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Clinical reagents of GM-CSF and IFN-α induce the generation of functional chronic myeloid leukemia dendritic cells in vitro

机译:GM-CSF和IFN-α的临床试剂在体外诱导功能性慢性髓性白血病树突状细胞的生成

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Dendritic cells (DCs) have been successfully induced in vitro from chronic myeloid leukemia (CML) cells, which may provide a promising immunotherapeutic protocol for CML. To facilitate the optimization of DCs-based vaccination protocols, we investigated the efficiency of in vitro generation of DCs from bone marrow mononuclear cells of CML patients by clinical reagents of GM-CSF and IFN-α. Bone marrow mononuclear cells were isolated from eight CML patients and CML-DCs were generated in the presence of different cytokines (Group A: GM-CSF for research and IL-4 for research; Group B: GM-CSF for injection and IFN-α for injection) in RMPI-1640 medium containing 10% human AB serum. After 8?days, the morphologic features of CML-DCs were observed and their immunophenotypes were analyzed by flow cytometry. The activity of CML-DCs was determined by evaluating their ability to stimulate allogeneic mixed lymphocyte reaction (allo-MLR) and anti-leukemic cytotoxic T lymphocytes (CTLs). The culture protocols were successful in generating functional CML-DCs from all the CML patients as evidenced by the significant upregulation of CD80, CD86, CD83 HLA-DR and CD1a compared to pre-cultured (p?
机译:树突状细胞(DCs)已成功地从慢性粒细胞白血病(CML)细胞体外诱导,这可能为CML提供有希望的免疫治疗方案。为了促进基于DC的疫苗接种方案的优化,我们研究了通过GM-CSF和IFN-α的临床试剂从CML患者的骨髓单个核细胞体外生成DC的效率。从八名CML患者中分离出骨髓单个核细胞,并在存在不同细胞因子的情况下产生CML-DC(A组:用于研究的GM-CSF和IL-4; B组:用于注射的GM-CSF和IFN-α注射用)在含有10%人源AB血清的RMPI-1640培养基中。 8天后,观察CML-DC的形态特征,并通过流式细胞仪分析其免疫表型。通过评估它们刺激同种异体混合淋巴细胞反应(allo-MLR)和抗白血病细胞毒性T淋巴细胞(CTL)的能力来确定CML-DC的活性。培养方案成功地从所有CML患者中生成了功能性CML-DC,这与CD80,CD86,CD83 HLA-DR和CD1a与预培养相比显着上调(p <0.05)和同种异体T升高证明了这一点。细胞刺激增殖能力(p≤0.05)。 CML-DC可以刺激特定的抗白血病反应。总而言之,我们证明了临床试剂GM-CSF和IFN-α的结合诱导了DC的产生,这些DC具有在体外刺激特定抗白血病CTL反应的能力,表明它们可用于CML患者的临床疫苗接种方案。

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