本文選題：草分支桿菌　切入點(diǎn)：樹突狀細(xì)胞　出處：《青島大學(xué)》2007年碩士論文

【摘要】： 目的探討草分支桿菌F.U.36(U)對(duì)人臍血來源樹突狀細(xì)胞(DC)體外擴(kuò)增的影響。 方法采用體外細(xì)胞培養(yǎng)技術(shù)，對(duì)照組以RPMI-1640培養(yǎng)液誘導(dǎo)臍血單個(gè)核細(xì)胞(CB-MNC)生成DC，實(shí)驗(yàn)各組分別用加有U、重組人粒細(xì)胞-巨噬細(xì)胞集落刺激因子(rhGM-CSF)+重組人腫瘤壞死因子α(rhTNF-a)+重組人白細(xì)胞介素-4(rhIL-4)(GTI)及GTI+U(GTIU)的RPMI-1640培養(yǎng)液誘導(dǎo)CB-MNC生成DC。光鏡下觀察DC形態(tài)。于培養(yǎng)第10天，計(jì)數(shù)各組中DC，用流式細(xì)胞儀檢測(cè)各組細(xì)胞免疫表型，并將細(xì)胞涂片行瑞氏-姬姆薩染液染色，，油鏡下觀察并攝片。 結(jié)果各實(shí)驗(yàn)組均得到一定數(shù)量的典型DC；U組CD1a~+、HLA-DR~+細(xì)胞比例高于對(duì)照組；GTIU組HLA-DR~+細(xì)胞比例升高最明顯，高于GTI組。 結(jié)論草分支桿菌F.U.36不僅能促進(jìn)臍血DC體外擴(kuò)增，還能協(xié)同rhGM-CSF、rhTNF-a、rhIL-4促進(jìn)DC成熟。 目的探討草分支桿菌F.U.36對(duì)人臍血來源樹突狀細(xì)胞(DC)介導(dǎo)的抗白血病效應(yīng)的影響。 方法應(yīng)用Ficoll-Hypaque法分離人臍血單個(gè)核細(xì)胞(CB-MNCs)，對(duì)照組以重組人粒細(xì)胞-巨噬細(xì)胞集落刺激因子+重組人白細(xì)胞介素-4+重組人腫瘤壞死因子α誘導(dǎo)培養(yǎng)DC，并于DC培養(yǎng)的第4d、第9d加入HL-60細(xì)胞。實(shí)驗(yàn)組與對(duì)照組的細(xì)胞因子、HL-60細(xì)胞相同，于培養(yǎng)第7d，分別加入不同濃度的草分支桿菌F.U.36混懸注射液，培養(yǎng)不同的時(shí)間。光鏡下觀察DC形態(tài)。收獲培養(yǎng)第12天的DC，應(yīng)用流式細(xì)胞儀檢測(cè)DC免疫表型后，將DC與急粒緩解期患兒外周血淋巴細(xì)胞混合，誘導(dǎo)抗原特異性細(xì)胞毒T淋巴細(xì)胞，MTT法檢測(cè)其殺傷活性。 結(jié)果草分支桿菌F.U.36濃度與作用時(shí)間對(duì)CD1a~+細(xì)胞比例均無影響。實(shí)驗(yàn)組的HLA-DR~+細(xì)胞比例及殺傷活性均高于對(duì)照組。在一定范圍內(nèi)，隨著濃度的增大、作用時(shí)間的延長(zhǎng)，HLA-DR~+細(xì)胞比例增大，殺傷活性亦增強(qiáng)。 結(jié)論草分支桿菌F.U.36能夠促進(jìn)DC的成熟、增強(qiáng)DC抗原提呈能力，從而增強(qiáng)人臍血來源樹突狀細(xì)胞介導(dǎo)的抗白血病免疫效應(yīng)。
[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 偽 (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 偽 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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R392;R378

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