Controlled-rate freezer cryopreservation of highly concentrated peripheral blood mononuclear cells results in higher cell yields and superior autologous T-cell stimulation for dendritic cell-based immunotherapy.
Controlled-rate freezer cryopreservation of highly concentrated peripheral blood mononuclear cells results in higher cell yields and superior autologous T-cell stimulation for dendritic cell-based immunotherapy.
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DOI：
10.1007/s00262-012-1262-0
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发表时间：
2012-11
影响因子：
5.8
通讯作者：
Haenssle, Holger A.
中科院分区：
文献类型：
--
作者：
Buhl, Timo;Legler, Tobias J.;Rosenberger, Albert;Schardt, Anke;Schoen, Michael P.;Haenssle, Holger A.
大量功能有效的树突状细胞(DC)的可获得性是感染性或恶性疾病免疫治疗试验的主要挑战之一。在临床试验中，DC数量少或T细胞活化不足可能导致过早终止治疗和不满意的免疫反应。基于单核细胞的冷冻保存在所产生的DC数量和免疫刺激能力方面优于未成熟或成熟DC的观点，我们旨在建立一种用于DC免疫治疗的高度浓缩外周血单个核细胞(PBMC)的冷冻保存的优化方案。对冷冻保存的细胞制剂进行定量回收、活性、表型和功能特性分析。与标准的异丙醇(IPA)冷冻相比，将PBMC冷冻保存在自动控制速率冰柜(CRF)中并随后解冻和分化可显著提高未成熟和成熟DC的细胞产量。使用CRF后的未成熟DC产量和总蛋白含量与新鲜制备的PBMC的结果相当，比标准IPA冷冻的结果高出约50%。虽然分化标志物、同种异体T细胞刺激、活性和细胞因子谱与标准冷冻程序中的DC相似，但从CRF冷冻保存的PBMC产生的DC诱导自体效应T细胞释放显著更高的抗原特异性干扰素-γ。综上所述，自动控制率冷冻高浓度PBMC代表了一种提高DC产量和自体T细胞刺激的改进方法。
Availability of large quantities of functionally effective dendritic cells (DC) represents one of the major challenges for immunotherapeutic trials against infectious or malignant diseases. Low numbers or insufficient T-cell activation of DC may result in premature termination of treatment and unsatisfying immune responses in clinical trials. Based on the notion that cryopreservation of monocytes is superior to cryopreservation of immature or mature DC in terms of resulting DC quantity and immuno-stimulatory capacity, we aimed to establish an optimized protocol for the cryopreservation of highly concentrated peripheral blood mononuclear cells (PBMC) for DC-based immunotherapy. Cryopreserved cell preparations were analyzed regarding quantitative recovery, viability, phenotype, and functional properties. In contrast to standard isopropyl alcohol (IPA) freezing, PBMC cryopreservation in an automated controlled-rate freezer (CRF) with subsequent thawing and differentiation resulted in significantly higher cell yields of immature and mature DC. Immature DC yields and total protein content after using CRF were comparable with results obtained with freshly prepared PBMC and exceeded results of standard IPA freezing by approximately 50 %. While differentiation markers, allogeneic T-cell stimulation, viability, and cytokine profiles were similar to DC from standard freezing procedures, DC generated from CRF-cryopreserved PBMC induced a significantly higher antigen-specific IFN-γ release from autologous effector T cells. In summary, automated controlled-rate freezing of highly concentrated PBMC represents an improved method for increasing DC yields and autologous T-cell stimulation.
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影响因子：
6.4
作者：
Elkord, E;Williams, PE;Rowbottom, AW
通讯作者：
Rowbottom, AW
影响因子：
3.9
作者：
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56.9
作者：
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通讯作者：
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158.5
作者：
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通讯作者：
Young, J.
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4.2
作者：
Breckpot, K;Corthals, J;Thielemans, K
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Thielemans, K