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树突状细胞负载肺癌细胞抗原的方法研究 被引量:1

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摘要 目的:建立自体热休克凋亡肺癌细胞抗原制备、树突状细胞(DC)体外诱导、以及抗原负载方法,为制备DC肿瘤疫苗提供技术基础。方法:采用酶消化法从手术切除的肺癌新鲜组织获得单细胞悬液,热休克后用白桦脂酸诱导其凋亡制备成细胞抗原;采用血细胞分离机采集外周血单个核细胞(PBMC),贴壁法获得单核细胞,经GM-CSF与IL-4体外诱导成未成熟树突状细胞(imDC);负载细胞抗原后制备成DC肿瘤疫苗,并对DC疫苗的形态、数量及表型特征进行分析。结果:(1)肿瘤细胞抗原得率:(13.68±1.30)×106/g组织;平均凋亡率:(93.79±2.31)%;(2)imDC平均得率为(9.37±0.83)×106/(118.77±12.56)×106个PBMC,活率>95%;imDC表型分析:CD11c+CD14-、CD11c+HLA-DR+、CD11c+CD80+、CD11c+CD83+、CD11c+CD86+表达率分别为(87.45±3.42)%、(87.16±1.08)%、(2.80±1.52)%、(5.64±1.56)%、(5.11±1.09)%;(3)DC疫苗平均得率为(5.75±0.69)×106/(9.37±0.83)×106个imDC,活率>95%,DC疫苗表型:CD11c+CD14-、CD11c+HLA-DR+、CD11c+CD80+、CD11c+CD83+、CD11c+CD86+表达率分别为(92.73±1.21)%、(89.35±2.31)%、(86.80±1.23)%、(69.53±7.21)%、(94.92±1.48)%。结论:该方法稳定、安全、可靠,可制备出具有成熟DC表型的肿瘤疫苗。
出处 《细胞与分子免疫学杂志》 CAS CSCD 北大核心 2008年第11期1113-1115,共3页 Chinese Journal of Cellular and Molecular Immunology
基金 南京市科技人才创新创业项目(20060288) 江苏省常州市卫生局重大科研项目(ZD200604)
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  • 1Jefford M, Maraskovsky E, Cebon J, et al. The use of dendritic cells in cancer therapy[J]. Lancet Oncol, 2001, 2(6) : 343 -353.
  • 2Gliboa E. DC-based cancer vaccines[J]. J Clin Invest, 2007, 117 (5) : 1195 -1203.
  • 3Moiseyenko V, Imyanitov E, Danilova A, et al. Cell technologies in immunotherapy of cancer[ J]. Adv Exp Med Biol, 2007, 601 : 387 - 393.
  • 4Vulink A, Radord K J, Melief C, et al. Dendritic cells in cancer immunotherapy[ J]. Adv Cancer Res, 2008, 99: 363- 407.
  • 5Ladhams A, Schmidt C, Sing G, et al. Treatment of non-resectable hepatocellular carcinoma with autologus tumor-pulsed dendritic cells [J]. J Gastroenterol Hepatol, 2002, 17(8) : 889 -896.
  • 6Yu JS, Liu GT, Ying H, et al. Vaccination with tumor lysate-pulsed dendritic cells elicits antigen-specific, cytotoxic T-cells in patients with malignant glioma [ J ]. Cancer Res, 2004, 64 ( 14 ) : 4973 - 4979.
  • 7Shi HZ, Cao TH, Connolly JE, et al. Hyperthermia enhances CTL cross-priming[J]. Immunol, 2006, 176(4): 2134-2141.
  • 8孟淑芳,林林,李修兰,冯建平,王佑春,李德富.软琼脂克隆法与裸鼠体内接种法检测细胞致瘤性的比较[J].中国生物制品学杂志,2006,19(5):516-519. 被引量:22
  • 9Paczesny S, Shi HZ, Saito H, et al. Measuring melanoma-specific cytotoxic T lymphocytes elicited by dendritic cell vaccines with a tumor inhibition assay in vitro [ J ]. J Immunother , 2005, 28 ( 2 ) : 148 - 157.
  • 10Laberu MS, Roters B, Pers B, et al. Generation of tumor immunity by bone marrow-derived dendritic cells correlates with dendritic cell maturation stage[ J]. J Immunol, 1999, 162 ( 1 ) : 168 - 175.

二级参考文献6

  • 1WHO Technical Report Series,No.878,1998,Annex 1:Requirements for the used of animal cells as in vitro substrated for the production of biologicals.
  • 2Davis JM.Basic cell culture.Oxford University Press,2002.
  • 3FDA.Points to consider in the characterization of cell lines used to produce biologicals.1993.
  • 4Hirose K,Hakozaki N,Nyunova Y,et al.Chemokine gene transfection into tumout cells reduced tumorigenicity in nude mice in association with neurtrophilic infiltration.Br J Cancer,1995,72 (3):708-714.
  • 5Vincent-Falquet JC,Peyron L,Souvras M,et al.Qualification of working cell banks for the vero cell line to produce licensed human vaccines.Developments in Biological Standardization,1989,70:153-156.
  • 6Petricciani JC,Levenbook IS,Wierenga DE,et al.Early passage primate cell immotality is independent of tumorigenicity.In Vitro Cell Dev Biol,1987,23(7):523-526.

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