune properties of PBMs change as the clinical signs and symptoms of CD improve after the introduction of a gluten-free diet (GFD). The focus was on changes in mRNA expression of selected toll-like receptors (TLR2, TLR4, TLR7), stress cytokine prolactin (PRL), and proand anti-inflammatory cytokines (TNF-α, IL-6, IL-12, IL-10) in PBMs.
The study involved 20 CD patients diagnosed according to the European Society for Pediatric Gastroenterology, Hepatology and Nutrition criteria and Marsh criteria: 10 recently-diagnosed cases (rCD) and 10 on a GFD for a minimum of one year. The control group comprised 10 ageand sex-matched healthy volunteers. PBMs from peripheral blood specimens were separated using immunomagnetic CD14+ beads. Total RNA was isolated using a standard commercial kit. Cytokine and TLR mRNA levels were quantified by relative qPCR with PGK1 as a reference gene.
Significantly higher expression of TLR4 and TLR7 mRNA was observed in PBMs from rCD patients compared to the healthy controls (1.63 times higher; p < 0.05). TLR7 mRNA levels in rCDs were also significantly elevated in comparison to the CD-GFD patients (2.11 times higher; p < 0.01). TNF-α mRNA expression tended to be higher in both groups of patients; by contrast, in IL-6 mRNA, a trend to a fourfold decrease was detected in PBMs from the CD-GFD subjects. IL-10, IL-12 and PRL levels did not differ among the groups.
The data suggest that the inflammatory process in rCD intestinal mucosa and submucosa reflecting enterocyte damage can be detected in PBMs in peripheral blood. Further, the cytokine and TLR expression profile in PBMs alters after one year of GFD treatment.
乳糜泻（CD）是一种器官特异性自身免疫性疾病，适应性免疫和先天性免疫均参与其发病过程。
本研究旨在确定CD患者肠道黏膜炎症标志物是否可在外周血单核细胞（PBM）中检测到，以及引入无麸质饮食（GFD）后，随着CD临床症状和体征改善，PBM的免疫特性是否发生变化。重点关注PBM中所选Toll样受体（TLR2、TLR4、TLR7）、应激细胞因子催乳素（PRL）以及促炎和抗炎细胞因子（TNF-α、IL-6、IL-12、IL-10）的mRNA表达变化。
本研究纳入20例根据欧洲儿科胃肠病学、肝病学和营养学会标准及马什标准确诊的CD患者：10例新诊断病例（rCD）和10例接受GFD至少一年的患者。对照组包括10名年龄和性别匹配的健康志愿者。使用免疫磁珠CD14+从外周血标本中分离PBM。使用标准商业试剂盒分离总RNA。以PGK1作为参考基因，通过相对定量PCR对细胞因子和TLR的mRNA水平进行定量。
与健康对照组相比，rCD患者PBM中TLR4和TLR7 mRNA的表达显著更高（高1.63倍；p < 0.05）。与CD-GFD患者相比，rCD患者的TLR7 mRNA水平也显著升高（高2.11倍；p < 0.01）。两组患者中TNF-α mRNA表达均有升高趋势；相比之下，在CD-GFD患者的PBM中，IL-6 mRNA有下降四倍的趋势。各组间IL-10、IL-12和PRL水平无差异。
数据表明，外周血PBM中可检测到反映rCD肠黏膜和黏膜下层肠上皮细胞损伤的炎症过程。此外，GFD治疗一年后，PBM中的细胞因子和TLR表达谱发生改变。
