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脂质体RNA负载慢性粒细胞白血病来源树突状细胞的性能比较

Anti-Chronic Myelocytic Leukemia Induced by Dentritic Cells Transfected with RNA-lipofectamin
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摘要 目的研究人慢性粒细胞白血病(CML)总RNA经过脂质体负载后体外转染自体慢性粒细胞白血病-树突状细胞(CML-DC),并诱导特异性细胞毒T淋巴细胞(CTL)免疫反应。方法采用Trizol法提取CML-BMMNC的总RNA;反复冻融法制备CML-BMMNC抗原。于CML-DC培养第5天,加入脂质体转染的CML总RNA、CML总RNA、CML肿瘤冻融抗原及不加抗原。倒置显微镜观察DC形态学变化;流式细胞仪器检测免疫表型变化;染色体G显带技术检测其染色体核型及逆转录聚合酶链反应(RT-PCR)检测Bcr-abl融合基因的表达;用MTT法检测CTL作用。结果CML BMMNC诱导成CML-DC,CD1α、CD83表型表达较诱导前明显上调,形态学有典型的DC形态,且均存在Bcr-abl基因带和Ph1染色体,提示CML-DC为白血病源性。总RNA经脂质体转染CML-DC、CML肿瘤冻融抗原负载CML-DC、总RNA不经脂质体转染CML-DC、未负载抗原CML-DC分别致敏的CTL及IL-2培养的T淋巴细胞在效:靶比为20∶1时的杀伤效率依次降低。结论CML BMMNC来源的DC既具有CML白血病源性,又具有DC细胞的特性,能诱导特异性CTL杀伤作用;总RNA经脂质体转染的CML-DC诱导的CTL杀伤性对CML细胞的杀伤作用最强。 Objective To investigate the specific cytotoxic T lymphocyte(CTL) irnmunoreaction of choronic myeloeytie leukemia-dendritic cells(CMLq3C) transfected with CIVIL total RNA in vitro, and to provide a theoretic basis of CML vaccine in clinical study. Metheds Trizol method was used to extract CML-BMMNCs total RNA, and CML-BMMNCs were applied to prepare tumor frozen-thawed antigen. At the 5th day of culture, CML-DCs were divided into four groups: CML-DCs pulsed with total RNA-lipofectamin; CML-DCs pulsed with total RNA; CML-DCs pulsed with tumor frozen-thawed antigen and CML-DCs pulsed with nothing. All of them could induce T cells into specific CTL. T cells cultured with IL-2 served as normal control group. The morphological features of DCs were observed under an inverted micmscope. The CD1a and CD83 were analyzed by flow eytometry. Karyotypes of CML-DC were tested by G-banding technique The Bcr-abl expression in CML-DCs was detected by reversed transcription polymerase chain reaction (RT-PCR). MTT assay was used to test the capacity of CML-DCs for mixed leukocyte reaction(MLR). Results CML-BMMNCs could be induced into CML-DCs. The expression levels of CD, a and CD83 in CML-DCs were increased obviously as com- pared with those in uncultured CML-BMMNCs. Bcr-abl and Ph chromosome all exsisted in cultured CML-DCs and uncultured CML-BMMNCs. It was suggested that CML-IX2s were from CML cells. The cytotoxie rate of CML-DCs pulsed with total RNA-lipofectamin, CML-DCs pulsed with total RNA, CML-DCs pulsed with tumor frozen-thawed antigen, CML-DCs pulsed with nothing, and T cells cultured with IL-2 was decreased in turn. Conclusion CML-BMMNCs-derived DCs could induce the CTL to get specific anti-tumor activity in vitro. The eytotoxieity of CTL induced by CML-DCs pulsed with total RNA-lipofectamin was the most significant.
出处 《肿瘤防治研究》 CAS CSCD 北大核心 2010年第1期47-51,共5页 Cancer Research on Prevention and Treatment
基金 江西教育厅科学技术研究资助项目(赣教技字2006-87)
关键词 树突状细胞 慢性粒细胞白血病 总RNA Dendritic cell Chronic myelocytic leukemia Total RNA
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  • 1Mcmichael AJ. The original sin of killer T cells[J]. Nature, 1998,394(6692) :421-422.
  • 2Girolomoni F, Castagnoli PR. Dendritic cells hold promise for immunotherapy[M]. Mmunol Today, 1997,18 (3) : 102- 104.
  • 3王俊祥,王金铠,孟建波.慢性髓性白血病来源的树突状细胞诱导的CTL体外作用研究[J].肿瘤防治研究,2004,31(12):739-741. 被引量:1
  • 4许思娟,张连生,吴重阳,柴晔,宋飞雪,岳玲玲,刘瑛.抑制吲哚胺2,3-双加氧酶活性促进慢性粒细胞白血病源树突状细胞的功能[J].中国肿瘤生物治疗杂志,2009,16(2):170-174. 被引量:3
  • 5Saenz Badillos J, Amin SP, Granstein RD. RNA as a tumor vaccine: a review of the literature[J]. Exp Dermatol,2001.10 (3):143 154.
  • 6Kariko K, Kuo A, Barnathan ES, ctal. Phosphate enhanced transfection of cationic lipid complexed mRNA and plasmid DNA[J]. Biochim Biophys Acta, 1998, 369(2): 320-334.
  • 7Mitchell DA. Nair SK. RNA transfected dendriticcells in cancer immunotherapy[J]. J Clin Invest, 2000,1 06(9). 1 065- 1069.
  • 8Syme R, Bryan T, Duggan P, et al. Priming with Dendritic Cells Can Generate Strong Cytotoxic T Cell Responses to Chronic Myelogenous Leukemia Cells In vitro[J]. Stem cells and development, 2004,13(2) :211- 221.
  • 9Heiser A, Maurice MA, Yancey DR, et al. Human dendritic cells transfected with renal tumor RNA stimulate polyclonal T- cell responses against antigens expressed by primary and metastatic tumor[J]. Cancer Res, 2001, 61 (8) :3388 -3393.
  • 10Boczkowski D, Nair SK, Nam JH, et al. Induction of tumor immunity and cytotoxic T lymphocyte responses using dendrit ic ceils transfected with messenger RNA amplifid from tumor cells[J]. Cancer Res, 2000, 60(4) : 1028 -1034.

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