Apoptosis of monocytes and the influence on yield of monocyte-derived dendritic cells
Apoptosis of monocytes and the influence on yield of monocyte-derived dendritic cells
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DOI：
10.1016/j.jim.2004.08.010
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发表时间：
2004-11-01
影响因子：
2.2
通讯作者：
Noll, T
中科院分区：
文献类型：
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作者：
Bohnenkamp, HR;Burchell, JM;Noll, T
单核细胞来源的树突状细胞（DC）目前正在广泛评估作为细胞疫苗用于癌症治疗。大规模电池产品的需求需要优化和标准化的协议。然而，据报道，来自接种的单核细胞的DC的产率总是低于50%。在本研究中，我们研究了这种细胞损失是否是由接种的单核细胞的起始群体的性质引起的。通过免疫磁珠选择富集CD 14细胞，并通过膜联蛋白V/碘化丙啶测定分析细胞凋亡。我们发现，37.8 +/- 11.1%（n=8）的新鲜分离的单核细胞从血沉棕黄层的健康供体经历程序性细胞死亡。进一步分析凋亡细胞在分化过程中的命运表明吞噬作用。用GM-CSF和白细胞介素-4将单核细胞分化成存活的、非凋亡的未成熟树突状细胞群体。添加肿瘤坏死因子-α和前列腺素E-2导致完全成熟的树突状细胞，其通过表型分析和同种异体和MHC I类限制性T细胞应答进行评价。约90.2 +/-16.7%的非凋亡单核细胞群分化为活的成熟树突状细胞。这些结果表明，树突状细胞的产量主要受接种物中凋亡细胞百分比的影响，这对临床应用中DC的产生具有影响。(C)2004 Elsevier B. V.保留所有权利。
Monocyte-derived dendritic cells (DC) are currently under extensive evaluation as cell vaccines for cancer treatment. The requirement for large-scale cell products demands optimized and standardized protocols. However, the yield of DCs from inoculated monocytes is reported to be always lower than 50%. In this present study we investigated whether this cell loss was caused by the properties of the starting population of inoculated monocytes. CD14 cells were enriched by immunomagnetic-bead selection and analyzed for apoptosis by an annexin V/propidium iodide assay. We found that 37.8 +/- 11.1% (n=8) of freshly isolated monocytes from buffy coats of healthy donors underwent programmed cell death. Further analysis of the fate of apoptotic cells during differentiation suggested phagocytosis. Monocytes were differentiated with GM-CSF and interleukin-4 into a viable, non-apoptotic population of immature dendritic cells. Addition of tumor necrosis factor-alpha and prostaglandin E-2 resulted in fully matured dendritic cells, which were evaluated by phenotypic analysis and by allogeneic and MHC class-I-restricted T-cell responses. About 90.2 +/- 16.7% of the non-apoptotic monocyte population differentiated to viable matured dendritic cells. These results indicate that the yield of dendritic cells is mainly influenced by the percentage of apoptotic cells in the inoculum, and this has implications for DC generation in clinical applications. (C) 2004 Elsevier B.V. All rights reserved.