低剂量粒巨噬细胞集落刺激因子培养小鼠骨髓源性未成熟树突状细胞的实验研究  

Generation of immature mouse myeloid dendritic cells with low-dose granulocyte-macrophage colony-stimulating factor

作　　者：冯剑锷[1] 孙宗全[1] 史嘉玮[1] 高开柱[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院心血管研究所心外科,武汉430022

出　　处：《中华实验外科杂志》2006年第2期221-223,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目(30471715)

摘　　要：目的建立小鼠骨髓源性未成熟树突状细胞(DC)的培养方法,并初步观察其生物学特性。方法分别用常规剂量和低剂量粒巨噬细胞集落刺激因子(GM-CSF)培养小鼠骨髓源性DC,对其进行形态学观察和细胞表型检测,检测其在体外刺激同种异体T细胞增殖的能力,同时比较不同剂量GM-CSF下培养细胞内IL-10和TGF-βmRNA的表达水平。结果常规剂量GM-CSF培养出的DC为成熟DC和未成熟DC的混合体,经脂多糖(LPS)刺激后迅速分化成熟,表型为CD11c+、CD25±、CD80+、CD86+、MHCⅡhi,在体外可强烈刺激同种异体T细胞分化增殖;而低剂量GM-CSF培养出的DC为较单纯的未成熟DC,LPS不能刺激其分化成熟,表型为CD11c+、CD25-、CD80-、CD86-、MHCⅡlow,不能有效活化刺激同种异体T细胞,其IL-10mRNA表达水平明显高于其他细胞。结论用低剂量GM-CSF可培养出表型和功能均未成熟的小鼠骨髓源性DC,其自身高水平分泌IL-10可能是其成熟障碍的原因。Objective To establish a simple method to generate immature dendritic cells with low-dose granulocyte-macrophage colony-stimulating factor （GM-CSF） from mouse bone marrow cells and observe their biological features. Methods Mouse DCs were generated with standard doses or low doses of GM-CSF from bone marrow cells separately. The phenotype and functional properties of these DCs were compared through FACS analysis and MLR, and the expression levels of IL-10 and TGF-β mRNA in cultured cells were detected using RT-PCR. Results DCs generated with standard doses of GM- CSF were a mixture of mature DCs and immature DCs could be induced to maturation by lipopolysaccharide （LPS）, and they were strong stimulators of allogeneic T cell responses, whereas DCs generated with low doses of GM-CSF were phetotypically immature DCS （CD11c^＋、CD25^-、CD80^-、CD86^-、MHC Ⅱ^low）, and induced allogeneic T cell unresponsiveness, and the expression level of IL-10 mRNA in these DCs was much higher than the other cells. Conclusion DCs generated from mouse bone marrow progenitors in low doses of GM-CSF were phenotypically and functionally immature, and their resistance to maturation stimulation may be due to high level of self-secretion of IL-10.

关 键 词：树突状细胞 粒巨噬细胞集落刺激因子 免疫耐受 

分 类 号：R392.12[医药卫生—免疫学]