Department for Hematopoetic Cell Transplantation, Med. Dept.3, Workgroup Immunomodulation, University Hospital of Munich, Munich.
Department for Hematology and Oncology, Municipal Hospital, Oldenburg.
J Immunother. 2019 Jun;42(5):143-161. doi: 10.1097/CJI.0000000000000266.
Strategies to stabilize remissions by specific elimination of residual acute myeloid leukemia (AML) blasts are needed. Leukemia-derived dendritic cell (DCleu/DC) generated from myeloid blasts improve antileukemic T-cell reactivity and install T-cell memory. Interferon (IFN)α-DC methods produce DCleu from chronic myeloid leukemia-patients (pts') blood. Various INFα-containing versus other DC methods were studied to produce DCleu (evaluated by flowcytometry) from AML-pts' blast-containing mononuclear (MNC) or whole blood (WB). After DCleu/DC stimulation in mixed lymphocyte cultures, T cells' potential to gain antileukemic cytotoxicity was studied and correlated with different DC methods and DCleu/DC counts. (1) Generation of DCleu/DC: (a) "IFN-GIT" [containing granulocyte macrophage-colony stimulating factor (GM-CSF)+IFNα+ tumor necrosis factor (TNF)-α] produced DC successfully (≥10% DC, ≥5% DCleu/cells) from AML-MNC (WB) in 54 (56%), "MCM-Mimic" in 76 (75%), "Picibanil" in 83 (64%), and "Calcium-ionophore" in 42 (67%) of cases. Proportions of DC subtypes in MNC (WB) were comparable with all DC methods, (b) IFNα combinations containing only GM-CSF+IFNα or only IFNα showed low efficiency to produce DCleu/DC from MNC (WB) compared with "IFN-GIT." (2) Antileukemic functionality: DCleu/DC-stimulated T cells showed improved leukemia cytotoxicity compared with blast cells or unstimulated T cells. The highest blast proliferation (=insufficient T cells) was seen with "IFN-GIT" DC-stimulated T cells. Probability to respond to immunotherapy or to obtain blast lysis of DC-stimulated T cells correlated with high proportions of DCleu/DC after DC culture, independent of DC-generating methods. (3) Cytokine release profiles: levels of interleukin-6, IFN-γ, and interleukin-2 were significantly lower in DC culture supernatants (from MNC/WB) with "IFN-GIT" compared with "MCM," "Pici," and "Ca" DC supernatants. Our data show that (1) WB culture simulates AML-pts' in vivo situation, (2) DC generation is possible from AML-MNC (WB) with IFNα-containing and other DC methods, (3) successful IFNα-DC generation needs GM-CSF+IFNα+TNF-α (IFN-GIT); however, "IFN-GIT" produces less DCleu/DC compared with other (non-IFNα) DC methods, (4) T cells stimulated with "IFN-GIT"-produced DCleu/DC yielded comparable antileukemic cytotoxicity; however, in cases without achieved blast lysis, an increased blast proliferation was observed.
需要通过特异性清除残留急性髓系白血病 (AML) blasts 来稳定缓解。从髓系 blasts 中生成的白血病衍生树突状细胞 (DCleu/DC) 可提高抗白血病 T 细胞反应性并建立 T 细胞记忆。干扰素 (IFN)α-DC 方法可从慢性髓系白血病患者 (pts') 的血液中产生 DCleu。研究了各种含有 IFNα的 DC 方法与其他 DC 方法，以从 AML-pts 的含 blast 的单核细胞 (MNC) 或全血 (WB) 中产生 DCleu (通过流式细胞术评估)。在混合淋巴细胞培养中对 DCleu/DC 进行刺激后，研究了 T 细胞获得抗白血病细胞毒性的潜力，并与不同的 DC 方法和 DCleu/DC 计数相关联。
(1)DCleu/DC 的生成：(a)“IFN-GIT”[含有粒细胞巨噬细胞集落刺激因子 (GM-CSF)+IFNα+肿瘤坏死因子 (TNF)-α]成功地从 AML-MNC (WB) 中生成了 DC (≥10% DC，≥5% DCleu/细胞) (56%)，“MCM-Mimic”为 76% (75%)，“Picibanil”为 83% (64%)，“钙离子载体”为 42% (67%)。在 MNC (WB) 中，所有 DC 方法的 DC 亚型比例均相似，(b)仅包含 GM-CSF+IFNα或仅包含 IFNα 的 IFNα 组合与“IFN-GIT”相比，从 MNC (WB) 中生成 DCleu/DC 的效率较低。
(2)抗白血病功能：与 blast 细胞或未刺激的 T 细胞相比，DCleu/DC 刺激的 T 细胞显示出改善的白血病细胞毒性。在用“IFN-GIT”刺激的 T 细胞中观察到最高的 blast 增殖(=T 细胞不足)。与“IFN-GIT”DC 刺激的 T 细胞相比，对免疫疗法有反应或获得 DC 刺激的 T 细胞对 blast 的溶解的可能性与 DC 培养后 DCleu/DC 的高比例相关，而与生成 DC 的方法无关。
(3)细胞因子释放谱：与“MCM”、“Pici”和“Ca”DC 上清液相比，“IFN-GIT”上清液中的白细胞介素-6、IFN-γ 和白细胞介素-2 水平明显较低。我们的数据表明：(1)WB 培养模拟了 AML-pts 的体内情况，(2)可以从 AML-MNC (WB) 中用含有 IFNα 和其他 DC 方法生成 DC，(3)成功的 IFNα-DC 生成需要 GM-CSF+IFNα+TNF-α(IFN-GIT)；然而，与其他 (非 IFNα) DC 方法相比，“IFN-GIT”产生的 DCleu/DC 较少，(4)用“IFN-GIT”产生的 DCleu/DC 刺激的 T 细胞产生了相当的抗白血病细胞毒性；然而，在未达到 blast 溶解的情况下，观察到 blast 增殖增加。
