發(fā)布時(shí)間：2018-05-22 09:08

  本文選題：型糖尿病 + T細(xì)胞��； 參考：《中南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)》2017年11期

【摘要】：目的:探討何種細(xì)胞因子(IL-2或IL-7)及其最佳濃度能夠最有效地改善酶聯(lián)免疫斑點(diǎn)法(enzyme-linked immunospot,ELISPOT)檢測(cè)1型糖尿病患者谷氨酸脫羧酶65(glutamic acid decarboxylase 65,GAD65)特異性T細(xì)胞反應(yīng)的效率。方法:選取20例1型糖尿病患者(A組)及年齡和性別均匹配的16例正常對(duì)照者(B組);Ficoll法分離外周血單個(gè)核細(xì)胞(peripheral blood mononuclear cells,PBMCs),以人GAD65、內(nèi)對(duì)照、巴斯德五合一疫苗為抗原,加入不同濃度的IL-2[0 U/m L(1組)、0.5 U/m L(2組)、2.5 U/m L(3組)和12.5 U/m L(4組)],ELISPOT檢測(cè)上述各濃度組分泌干擾素-γ(interferon-gamma,IFN-γ)的CD4+T細(xì)胞,比較各濃度組的GAD65孔(信號(hào))、內(nèi)對(duì)照孔(背景)的斑點(diǎn)數(shù)、斑點(diǎn)凈值以及刺激指數(shù)(即信噪比,stimulating index,SI);另選取21例1型糖尿病患者(C組)及12例正常對(duì)照者(D組),檢測(cè)針對(duì)GAD65抗原的特異性T細(xì)胞反應(yīng),比較聯(lián)合IL-2(2.5 U/m L,上述實(shí)驗(yàn)已證明其為最優(yōu)濃度,5組)與聯(lián)合IL-7(0.5 ng/m L,6組)所產(chǎn)的斑點(diǎn)數(shù)和SI。結(jié)果:1)A組在加入IL-2刺激后,各濃度組中GAD65反應(yīng)性T細(xì)胞數(shù)均較A1組增高,而各濃度下的內(nèi)對(duì)照孔的斑點(diǎn)數(shù)亦呈比例升高。各濃度組斑點(diǎn)凈值比較顯示:A4組與A3組差異無統(tǒng)計(jì)學(xué)意義(P0.05),而SI的比較則提示A3組最高;且僅A3組的SI(2.8)高于B3組(1.5),差異有統(tǒng)計(jì)學(xué)意義(P0.05)。2)C6組及D6組的GAD65孔斑點(diǎn)數(shù)均分別略高于C5組及D5組,且C6組及D6組的斑點(diǎn)數(shù)增幅也分別高于C5組及D5組,但差異均無統(tǒng)計(jì)學(xué)意義(P0.05)。C5組的斑點(diǎn)凈值(5.5)和SI(2.8)均顯著高于C6組的斑點(diǎn)凈值(4.3)和SI(1.8),差異均有統(tǒng)計(jì)學(xué)意義(均P0.05)。結(jié)論:2.5 U/m L為ELISPOT檢測(cè)1型糖尿病患者GAD65特異性T細(xì)胞反應(yīng)的IL-2最佳濃度;IL-2較IL-7更有利于改善ELISPOT檢測(cè)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
【作者單位】： 南昌市第三醫(yī)院代謝內(nèi)分泌科;中南大學(xué)湘雅二醫(yī)院代謝內(nèi)分泌研究所中南大學(xué)糖尿病中心糖尿病免疫學(xué)教育部重點(diǎn)實(shí)驗(yàn)室國(guó)家代謝性疾病臨床醫(yī)學(xué)研究中心;
【基金】：國(guó)家自然科學(xué)基金(30400217)~~
【分類號(hào)】：R587.1

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