Efficient activation of autologous tumor-specific T cells: A simple coculture technique of autologous dendritic cells compared to established cell fusion strategies in primary human colorectal carcinoma
Efficient activation of autologous tumor-specific T cells: A simple coculture technique of autologous dendritic cells compared to established cell fusion strategies in primary human colorectal carcinoma
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DOI：
10.1097/cji.0b013e31802bfefe
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发表时间：
2007-05-01
影响因子：
3.9
通讯作者：
Wolf, Juergen
中科院分区：
文献类型：
--
作者：
Draube, Andreas;Beyer, Marc;Wolf, Juergen
不同的技术已经被用于将整个肿瘤抗原（TAs）传递到树突状细胞（dc），然后呈递给T细胞。这些包括全肿瘤rna转染的DC，装载肿瘤来源的凋亡小体或肿瘤细胞裂解物的DC的制备，以及DC肿瘤细胞融合。使用这种技术进行了早期临床试验。本研究旨在重新审视肿瘤细胞操作在基于dc的结直肠癌免疫治疗策略中的必要性。我们研究了一种简单的自体单核细胞来源的dc和人原发性结直肠癌（pCC）共培养方法，比较了两种已知的细胞融合策略对自体T细胞摄取、加工和呈递TAs的效果。在共培养或融合之前，pCC已冷冻保存，无需进一步操作。采用荧光显微镜和荧光染料标记细胞流式细胞术监测DCs对pCC的吞噬。共培养过程导致双阳性细胞比例高达22%，因此与细胞融合后观察到的结果相当。更重要的是，DC与自体pCC共培养后，在与DC/pCC融合的相同数量的患者中诱导了显著的肿瘤特异性干扰素- γ产生的自体T细胞。此外，通过与pCC共培养的dc刺激产生肿瘤特异性主要组织相容性复合体I类限制性细胞毒性T淋巴细胞。在先前的人类癌症研究中，共培养技术被认为是较差的。相反，我们的数据强烈表明，至少对于人类pCC和自体dc，这种简单的共培养方法与现有的融合技术相比同样有效。
Different technologies have been employed to deliver the whole spectrum of tumor antigens (TAs) to dendritic cells (DCs) to be presented to T cells. These include whole tumor RNA-transfected DCs, preparations of DCs loaded with tumor-derived apoptotic bodies or tumor cell lysates, and DC tumor cell fusions. Early clinical trials have been conducted using such techniques. The presented study was aimed to revisit the necessity of tumor cell manipulation in DC-based immunotherapy strategies for colorectal carcinoma. We investigated a simple coculture method of autologous monocyte-derived DCs and human primary colorectal carcinoma (pCC) in comparison with 2 well-described cell fusion strategies for the efficacy of uptake, processing and presentation of TAs to autologous T cells. Before coculture or fusion, pCC had been cryopreserved without further manipulation. Fluorescence microscopy and flow cytometry analyses of fluorescent dye labeled cells were used for monitoring engulfment of pCC by DCs. The coculture procedure resulted in a double positive cell fraction of up to 22%, and thus was comparable to that observed after cell fusion. More important, DCs after coculture with autologous pCC induced significant tumor-specific interferon-gamma-producing autologous T cells in the same number of patients as DC/pCC fusions. Furthermore, tumor-specific major histocompatibility complex class I restricted cytotoxic T lymphocytes were generated by stimulation with DCs cocultured with pCC. In prior Studies for human carcinomas coculture techniques were described to be inferior. In contrast, our data strongly suggest that at least for human pCC and autologous DCs this simple coculture method is similarly efficient compared to established fusion techniques.