Łuczyński W, Wawrusiewicz-Kurylonek N, Szypowska A, Iłendo E, Bossowski A, Krętowski A, Stasiak-Barmuta A
Department of Pediatrics, Endocrinology, Diabetology with Cardiology Division, Poland.
Exp Clin Endocrinol Diabetes. 2012 Feb;120(2):101-9. doi: 10.1055/s-0031-1284432. Epub 2011 Sep 15.
There is increasing evidence that T-regulatory (Treg) cells could be used to prevent or cure autoimmune diseases including type 1 diabetes mellitus (T1DM). The aim of the present study was to verify the hypothesis that functional Treg cells can be generated from conventional T-cells separated from a small amount of peripheral blood of children with newly diagnosed T1DM (N=25).
CD4(+)CD25(-) cells were cultured with Treg expander (CD3/CD28) and IL-2 for generating de novo Treg cells. The assessment of the expression of selected genes and proteins critical to Treg function and the proliferation assays were performed with the use of real-time RT-PCR and flow cytometry.
After a 4-week stimulation with Treg expander and IL-2, the percentage of T-regulatory cells was significantly higher compared to the cells treated with medium alone (with no difference between diabetic and control children). However, we found some disturbances in the gene expression at mRNA level for molecules crucial for T-reg function. The induced Tregs from diabetic and control children were fully functional as assessed in proliferation assays.
Despite some disturbances at mRNA level in the critical gene expression, the suppressive properties of induced Treg cells from diabetic and control children were effective.
越来越多的证据表明，调节性T（Treg）细胞可用于预防或治疗包括1型糖尿病（T1DM）在内的自身免疫性疾病。本研究的目的是验证从新诊断的T1DM儿童（N = 25）少量外周血中分离出的常规T细胞能够产生功能性Treg细胞这一假设。
将CD4(+)CD25(-)细胞与Treg扩增剂（CD3/CD28）和白细胞介素-2一起培养，以产生新生Treg细胞。使用实时逆转录聚合酶链反应和流式细胞术对Treg功能关键的选定基因和蛋白质表达进行评估，并进行增殖测定。
在用Treg扩增剂和白细胞介素-2刺激4周后，与仅用培养基处理的细胞相比，调节性T细胞的百分比显著更高（糖尿病儿童和对照儿童之间无差异）。然而，我们发现Treg功能关键分子的mRNA水平基因表达存在一些干扰。在增殖测定中评估发现，糖尿病儿童和对照儿童诱导产生的Tregs具有完全功能。
尽管关键基因表达在mRNA水平存在一些干扰，但糖尿病儿童和对照儿童诱导产生的Treg细胞的抑制特性是有效的。
