Vrabelova Zuzana, Kolouskova Stanislava, Böhmova Kristyna, Faresjö Maria Karlsson, Sumnik Zdenek, Pechova Marta, Kverka Miloslav, Chudoba Daniel, Zacharovova Klara, Stadlerova Gabriela, Pithova Pavlina, Hladikova Marie, Stechova Katerina
Department of Paediatrics, 2nd Medical Faculty of Charles University, Prague, The Czech Republic.
Pediatr Diabetes. 2007 Oct;8(5):252-60. doi: 10.1111/j.1399-5448.2007.00308.x.
Autoreactive T cells have a crucial role in type 1 diabetes (T1D) pathogenesis.
The aim of our study was to monitor the in vitro production of cytokines by peripheral blood mononuclear cells (PBMCs) after stimulation with diabetogenic autoantigens.
Ten T1D patients (tested at the time of diagnosis and 6 and 12 months later), 10 first-degree relatives of the T1D patients, and 10 controls underwent the study.
PBMCs were stimulated with glutamic acid decarboxylase 65 (GAD65) amino acids (a.a.) 247-279, 509-528, and 524-543; proinsulin a.a. 9-23; and tyrosine phosphatase (islet antigen-2)/R2 a.a. 853-872. Interleukin (IL)-2, IL-4, IL-5, IL-6, IL-10, IL-13, interferon (IFN)-gamma, tumor necrosis factor beta, transforming growth factor beta1, and granulocyte colony-stimulating factor (GCSF) were analyzed by protein microarray.
Differences in cytokine(s) poststimulatory and mainly in basal production were observed in all groups. The most prominent findings were in controls, the higher basal levels of IL-2, IL-4, IL-5, IL-13, and GCSF were observed when compared with relatives (p < 0.05, for all). After stimulation in controls, there was a significant decrease in IL-2, IL-13, GCSF, and IFN-gamma (p < 0.05, for all). The group of relatives was the most variable in poststimulatory production. A strong correlation between cytokines production was found but groups differed in this aspect.
By multiplex analysis, it may be possible, for example, to define the risk immunological response pattern among relatives or to monitor the immune response in patients on immune modulation therapy.
自身反应性T细胞在1型糖尿病（T1D）发病机制中起关键作用。
我们研究的目的是监测外周血单个核细胞（PBMCs）在用致糖尿病自身抗原刺激后细胞因子的体外产生情况。
10名T1D患者（在诊断时、6个月和12个月后进行检测）、10名T1D患者的一级亲属以及10名对照者参与了该研究。
用谷氨酸脱羧酶65（GAD65）氨基酸（a.a.）247 - 279段、509 - 528段和524 - 543段；胰岛素原a.a. 9 - 23段；以及酪氨酸磷酸酶（胰岛抗原-2）/R2 a.a. 853 - 872段刺激PBMCs。通过蛋白质微阵列分析白细胞介素（IL）-2、IL-4、IL-5、IL-6、IL-10、IL-13、干扰素（IFN）-γ、肿瘤坏死因子β、转化生长因子β1和粒细胞集落刺激因子（GCSF）。
在所有组中均观察到刺激后细胞因子的差异，主要是基础产生水平的差异。最显著的发现是在对照组中，与亲属相比，观察到IL-2、IL-4、IL-5、IL-13和GCSF的基础水平更高（所有p < 0.05）。在对照组中刺激后，IL-2、IL-13、GCSF和IFN-γ有显著下降（所有p < 0.05）。亲属组在刺激后产生情况方面变化最大。发现细胞因子产生之间存在强相关性，但各组在这方面存在差异。
例如，通过多重分析，有可能确定亲属中的风险免疫反应模式或监测接受免疫调节治疗患者的免疫反应。
