草分支杆菌F.U.36对人脐血树突状细胞功能影响的研究

发布时间：2018-03-27 14:29

本文选题：草分支杆菌　切入点：树突状细胞　出处：《青岛大学》2007年硕士论文

【摘要】： 目的探讨草分支杆菌F.U.36(U)对人脐血来源树突状细胞(DC)体外扩增的影响。方法采用体外细胞培养技术，对照组以RPMI-1640培养液诱导脐血单个核细胞(CB-MNC)生成DC，实验各组分别用加有U、重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)+重组人肿瘤坏死因子α(rhTNF-a)+重组人白细胞介素-4(rhIL-4)(GTI)及GTI+U(GTIU)的RPMI-1640培养液诱导CB-MNC生成DC。光镜下观察DC形态。于培养第10天，计数各组中DC，用流式细胞仪检测各组细胞免疫表型，并将细胞涂片行瑞氏-姬姆萨染液染色，，油镜下观察并摄片。结果各实验组均得到一定数量的典型DC；U组CD1a~+、HLA-DR~+细胞比例高于对照组；GTIU组HLA-DR~+细胞比例升高最明显，高于GTI组。结论草分支杆菌F.U.36不仅能促进脐血DC体外扩增，还能协同rhGM-CSF、rhTNF-a、rhIL-4促进DC成熟。目的探讨草分支杆菌F.U.36对人脐血来源树突状细胞(DC)介导的抗白血病效应的影响。方法应用Ficoll-Hypaque法分离人脐血单个核细胞(CB-MNCs)，对照组以重组人粒细胞-巨噬细胞集落刺激因子+重组人白细胞介素-4+重组人肿瘤坏死因子α诱导培养DC，并于DC培养的第4d、第9d加入HL-60细胞。实验组与对照组的细胞因子、HL-60细胞相同，于培养第7d，分别加入不同浓度的草分支杆菌F.U.36混悬注射液，培养不同的时间。光镜下观察DC形态。收获培养第12天的DC，应用流式细胞仪检测DC免疫表型后，将DC与急粒缓解期患儿外周血淋巴细胞混合，诱导抗原特异性细胞毒T淋巴细胞，MTT法检测其杀伤活性。结果草分支杆菌F.U.36浓度与作用时间对CD1a~+细胞比例均无影响。实验组的HLA-DR~+细胞比例及杀伤活性均高于对照组。在一定范围内，随着浓度的增大、作用时间的延长，HLA-DR~+细胞比例增大，杀伤活性亦增强。结论草分支杆菌F.U.36能够促进DC的成熟、增强DC抗原提呈能力，从而增强人脐血来源树突状细胞介导的抗白血病免疫效应。
[Abstract]:Objective to investigate the effect of F. U. 36 UF on the expansion of dendritic cells derived from human umbilical cord blood (UCB) in vitro. Methods Cell culture in vitro was used. In the control group, cord blood mononuclear cells (CB-MNC-) were induced to produce DCs by RPMI-1640 culture medium. The experimental groups were treated with recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF), recombinant human tumor necrosis factor 伪 (rhTNF-a) and recombinant human interleukin-4- (rhIL-4) and GTI, respectively. DC was induced by RPMI-1640 medium. DC morphology was observed under light microscope. After 10 days of culture, DC morphology was observed. The immunophenotypes of each group were detected by flow cytometry, and the cell smear was stained with Riesh-Giemsa staining, observed and photographed under oil microscope. Results the percentage of HLA-DR~ cells in the typical DCU group was higher than that in the control group and higher than that in the GTI group. Conclusion Mycobacterium F.U.36 can not only promote the expansion of umbilical cord blood DC in vitro, but also promote DC maturation in combination with rhGM-CSF and rhTNF-aIL-4. Objective to investigate the effect of mycobacterium F.U.36 on the anti-leukemia effect mediated by dendritic cells derived from human umbilical cord blood. Methods Human umbilical cord blood mononuclear cells (CB-MNCsC) were isolated by Ficoll-Hypaque method. The control group was induced by recombinant human granulocyte-macrophage colony stimulating factor recombinant human interleukin-4 recombinant human tumor necrosis factor 伪 and cultured in DC. HL-60 cells were added on the 4th day and 9th day. The cytokines of the experimental group and the control group were the same as those of the control group. On the 7th day of culture, different concentrations of Mycobacterium graminearum F.U.36 suspension injection were added to culture for different time. The morphology of DC was observed under light microscope. The DC was harvested on the 12th day of culture, and the immunophenotype of DC was detected by flow cytometry. DC was mixed with peripheral blood lymphocytes of children with acute granulocyte remission, and cytotoxic T lymphocytes were induced to detect their cytotoxicity by MTT assay. Results the concentration and time of mycobacterium F.U.36 had no effect on the percentage of CD1a- cells. The percentage and killing activity of HLA-DR~ cells in the experimental group were higher than those in the control group. The percentage of HLA-DR~ cells increased and the cytotoxicity of HLA-DR~ cells increased with the prolongation of time. Conclusion Mycobacterium F.U.36 can promote the maturation of DC, enhance the ability of DC antigen presentation, and enhance the anti-leukemia immunity mediated by dendritic cells derived from human umbilical cord blood.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R392;R378

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