胰岛素B9-23多肽对树突状细胞表型及功能的影响  

Impact of insulin B9-23 peptide on phenotype and function of dendritic cells

作　　者：曹维[1] 季琝君[2] 邢春燕[1] 杨涛[1] 陈钰[3] 刘煜[1,4] 

机构地区：[1]南京医科大学第一附属医院内分泌科,江苏南京210029 [2]南京医科大学病原生物学系,江苏南京210029 [3]吉林大学白求恩医学院病原生物学教研究,吉林长春130021 [4]吉林大学第二医院内分泌科,吉林长春130012

出　　处：《南京医科大学学报（自然科学版）》2010年第7期914-918,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省自然科学基金资助(BK2006531)

摘　　要：目的:观察小鼠骨髓来源的树突状细胞(DCs)负载胰岛素B9-23多肽后其表型和功能的变化,探讨其对免疫状态的影响。方法:①应用10ng/ml的粒细胞巨噬细胞集落刺激因子(GM-CSF)及10ng/ml的白细胞介素(IL)-4诱导小鼠骨髓细胞分化为DCs,将其分为空白对照组、胰岛素B9-23多肽刺激组、脂多糖(LPS)刺激组;②流式细胞术检测DCs表面CD11c、CD80、CD86、CD40、MHC-Ⅱ类分子的表达;③CCK-8检测DCs刺激同种异体淋巴细胞增殖的能力;④ELISA检测DCs培养上清中IL-12及干扰素-γ(IFN-γ)的水平。结果:流式细胞术检测各组DCs,其CD11c的表达均在60%以上,与空白对照组相比,胰岛素B9-23多肽刺激组表达低水平的CD40以及中等水平的CD80、CD86、MHC-Ⅱ类分子,LPS刺激组表达较高水平的CD80、CD86、CD40、MHC-Ⅱ类分子,同时,胰岛素B9-23多肽组DCs刺激同种异体淋巴细胞增殖的能力显著增强(P<0.05),分泌较高水平的IL-12,但分泌较低水平的IFN-γ。结论:胰岛素B9-23多肽能够刺激产生半成熟的DCs,对进一步探讨应用胰岛素B9-23多肽负载DCs免疫NOD鼠诱导免疫耐受预防自身免疫性糖尿病的发生具有重要的意义。Objective：To observe the phenotype and functional changes of murine bone marrow derived dendritic cells loaded the insulin B9-23 peptide,and investigate its impact on the immune state of DCs.Methods：①Murine bone marrow cells were induced to differentiate into DCs by 10 ng/ml GM-CSF and 10 ng/ml IL-4,and then devided into blank control group,insulin B9-23 peptide stimu-lated group and LPS stimulated group.②CD40,CD80,CD86,CD11c and MHC-Ⅱexpressions were detected using flow cytometry.③The stimulating allogenic lymphocytes proliferation abilities of these DCs were checked by CCK-8.④ELISA was used to measure IL-12 and IFN-γ concentrations of the DCs culture supernatants.Results：CD11c expression was over 60% in all three groups;compared with the blank control group,the insulin B9-23 peptide group expressed low level CD40 and intermediate level CD80,CD86,MHC-Ⅱ,LPS group DCs expressed CD40,CD80,CD86 and MHC-Ⅱat high level.Meanwhile,DCs in insulin B9-23 peptide group showed increased a bility to stimulate allogenic T cells proliferation,and the IL-12 consentration was significant higher while the IFN-γ concentration was a little lower than the blank control group.Conclusion：Stimulation by lnsulin B9-23 peptide generates semi-mature DCs.It is very meaningful for further studies,in which DCs loaded with insulin B9-23 peptide immune NOD mice to induce immunotolerance to prvent autoimmune diabetes.

关 键 词：树突状细胞 胰岛素B9-23多肽 自身免疫性糖尿病 一级预防 

分 类 号：Q256[生物学—细胞生物学]