Increased FOXP3 expression in small-bowel mucosa of children with coeliac disease and type I diabetes mellitus

Abstract

Objective. To determine whether the expression of FOXP3 is changed in small-bowel mucosa in coeliac disease (CD). Material and methods. 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. Results. 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. Conclusions. 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.

• CD4
• CD25
• children
• coeliac disease
• FOXP3
• immunohistochemistry
• real-time PCR
• Treg cells
• type I diabetes