Medical Department III, University Hospital Grosshadern, Ludwig-Maximilians-University, 81377 Munich, Germany.
J Immunother. 2010 Feb-Mar;33(2):185-99. doi: 10.1097/CJI.0b013e3181b8f4ce.
Myeloid-leukemic cells (AML, MDS, CML) can be differentiated to leukemia-derived dendritic cell [DC (DCleu)] potentially presenting the whole leukemic antigen repertoire without knowledge of distinct leukemia antigens and are regarded as promising candidates for a vaccination strategy. We studied the capability of 6 serum-free DC culture methods, chosen according to different mechanisms, to induce DC differentiation in 137 cases of AML and 52 cases of MDS. DC-stimulating substances were cytokines ("standard-medium", "MCM-Mimic", "cytokine-method"), bacterial lysates ("Picibanil"), double-stranded RNA ["Poly (I:C)"] or a cytokine bypass method ("Ca-ionophore"). The quality/quantity of DC generated was estimated by flow cytometry studying (co) expressions of "DC"antigens, costimulatory, maturation, and blast-antigens. Comparing these methods on average 15% to 32% DC, depending on methods used, could be obtained from blast-containing mononuclear cells (MNC) in AML/MDS cases with a DC viability of more than 60%. In all, 39% to 64% of these DC were mature; 31% to 52% of leukemic blasts could be converted to DCleu and DCleu-proportions in the suspension were 2% to 70% (13%). Average results of all culture methods tested were comparable, however not every given case of AML could be differentiated to DC with 1 selected method. However performing a pre-analysis with 3 DC-generating methods (MCM-Mimic, Picibanil, Ca-ionophore) we could generate DC in any given case. Functional analyses provided proof, that DC primed T cells to antileukemia-directed cytotoxic cells, although an anti-leukemic reaction was not achieved in every case. In summary our data show that a successful, quantitative DC/DCleu generation is possible with the best of 3 previously tested methods in any given case. Reasons for different functional behaviors of DC-primed T cells must be evaluated to design a practicable DC-based vaccination strategy.
髓系白血病细胞（AML、MDS、CML）可以分化为白血病衍生的树突状细胞[DC（DCleu）]，能够呈现整个白血病抗原库，而无需了解独特的白血病抗原，被认为是疫苗接种策略的有前途的候选者。我们研究了根据不同机制选择的 6 种无血清 DC 培养方法在 137 例 AML 和 52 例 MDS 中的诱导 DC 分化的能力。DC 刺激物是细胞因子（“标准培养基”、“MCM-Mimic”、“细胞因子方法”）、细菌裂解物（“Picibanil”）、双链 RNA [“Poly（I：C）”] 或细胞因子旁路方法（“钙离子载体”）。通过流式细胞术研究“DC”抗原、共刺激、成熟和原始细胞抗原的（共）表达来评估生成的 DC 的质量/数量。将这些方法进行比较，平均可以从 AML/MDS 病例的含原始细胞的单核细胞（MNC）中获得 15%至 32%的 DC，具体取决于使用的方法，并且 DC 存活率超过 60%。总之，39%至 64%的这些 DC 是成熟的；31%至 52%的白血病原始细胞可以转化为 DCleu，并且悬浮液中的 DCleu 比例为 2%至 70%（13%）。所有测试的培养方法的平均结果相当，但是并非每个 AML 病例都可以用 1 种选定的方法分化为 DC。然而，通过对 3 种生成 DC 的方法（MCM-Mimic、Picibanil、钙离子载体）进行预分析，我们可以在任何给定的病例中生成 DC。功能分析证明，DC 可以使 T 细胞对白血病定向细胞毒性细胞产生免疫反应，尽管并非在每个病例中都能实现抗白血病反应。总之，我们的数据表明，通过在任何给定的病例中使用之前测试的 3 种最佳方法之一，可以成功地进行定量的 DC/DCleu 生成。必须评估 DC 激活的 T 细胞的不同功能行为的原因，以设计可行的基于 DC 的疫苗接种策略。
