白细胞介素-10诱导的大鼠树突状细胞体外免疫功能的研究

Immune tolerance induced by rat immature dendritic cells treated with interleukin-10 in vitro

目的 研究白细胞介素 10 (IL 10 )诱导的大鼠未成熟树突状细胞 (imDCs)体外诱导免疫耐受的可行性。方法 在经典诱导方案的基础上 ,应用IL 10 ( 10 μg/L)抑制大鼠骨髓来源DCs的成熟 (IL 10组 ,10例 ) ,并设对照组 (IL 4组 ,10例 )。培养期间观察DCs形态 ,检测DCs表型、摄取抗原能力、体外免疫功能及培养上清细胞因子水平。结果 与IL 4组比较 ,IL 10组DCs细胞表面CD80 、CD86及OX6低度表达 ( 2 5 .3 %、42 .4%、3 2 .3 % ) ,吞噬能力较强 ( 81.9) ,刺激同种异体淋巴细胞增殖能力下降 ,该淋巴细胞具有抗原特异性低反应性 ;培养上清中IL 12水平 ( 4 0 6.5pg/L)及初次MLR培养上清IL 2水平 ( 2 45 .4ng/L)均较低 ,差异有非常显著性 (P <0 .0 1)。 结论 IL 10作用的大鼠imDCs具有诱导免疫耐受的应用价值。

Objective To study the feasibility of immune tolerance induced by rat immature dendritic cells (DCs) in vitro treated with interleukin-10.Methods Bone marrow derived DCs progenitors were assigned to two groups: one was cultured with granulocyte macrophage colony stimulating factor (10 μg/L) and interleukin 4 (8 μg/L) for 13 days (IL 4 group),the other with the same cytokines for the first 5 days,then with interleukin 10 (10 μg/L) additionally for the subsequent 8 days (IL 10 group).Cell morphology was monitored under light microscopy and cell surface molecules including OX6,OX62,CD 80 and CD 86 were assayed by flow cytometry.The capacity of DCs uptaking antigens was measured by FITC Dextran endocytosis.T cell proliferation was assayed by 3H thymidin deoxyribose insertion (3.7×10 4 Bq/cell).The concentrations of IL 2 and IL 12 in the supernatant were assayed by ELISA.Results Compared with DCs in IL 4 group,DCs in IL 10 group were characterized as follows: (1) Lower expression levels of CD 80 ,CD 86 and OX6 (25.3%,42.2% and 32.3% respectively); (2) Powerful ability of endocytosis (81.9); (3) Decreased secretion of IL 12 (406.5 pg/L) and IL 2 (245.4 ng/L); (4) Weaker activity of stimulating proliferation of allogeneic T cells; and (5) T cells hyporesponsiveness was donor specific.Conclusion The immature DCs inhibited by IL 10 may have the properties of inducing allogeneic specific immune tolerance in vitro.