Lately, in two elegant studies with the use of flow cytometry and real-time PCR, investigators demonstrated that T regulatory cells can be separated with the combination of CD4, CD25 and

CD127 (IL-7R) [19, 20]. At the beginning of our experiment, we also tested the correlation between low expression of CD127 and expression of transcription factor FoxP3. In accordance to Seddiki et al. and Liu et al., we observed that most of the CD127low/− cells were FoxP3 positive, and the correlation between CD127low/− and FoxP3+ PLX4032 price cells was very high [19, 20]. These results allowed us to regard CD4+CD25+ CD127low/− cells as Tregs and separate them for further studies at mRNA level. In previous experiments conducted by other authors, CD4+CD25+ subpopulation

was used for the assessment of mRNA expression in T regulatory cells [21]. For more precise results, we used newly developed kit for separating CD4+CD25+CD127dim/− cells, but the high purity of isolation selleck chemicals was very difficult to achieve, and the amounts of separated cells were relatively small: 104–105. However, the real-time PCR technique allows for the assessment of mRNA for many genes in one, small sample. As mentioned previously, there are no reports concerning T regulatory cells in patients with MS neither in children nor in adults. Several studies indicated the association between elevated total white blood cell/lymphocyte numbers and components of MS [22]. In another analysis, the number of CD4+ cells correlated with components of MS [23]. This correlation was not confirmed in our group. To date, Tideglusib only one report concerned Tregs in obese children. Svec et al., in accordance with our results, did not find any differences in the percentage of CD4+CD25highFoxP3+ cells between obese and non-obese children. However, the study groups were

very small (12 versus 10) [14]. Classically, it was believed that Tregs act via contact-dependent, cytokine-independent manner; however, the most recent data suggest the involvement of some cytokines including IL-35 and IL-10 in this process [24]. Thus, as suggested by Kryczek et al. [25], we used the combination of FoxP3 expression and cytokine profile for Tregs evaluation. Our results from gene expression analysis can suggest the dysfunction of T regulatory cells in children with MS. Although the FoxP3 expression was not altered, we noted lower mRNA amounts for genes encoding cytokines from IL-12 family, including IL-12A, IL-27 and IL-35 (Ebi3). Despite similar composition, the activity of those cytokines is quite different (discussed in [26]). IL-12 plays a significant role in autoimmune disorders.