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目的 研究IL-24基因修饰的树突状细胞(DC)与同源细胞因子诱导的杀伤细胞(cyto-kine-induced killer,CIK)共培养后对A549肺癌细胞的杀伤作用及其机制.方法 从健康人外周血单个核细胞中常规诱导DC、CIK细胞,同时抽提重组腺病毒质粒pAdEasy-1-pTrack-CMV-IL-24,Pac I酶切线性化后脂质体转染QBl-293A细胞,收获Ad-IL-24重组病毒.将IL-24基因通过重组病毒导入已负载肿瘤抗原的DC,获得细胞称为DC-IL-24.RT-PCR和ELISA法检测DC中IL-24基因的表达,FCM和ELISA法检测DC表型及分泌细胞因子能力的变化,将DC和CIK细胞混合培养,溶血试验检测CIK细胞产生穿孔素的能力,FCM法检测共培养的DC-CIK细胞对A549肺癌细胞细胞毒活性的变化.结果 获得了高滴度的重组病毒Ad-IL-24并成功将IL-24基因导入DC,在倒置荧光显微镜下可观察到荧光,IL-24可上调Dc表而CD80、CD83、HLA-DR、CD40、CXCR4分子的表达,转染后Dc分泌IL-12、TNF-α和IL-24的能力显著增强,DC-IL-24更能促进CIK细胞产生穿孔素,与同源CIK细胞共培养后对A549肺癌细胞的细胞毒活件明显增强.结论 IL-24基因修饰的DC能增强自体CIK细胞产生特异性抗肿瘤免疫,其机制与IL-24能促进DC表型成熟,分泌Th1型细胞因子,维持DC活化状态,进而促进CIK细胞活化密切相关. 相似文献
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随着医疗技术水平的不断提高,浓缩血小板的临床使用量不断增加,为了解决血小板的应急供应,一些采供血单位开展了血小板的冰冻保存工作。为了观察冰冻保存对血小板活化的影响,笔者检测了冰冻保存前后的血小板表面CD62分子表达,现报告如下。 相似文献
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《全血及成分血质量要求》GB18469-2012中,对红细胞制品的容量有具体规定,均要求容量(ml)是标示量±10%[1]。但是,红细胞制品是以U(单位)作为标示量,而由称重法得到的容量又是以ml为单位。因此,在实际工作中,国家标准作为红细胞制品的容量判断的依据,在操作上欠可行。 相似文献
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目的:研究IL-24基因修饰的CIK细胞与同源树突状细胞共培养后对白血病细胞的杀伤作用及其机制.方法:从健康人外周血单个核细胞中常规诱导DC和CIK 细胞,电穿孔法将IL-24基因导入CIK细胞中(获得细胞为CIK-IL24),RT-PCR 和ELISA法检测CIK细胞中IL-24基因的表达,FCM和ELISA法检测转基因前后CIK表型及分泌细胞因子能力的变化,将CIK 细胞和同源DC共培养,FCM法检测共培养的DC-CIK细胞对HL-60细胞细胞毒活性的变化.结果:通过电穿孔法成功将IL-24基因导入CIK细胞,与对照组相比,转IL-24基因后CIK细胞中CD3~+、CD3~+CD56~+细胞的比例无明显改变,CD4~+CD25~+细胞比例显著下降.IL-24可上调CD3+CD56+细胞表面粘附分子CD54、CXCR4的表达,转染IL-24基因后CIK分泌TNF-α和IFN-γ的能力显著增强,与DC共同作用HL-60细胞时转染IL-24基因后的CIK细胞细胞毒活性明显增强.结论:通过IL-24基因修饰,明显增强了CIK细胞对HL-60细胞的杀伤能力,其机制与IL-24促进CIK分泌TNF-α、IFN-γ,上调CIK细胞表面粘附分子的表达,减少CD4~+CD25~+调节性T细胞比例等密切相关. 相似文献
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Objective To study the antitumor effect and mechanism of co-cultured cytokine-induced killer(CIK) cells and autologous DC modified with IL-24 gene on A549 cells in vitro. Methods DC and CIK cells were prepared routinely from human peripheral blood mononuclear cells(PBMC). Recombinant adenovirus vector pAdEasy-1-pTrack-CMV-IL-24 was extracted from DH5α, it was lineared with Pac I and transfected into A293 cells, and then the IL-24 recombined adenovirus(Ad-IL-24) was obtained. Ad-IL-24 was used to infect DC. The cells obtained were named DC-IL-24. RT-PCR and ELISA were used to evaluate the expression of IL-24 gene in transfected DC. The phenotypes change of DC were identified by flow cytometry analysis, the concen-tration of IL-12 and TNF-α in supernatant of DC were determined by EIJSA. The ability of CIK producing per-forin was measured by homolysis method. FCM was used to determine the cytotoxicity of cocultured CIK cells and autologous DC modified with IL-24 gene to A549 cells. Results We obtained the high titre of Ad-IL-24.IL-24 gene was transfered into DC successfully via Ad-IL-24. The green fluorescence was observed on DC by fluorescence microscope. The expression rate of CD80, CD83, HI.A-DR, CD40, CXCR4 on DC-IL-24 was sig-nificantly increased compared with that of the control group. DC-IL-24 produced markedly higher levels of IL-12 and TNF-α as compared with DC. DC-IL-24 can enhance the ability of CIK cells producing perforin. On com-parison with non-transfected DC co-cultured with CIK cells, transfected DC co-cultured with CIK cells had a sig-nificantly higher lytic activity against A549 cells. Conclusion IL-24 gene modification can enhance the anti-tu-moral immunity of DC. The mechanism of which might be related to the increased secretion of IL-12 and TNF-α, up-regulation expression of co-stimulatory molecules and MHC Ⅱ class molecules on DC, promoting the acti-vation and maturation of DC, and then enhancing CIK cells to generate specific anti-tumoral immunity. 相似文献
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不同方法制备的浓缩血小板表面CD62分子的表达 总被引:4,自引:1,他引:4
目的 了解不同制备方法对浓缩血小板表面CD6 2表达的影响。方法 采用藻红素标记的抗CD6 2单抗 ,经流式细胞仪分析血小板表面CD6 2表达情况。结果 (6 .5 5± 3.5 2 ) %的机采血小板表达CD6 2 ,(9.2 6± 5 .4 0 ) %的白膜回浆法 (BC法 )血小板表达CD6 2 ,高达 (5 0 .30± 2 0 .4 2 ) %的富血小板血浆法 (PRP)血小板表达CD6 2。结论 相对于PRP法 ,机采法和BC法更适合于浓缩血小板的制备。 相似文献
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目的了解2%(v/v)DM SO结合第二信使效应剂冰冻保存浓缩血小板的可行性。方法通过在含2%(v/v)DM SO的浓缩血小板中加入不同浓度的第二信使效应剂,经-80℃冰冻保存1 w,然后分别测定PLT、M PV、L eakage of LDH及血小板表面CD 62的表达。结果硝普钠(SNP)达25μm o l/L时能显著降低CD 62分子的表达(P<0.05)。结论SNP对血小板在2%(v/v)DM SO条件下-80℃冰冻保存的效果有所改善,但氨氯吡脒(A 7410)的效果不明显。 相似文献