粒细胞-巨噬细胞集落刺激因子质粒对小鼠树突状细胞前体的体内诱导研究  

Study of the inducing effects of GM-CSF plasmid on pre-DC from murine bone marrow in vivo

作　　者：胡微微[1] 臧国庆[1] 余永胜[1] 汤正好[1] 

机构地区：[1]上海市交通大学附属第六人民医院感染科,200233

出　　处：《肝脏》2006年第6期388-390,共3页Chinese Hepatology

基　　金：上海市自然科学基金资助项目(03ZR14066)

摘　　要：目的观察粒细胞-巨噬细胞集落刺激因子(GM-CSF)表达质粒体内转染对小鼠髓源性树突状细胞(DC)前体的诱导作用,为探索乙型肝炎新的免疫治疗提供研究基础。方法30只BALB/c小鼠随机分为对照组、100μg剂量组和200μg剂量组,注射1周,分离骨骼肌,抽提RNA,利用RT-PCR观察反转录水平;分离小鼠骨髓细胞,并以重组小鼠粒细胞/巨噬细胞集落刺激因子(rmGM-CSF)和重组小鼠白细胞介素-4(rmIL-4)诱导培养DC,倒置显微镜下观察其形态,并做扫描电镜检测。ELISA法检测小鼠血清中的GM-CSF表达量,流式细胞仪检测其表面分子(CD80,CD86,MHC-Ⅱ,CD11c)表达情况。结果体内转染后,ELISA检测血清中GM-CSF表达量,与对照组比较,100μg组和200μg组表达量均有明显升高(P<0.01);流式细胞仪检测显示100μg组和200μg组CD11c和MHC-Ⅱ类分子表达较对照组明显升高(P<0.01),CD80和CD86有所升高(P<0.05)。结论GM-CSF质粒体内注射可以明显刺激诱导小鼠髓源性DC前体的增殖,增强DC的抗原提呈作用。Objective To investigate the inducing effects in vivo of the transfection of GM-CSF plasmid on murine marrow pre- DC in order to explore some basis for new immune treatment of chronic hepatitis B. Methods 30 BALB/c mice were randomly divided into three groups：normal control group, 100 μg group and 200 μg GM-CSF plasmid group. One week after infection,mice were killed, RNA extracted from skeletal and muscle, level of inverse transcription investigated by RT-PCR. Mice bone marrow cells were isolated and, induced and cultured with rmGM-CSF and rmIL-4. Inverted microscope and Scanning electron microscope were used to investigate the morphous of DC cells. The phenotypes of DC was detected by flow cytometry and the expression of the GM-CSF protein in serum was tested by ELISA. Results The expression of the GM-CSF protein in serum in 100 μg group and 200 μg GM-CSF plasmid group were both increased significantly compared with the normal control group. The degree of CD1 lc and MHC-Ⅱ expressions in 100 μg and 200 μg groups on flow cytometry were elevated significantly compared with that in control group, and the degree of CD80 and CD86 was increased. Conclusion The transfection of the GM-CSF plasmid in vivo could stimulate the proliferation of the pre DC of bone marrow, and enhance antigen presentation of DC.

关 键 词：粒细胞-巨噬细胞集落刺激因子 树突状细胞 体内诱导 

分 类 号：R512.62[医药卫生—内科学] R395[医药卫生—临床医学]