IL-2和IL-7对优化检测1型糖尿病患者谷氨酸脱羧酶65特异性T细胞反应的比较

发布时间：2018-05-22 09:08

本文选题：型糖尿病 + T细胞　；参考：《中南大学学报(医学版)》2017年11期

【摘要】：目的:探讨何种细胞因子(IL-2或IL-7)及其最佳浓度能够最有效地改善酶联免疫斑点法(enzyme-linked immunospot,ELISPOT)检测1型糖尿病患者谷氨酸脱羧酶65(glutamic acid decarboxylase 65,GAD65)特异性T细胞反应的效率。方法:选取20例1型糖尿病患者(A组)及年龄和性别均匹配的16例正常对照者(B组);Ficoll法分离外周血单个核细胞(peripheral blood mononuclear cells,PBMCs),以人GAD65、内对照、巴斯德五合一疫苗为抗原,加入不同浓度的IL-2[0 U/m L(1组)、0.5 U/m L(2组)、2.5 U/m L(3组)和12.5 U/m L(4组)],ELISPOT检测上述各浓度组分泌干扰素-γ(interferon-gamma,IFN-γ)的CD4+T细胞,比较各浓度组的GAD65孔(信号)、内对照孔(背景)的斑点数、斑点净值以及刺激指数(即信噪比,stimulating index,SI);另选取21例1型糖尿病患者(C组)及12例正常对照者(D组),检测针对GAD65抗原的特异性T细胞反应,比较联合IL-2(2.5 U/m L,上述实验已证明其为最优浓度,5组)与联合IL-7(0.5 ng/m L,6组)所产的斑点数和SI。结果:1)A组在加入IL-2刺激后,各浓度组中GAD65反应性T细胞数均较A1组增高,而各浓度下的内对照孔的斑点数亦呈比例升高。各浓度组斑点净值比较显示:A4组与A3组差异无统计学意义(P0.05),而SI的比较则提示A3组最高;且仅A3组的SI(2.8)高于B3组(1.5),差异有统计学意义(P0.05)。2)C6组及D6组的GAD65孔斑点数均分别略高于C5组及D5组,且C6组及D6组的斑点数增幅也分别高于C5组及D5组,但差异均无统计学意义(P0.05)。C5组的斑点净值(5.5)和SI(2.8)均显著高于C6组的斑点净值(4.3)和SI(1.8),差异均有统计学意义(均P0.05)。结论:2.5 U/m L为ELISPOT检测1型糖尿病患者GAD65特异性T细胞反应的IL-2最佳浓度;IL-2较IL-7更有利于改善ELISPOT检测的SI。
[Abstract]:Objective: To investigate the efficiency of the cytokine (IL-2 or IL-7) and its optimum concentration to improve the efficiency of enzyme-linked immunospot (ELISPOT) for detecting the specific cell reaction of glutamic decarboxylase 65 (glutamic acid decarboxylase 65, GAD65) in type 1 diabetic patients. Methods: 20 patients with type 1 diabetes mellitus (A group) were selected. 16 normal controls (group B) were matched with age and sex; Ficoll method was used to separate peripheral blood mononuclear cells (peripheral blood mononuclear cells, PBMCs), with human GAD65, internal control, and Pasteur five one vaccine as antigen, adding different concentrations of IL-2[0 U/m L (1 groups), 0.5 U/m (2 groups), 2.5 3 groups, 4 groups (4 groups)] The CD4+T cells that secrete interferon - gamma (interferon-gamma, IFN- gamma) in the above concentration groups were measured to compare the number of GAD65 holes (signals) in each concentration group, the number of spots in the internal control hole (background), the net spot and the stimulus index (signal-to-noise ratio, stimulating index, SI), and the other 21 cases of type 1 diabetes mellitus (C group) and 12 normal controls (D group), which were detected for GA. The specific T cell response of D65 antigen was compared with the combined IL-2 (2.5 U/m L, which proved that it was the best concentration, 5 groups) and the number of spots and SI. produced by the combined IL-7 (0.5 ng/m L, 6 groups) and SI. results: 1) the number of GAD65 reactive cells in each concentration group was higher than that in the IL-2 stimulation group, and the number of spots in the inner control holes at each concentration were also The comparison of net spots in each concentration group showed that there was no significant difference between the A4 group and the A3 group (P0.05), while the comparison of the SI showed that the A3 group was the highest, and the SI (2.8) of the A3 group was higher than that of the B3 group (1.5), and the difference was statistically significant (P0.05).2) and the number of spots in the C6 group and the D6 group were slightly higher than those in the group and the group. The growth rate was higher than that of group C5 and D5 group, but the difference was not statistically significant (P0.05), the net spot net value of.C5 group (5.5) and SI (2.8) were significantly higher than that of C6 group (4.3) and SI (1.8). The difference was statistically significant (P0.05). Conclusion: 2.5 U/m L is the best concentration of GAD65 specific cell reaction in type 1 diabetic patients. IL-7 is more beneficial to the improvement of SI. for ELISPOT detection
【作者单位】：南昌市第三医院代谢内分泌科;中南大学湘雅二医院代谢内分泌研究所中南大学糖尿病中心糖尿病免疫学教育部重点实验室国家代谢性疾病临床医学研究中心;
【基金】：国家自然科学基金(30400217)~~
【分类号】：R587.1

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