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首页> 外文期刊>Biochimica et biophysica acta. Molecular basis of disease: BBA >Human tumour and dendritic cell hybrids generated by electrofusion: potential for cancer vaccines
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Human tumour and dendritic cell hybrids generated by electrofusion: potential for cancer vaccines

机译：电融合产生的人类肿瘤和树突状细胞杂种：癌症疫苗的潜力

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Hybrid cells created by fusion of antigen presenting and tumour cells have been shown to induce potent protective and curative anti-tumour immunity in rodent cancer models. The application of hybrid cell vaccines for human tumour therapy and the timely intervention in disease control are limited by the requirement to derive sufficient autologous cells to preserve homologous tumour antigen presentation. In this study, the efficiency of various methods of electrofusion in generating hybrid human cells have been investigated with a variety of human haemopoiletic, breast and prostate cell lines. Cell fusion using an electrical pulse is enhanced by a variety of stimuli to align cells electrically or bring cells into contact. Centrifugation of cells after an exponential pulse from a Gene Pulser electroporation apparatus provided the highest yield of mixed cell hybrids by FACS analysis. An extensive fusogenic condition generated in human cells after an electrical pulse contradicts the presumption that prior cell contact is necessary for cell fusion. Alignment of cells in a concurrent direct current charge and osmotic expansion of cells in polyethylene glycol also generated high levels of cell fusion. Waxing of one electrode of the electroporation cuvette served to polarise the fusion chamber and increase cell fusion 5-fold. Optimisation of a direct current charge in combination with a fusogenic pulse in which fusion of a range of human cells approached or exceeded 30% of the total pulsed cells. The yield of hybrid prostate and breast cancer cells with dendritic cells was similar to the homologous cell fusion efficiencies indicating that dendritic cells were highly amenable to fusion with human tumour cells under similar electrical parameters. Elimination of unfused cells by density gradient and culture is possible to further increase the quantity of hybrid cells. The generation and purification of quantities of hybrid cells sufficient for human vaccination raises the possibility of rapid, autologous tumour antigen presenting vaccines for trial with common human tumours.
机译：由抗原呈递和肿瘤细胞融合产生的杂交细胞已显示出在啮齿动物癌症模型中诱导有效的保护性和治愈性抗肿瘤免疫力。杂交细胞疫苗在人类肿瘤治疗中的应用以及对疾病控制的及时干预受到要求获得足够的自体细胞来保存同源肿瘤抗原呈递的要求。在这项研究中，已经用多种人类造血，乳腺和前列腺细胞系研究了各种电融合方法在产生杂交人类细胞中的效率。使用电脉冲的细胞融合可通过多种刺激来增强，以电对齐细胞或使细胞接触。通过FACS分析，在来自Gene Pulser电穿孔设备的指数脉冲后，对细胞进行离心可提供最高产量的混合细胞杂种。电脉冲后在人细胞中产生的广泛的融合条件与这样的假设相矛盾，即先前的细胞接触对于细胞融合是必需的。并发的直流电中的细胞排列和聚乙二醇中细胞的渗透性扩张也产生了高水平的细胞融合。电穿孔比色皿的一个电极的打蜡作用使融合室极化，并使细胞融合增加了5倍。优化直流电和融合脉冲，在融合脉冲中，一系列人类细胞的融合达到或超过总脉冲细胞的30​​％。具有树突状细胞的前列腺和乳腺癌混合细胞的产量与同源细胞融合效率相似，这表明树突状细胞非常适合在相似的电参数下与人肿瘤细胞融合。通过密度梯度和培养消除未融合的细胞有可能进一步增加杂交细胞的数量。足以进行人类疫苗接种的杂交细胞数量的产生和纯化，增加了快速，自体的肿瘤抗原呈递疫苗用于普通人类肿瘤试验的可能性。

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