The mRNA expression of pro- and anti-inflammatory cytokines in T regulatory cells in children with type 1 diabetes

Folia Histochem Cytobiol. 2010 Jan 1;48(1):93-100. doi: 10.2478/v10042-008-0113-5.


Type 1 diabetes mellitus (T1DM) is caused by the autoimmune-mediated destruction of insulin-producing beta cells in the pancreas. T regulatory cells (Tregs) represent an active mechanism of suppressing autoreactive T cells that escape central tolerance. The aim of our study was to test the hypothesis that T regulatory cells express pro- and anti-inflammatory cytokines, elements of cytotoxicity and OX40/4-1BB molecules. The examined group consisted of 50 children with T1DM. Fifty two healthy individuals (control group) were enrolled into the study. A flow cytometric analysis of T-cell subpopulations was performed using the following markers: anti-CD3, anti-CD4, anti-CD25, anti-CD127, anti-CD134 and anti-CD137. Concurrently with the flow cytometric assessment of Tregs we separated CD4+CD25+CD127dim/- cells for further mRNA analysis. mRNA levels for transcription factor FoxP3, pro- and anti-inflammatory cytokines (interferon gamma, interleukin-2, interleukin-4, interleukin-10, transforming growth factor beta1 and tumor necrosis factor alpha), activatory molecules (OX40, 4-1BB) and elements of cytotoxicity (granzyme B, perforin 1) were determined by real-time PCR technique. We found no alterations in the frequency of CD4+CD25highCD127low cells between diabetic and control children. Treg cells expressed mRNA for pro- and anti-inflammatory cytokines. Lower OX40 and higher 4-1BB mRNA but not protein levels in Treg cells in diabetic patients compared to the healthy children were noted. Our observations confirm the presence of mRNA for pro- and anti-inflammatory cytokines in CD4+CD25+CD127dim/- cells in the peripheral blood of children with T1DM. Further studies with the goal of developing new strategies to potentiate Treg function in autoimmune diseases are warranted.

MeSH terms

  • Case-Control Studies
  • Child
  • Cytokines / genetics*
  • Cytokines / metabolism
  • Diabetes Mellitus, Type 1 / genetics*
  • Diabetes Mellitus, Type 1 / immunology*
  • Flow Cytometry
  • Gene Expression Regulation*
  • Health
  • Humans
  • Inflammation Mediators / metabolism*
  • Interleukin-2 Receptor alpha Subunit / metabolism
  • Interleukin-7 Receptor alpha Subunit / metabolism
  • Lymphocyte Subsets / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Receptors, OX40 / genetics
  • Receptors, OX40 / metabolism
  • T-Lymphocytes, Regulatory / metabolism*
  • Tumor Necrosis Factor Receptor Superfamily, Member 9 / genetics
  • Tumor Necrosis Factor Receptor Superfamily, Member 9 / metabolism


  • Cytokines
  • Inflammation Mediators
  • Interleukin-2 Receptor alpha Subunit
  • Interleukin-7 Receptor alpha Subunit
  • RNA, Messenger
  • Receptors, OX40
  • TNFRSF4 protein, human
  • TNFRSF9 protein, human
  • Tumor Necrosis Factor Receptor Superfamily, Member 9