Chrul Sławomir, Polakowska Ewa, Mycko Marcin, Fendler Wojciech, Zwiech Rafał, Szadkowska Agnieszka
Department of Kidney Transplantation, Medical University of Lodz, Poland.
Pediatr Endocrinol Diabetes Metab. 2013;19(3):91-5.
The immunologic reaction of pancreatic islets destruction leads to the occurrence of type 1 diabetes mellitus (T1D). The autoreactive lymphocytes play the pivotal role in this process although mechanisms regulating the lymphocyte migration and infiltration of Langerhans islets have not been fully understood yet. The in vitro studies showed natural killer (NK) cells potency to initiate pancreatic islets cell lyses. Many authors postulate that NK cells may be involved in this reaction.
The aim of the study was to evaluate the effect of IL-2, IL-12 and IL-15 stimulation on peripheral blood NK cells in children suffering from type 1 diabetes mellitus in comparison to healthy controls.
Fifteen children with type 1 diabetes and 10 healthy adults were examined. NK cells were isolated by the magnetic cell separation system (MACS). For activation, NK cells were cultured with IL-2, IL-12 and IL-15 for 24 hours. The production of IFN-γ and IL-10 by NK cells was measured using commercial ELISA kits. FACS analysis of cell surface antigens--CD16, CD56, NKG2D and CD137 was performed using LSR II flow cytometer.
In children with T1D the IFN-γ median concentration in supernatant obtained from NK cells culture was 16.831 ng/ml (inter quartile range 5.566-25.509) and did not statistically differ from median IFN-γ concentration in the control group--14.810 ng/ml (7.022-18.785), p = 0.76. In contrast, the IL-10 median concentration was statistically higher in T1D patients 7.87 pg/ml (1.32-11.37) than in healthy participants--1.41 pg/ml (1.05-4.81), p = 0.01. The median (inter-quartile range) percentage of NK NKG2D(+) was found in 0.42% (0.28-0.76) cells of TID patients versus 0.72% (0.53-1.08) in the controls (p = 0.05). There was no difference between -T1D group and the control group in regard to NK cells expressing CD137 - 6.58% (3.38-12.4) versus 6.85% (2.94-10.8); p = 0.8.
The observed activity of NK cells after in vitro stimulation by IL2, IL-12 and IL15 in children suffering from type 1 diabetes mellitus indicates the tendency for supporting the inhibition of autoimmunological reaction by increased IL10 synthesis and increased number of NK cells with surface NKG2D receptors.
胰岛破坏的免疫反应导致1型糖尿病（T1D）的发生。自身反应性淋巴细胞在这一过程中起关键作用，尽管调节淋巴细胞迁移和朗格汉斯胰岛浸润的机制尚未完全明确。体外研究显示自然杀伤（NK）细胞有引发胰岛细胞溶解的能力。许多作者推测NK细胞可能参与了这一反应。
本研究旨在评估与健康对照相比，白细胞介素-2（IL-2）、白细胞介素-12（IL-12）和白细胞介素-15（IL-15）刺激对1型糖尿病儿童外周血NK细胞的影响。
对15名1型糖尿病儿童和10名健康成年人进行检查。通过磁珠细胞分选系统（MACS）分离NK细胞。为进行激活，将NK细胞与IL-2、IL-12和IL-15培养24小时。使用商用酶联免疫吸附测定（ELISA）试剂盒检测NK细胞产生的干扰素-γ（IFN-γ）和白细胞介素-10（IL-10）。使用LSR II流式细胞仪对细胞表面抗原——CD16、CD56、自然杀伤细胞2族成员D（NKG2D）和CD137进行流式细胞术分析。
在1型糖尿病儿童中，NK细胞培养上清液中IFN-γ的中位数浓度为16.831纳克/毫升（四分位间距5.566 - 25.509），与对照组中IFN-γ的中位数浓度14.810纳克/毫升（7.022 - 18.785）相比，差异无统计学意义，p = 0.76。相比之下，1型糖尿病患者中IL-10的中位数浓度在统计学上高于健康参与者，分别为7.87皮克/毫升（1.32 - 11.37）和1.41皮克/毫升（1.05 - 4.81），p = 0.01。1型糖尿病患者NK细胞中NKG2D(+)的中位数（四分位间距）百分比在0.42%（0.28 - 0.76）的细胞中，而对照组为0.72%（0.53 - 1.08）（p = 列的中位数（四分位间距）百分比在0.42%（0.28 - 0.76）的细胞中，而对照组为0.72%（0.53 - 1.08）（p = 0.05）。在表达CD137的NK细胞方面——1型糖尿病组与对照组之间无差异，分别为6.58%（3.38 - 12.4）与6.85%（2.94 - 10.8）；p = 0.8。
在体外经IL-2、IL-12和IL-15刺激后，观察到1型糖尿病儿童NK细胞的活性，这表明其有通过增加IL-10合成以及增加具有表面NKG2D受体的NK细胞数量来支持抑制自身免疫反应的趋势。
