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.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
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)
关键词
树突状细胞
粒巨噬细胞集落刺激因子
免疫耐受
Dendritic cells
Granulocyte-Macrophage Colony-Stimulating Factor
Immune tolerance