Vorobjova Tamara, Uibo Oivi, Heilman Kaire, Rägo Tiina, Honkanen Jarno, Vaarala Outi, Tillmann Vallo, Ojakivi Ivi, Uibo Raivo
Department of Immunology, Institute of General and Molecular Pathology, University of Tartu, Tartu, Estonia.
Scand J Gastroenterol. 2009;44(4):422-30. doi: 10.1080/00365520802624177.
To determine whether the expression of FOXP3 is changed in small-bowel mucosa in coeliac disease (CD).
The study comprised 52 patients (mean age 8.01+/-6.14 years) who had undergone small-bowel biopsies. CD only was diagnosed in 16 patients, and CD with type I diabetes mellitus (T1D) in 7. These 23 patients and 4 others without CD had partial or subtotal villous atrophy (PVA, SVA). Twenty-five persons without CD had normal mucosa. The transcription level of the FOXP3 gene (Hs00203958_m1) was evaluated in biopsy samples (small bowel) using TaqMan gene expression assays. FOXP3 protein in mucosal cells was evaluated with mouse anti-human FOXP3 antibodies and CD25(+), and CD4(+) T cells were evaluated by mouse monoclonal antibodies.
Expression of FOXP3 mRNA was higher in both PVA and SVA compared to normal mucosa (p=0.007). Patients with CD and T1D had higher expression of FOXP3 mRNA than patients with CD alone (p=0.02). The number of FOXP3(+) cells in intestinal mucosa was higher in patients with CD, especially those with coexisting T1D, than in those with normal mucosa (p=0.01). The results of double staining showed that, among all positive cells, FOXP3 expression alone was revealed in 25.6% of the cells, CD25 positivity in 18% and both markers simultaneously were found in 56.5% of lymphocytes (p=0.03). Double staining for CD4 and FOXP3 showed that 87.5% of cells were CD4(+), 2.8% were FOXP3(+) and 9.7% of cells simultaneously expressed the CD4 and FOXP3 markers.
A more pronounced expression of FOXP3 mRNA and also the number of FOXP3(+) cells (with simultaneous expression of CD25 and CD4 markers) were found in the small-bowel biopsy specimens obtained from children with CD, particularly those with coexisting T1D, compared with the FOXP3 expression in normal mucosa.
确定乳糜泻（CD）患者小肠黏膜中FOXP3的表达是否发生变化。
本研究纳入了52例接受小肠活检的患者（平均年龄8.01±6.14岁）。其中仅诊断为CD的患者有16例，合并1型糖尿病（T1D）的CD患者有7例。这23例患者以及另外4例无CD的患者存在部分或全绒毛萎缩（PVA，SVA）。25例无CD的患者黏膜正常。使用TaqMan基因表达分析方法评估活检样本（小肠）中FOXP3基因（Hs00203958_m1）的转录水平。用小鼠抗人FOXP3抗体评估黏膜细胞中的FOXP3蛋白，并用小鼠单克隆抗体评估CD25(+)和CD4(+) T细胞。
与正常黏膜相比，PVA和SVA中FOXP3 mRNA的表达均较高（p = 0.007）。合并T1D的CD患者FOXP3 mRNA的表达高于单纯CD患者（p = 0.02）。CD患者，尤其是合并T1D的患者，肠道黏膜中FOXP3(+)细胞的数量高于黏膜正常的患者（p = 0.01）。双重染色结果显示，在所有阳性细胞中，仅FOXP3表达的细胞占25.6%，CD25阳性的细胞占18%，两种标志物同时表达的淋巴细胞占56.5%（p = 0.03）。CD4和FOXP3的双重染色显示，87.5%的细胞为CD4(+)，2.8%的细胞为FOXP3(+)，9.7%的细胞同时表达CD4和FOXP3标志物。
与正常黏膜中的FOXP3表达相比，在CD患儿，尤其是合并T1D的患儿的小肠活检标本中发现FOXP3 mRNA的表达更明显，且FOXP3(+)细胞数量更多（同时表达CD25和CD4标志物）。
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