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作 者:庄泳[1] 李栋[2] 付金秋[1] 时庆[2] 鞠秀丽[1]
机构地区:[1]山东大学齐鲁医院儿童医疗中心,山东济南250012 [2]山东大学齐鲁医院低温医学研究室,山东济南250012
出 处:《山东大学学报(医学版)》2014年第11期60-64,共5页Journal of Shandong University:Health Sciences
基 金:山东省自然科学基金(ZR2011HM007);山东省科技攻关计划(2013GSF11812)
摘 要:目的体外诱导培养正常儿童与B系急性淋巴细胞白血病(B-ALL)儿童的树突状细胞(DC),比较二者的生物学特性。方法分离10例B-ALL初诊患儿(ALL组)和10例正常儿童(对照组)的外周血单个核细胞,以rh GM-CSF(20 ng/m L)、rh IL-4(10 ng/m L)及TNF-α(10 ng/m L)联合培养8 d,显微镜下观察细胞形态,流式细胞仪检测细胞免疫表型(CD11c、CD80、CD83、CD86),ELISA检测培养上清中IL-12的浓度,混合淋巴细胞反应(MLR)检测抗原递呈功能,电化学法检测上清中葡萄糖浓度。结果 ALL组细胞未表现出DC的典型形态,CD11c、CD80、CD83、CD86表达均较对照组细胞低(P<0.05),分泌IL-12的能力弱于对照组细胞(P<0.05),刺激T淋巴细胞增殖的能力弱于对照组细胞,ALL组细胞培养上清中的葡萄糖浓度高于对照组(P<0.05)。结论B-ALL来源的DC成熟度异常,功能减弱,葡萄糖代谢异常可能是其成熟异常的原因之一。Objective To induce and culture dendritic cells (DC)derived from normal children and childhood B line-age acute lymphoblastic leukemia (B-ALL)patients,and compare their biological features in vitro.Methods Periph-eral blood mononuclear cells (PBMCs)were isolated from 10 B-ALL patients (ALL group)and 10 healthy donors (control group).The isolated PBMCs were co-cultured with rhGM-CSF(20 ng/mL),rh IL-4(10 ng/mL)and TNF-α(10 ng/mL)for 8 days.The morphological features were observed by inverted microscope.CD11c,CD80,CD83, CDD86 expressions were assayed by flow cytometry.The concentration of IL-12 was measured by ELISA.The antigen presenting function of the cells were tested by mixed lymphocyte reaction(MLR).Electrochemical measurement was used for the detection of glucose metabolism.Results The typical DC morphological features were not observed from the cells in ALL group.The cells in ALL group expressed lower levels of CD11c,CD83,CD80 and CD86 than the cells in control group(P〈0.05).The cells in ALL group secreted lower level of IL-12 than control group(P〈0.05). The cells in ALL group displayed weaker activity of stimulating T lymphocyte proliferation than control group in MLR test.Glucose concentration in culture medium of ALL group was higher than that of control group (P 〈0.05 ). Conclusion DC derived from B-ALL patients displays a less mature phenotype with compromised functions.Low glu-cose metabolism ability might be a possible reason for the DC defects in ALL.
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