兰州大学机构库 >学院待认领
香菇多糖对急性髓系白血病患者树突状细胞的成熟和功能影响的体内外研究
Alternative TitleThe maturation and function study of lentinan on dendritic cells of acute myeloid leukemia patients in vitro and in vivo
王玉虹
Thesis Advisor张连生
2010-05-25
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword香菇多糖 树突状细胞 急性髓系白血病
Abstract目的:通过体内外试验研究香菇多糖(lentinan,LNT)对急性髓系白血病(AML)患者树突状细胞(DC)的促成熟及功能调节的机制,进一步阐明香菇多糖的免疫学活性,并探索白血病免疫治疗的新途径。 方法:体外试验用Ficoll密度梯度离心法分离急性髓系白血病完全缓解期(AML-CR)患者的骨髓单个核细胞(BMNC);用含重组人粒-巨噬细胞集落刺激因子(rhGM-CSF)1000U/ml和重组人白细胞介素4(rhIL-4)500U/ml的RPMI-1640完全培养基在37℃ 5%CO2孵箱中培养至第7天。阳性对照组加常规量的脂多糖(LPS),实验组分别用LNT1ug、3ug、5ug刺激DC,阴性对照组加相同体积的培养液,培养48小时后;经形态学观察(倒置显微镜、Wright-Giemsa染色),免疫学(CD80、CD83、CD86、CD1a、HLA-DR)的鉴定,ELISA双抗体夹心法检测DCs上清液白细胞介素12(IL-12)的水平。体内试验用免疫磁珠法(MACS)分离AML-CR患者经LNT治疗前、后的外周血单个核细胞(PBMNC)的DCs;ELISA双抗体夹心法检测DCs上清液IL-12的水平。 结果:体外试验,高剂量组LNT作用的DC呈典型形态;能够提高DC表面分子CD80、CD83、CD86、CD1a及HLA-DR的表达(P<0.05);DC分泌IL-12较阴性对照组显著增高(P<0.05),且呈LNT浓度依赖性。体内试验,经LNT治疗后分离纯化的DC分泌IL-12的量与治疗前比较有明显差异(P<0.05),与体外试验相一致。 结论:LNT在体内、外均能促进AML患者DC的成熟和功能的增强而间接发挥抗白血病作用。
Other AbstractObjective To explort the regulatory effects of lentinan (LNT) on maturation and function of dendritic cells (DC) of acute myeloid leukemia patients in vitro and in vivo, further clarify the immunological activity of lentinan, and to explore new ways of leukemia immunotherapy. Method Bone marrow mononuclear cells (BMNCs) from acute myeloid leukemia complete remission (AML-CR) patients were isolated with Ficoll density gradient centrifugation method,and were cultured in RPMI-1640 medium containing recombinant human granulocyte colony stimulating factor (GM-CSF)/recombinant human interleukin 4 (IL-4) placed in 5%CO2 air at 37℃ incubator. After culture for 7 days,positive control group was affected by a conventional dose of lipopolysaccharide (LPS), experimental groups were stimulated by LNT at 1ug,3ug and 5ug dose,negative control group was treated by RPMI-1640 medium of an equal volume.After 48 hours of stimulation, Morphology of DCs was observed under inverted microscope and Wright-Giemsa staining respectively. Phenotypes on the cells were identified by flow cytometer.Cell populations were phenotyped with the following FITC- or PE-conjugated MoAbs:CD80, CD83, CD86, CD1α, HLA-DR. IL-12 production was detected by ELISA. Dendritic cells in complete remission acute myeloid leukemia patients were isolated from human peripheral blood mononuclear cells (PBMNC) by magnetic cell sorting(MACS),we analyzed the concentration of IL-12 secreted by DCs using ELISA. Results In vitro effect of high doses of LNT showed typical DC morphology,could increase the expression of CD80,CD83,CD86,CD1a and HLA-DR molecules on DC surface(P<0.05),LNT-treated DCs secreted higher level of IL-12 than negative control group(P<0.05),and was concentration-dependent Lentinan.LNT-treated DCs secreted significantly higher level of IL-12 than untreated DCs(P<0.05),consistent with in vivo and in vitro. Conclusion LNT could promote functional and phenotypic maturation of dendritic cells of acute myeloid leukemia patients in vivo and in vitro,LNT might be one of its anti-leukemia mechanism.
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/222511
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
王玉虹. 香菇多糖对急性髓系白血病患者树突状细胞的成熟和功能影响的体内外研究[D]. 兰州. 兰州大学,2010.
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