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目的 探讨自体白血病细胞裂解物 (ACL)冲击的完全缓解期的急性髓系白血病 (AML CR)患者骨髓细胞衍生的树突状细胞 (DC)体外能否刺激自体T细胞产生特异性抗AML细胞的细胞毒活性。方法 应用羊红细胞玫瑰花结程序从AML CR患者的骨髓中分离出去T细胞的骨髓单个核细胞 (TD BMNC) ,并培养在含联合细胞因子 (GM CSF、IL 4、SCF或TNF α)的条件下以产生DC ,并在培养的第 5天用ACL进行冲击。培养 7d后收获细胞 ,用流式细胞仪测定其成熟DC表型。同时 ,这些细胞与经抗CD3抗体激活过的自体T细胞在低浓度IL 2条件下共培养 7d ,以产生细胞毒性T淋巴细胞(CTL)。用乳酸脱氢酶释放实验测定溶细胞活性。结果 1 2例AML CR患者培养的骨髓单个核细胞均分化为成熟DC。其中 6例完成CTL活性实验。当效∶靶 =2 0∶1时 ,ACL冲击的DC致敏的自体T细胞与单纯IL 2或IL 2加无抗原冲击的DC组比较对自体AML细胞有明显的杀伤活性 ,而对K562细胞均无明显影响 (P <0 .0 1 )。结论 用AML CR患者白细胞裂解物冲击的骨髓细胞衍生的DC体外致敏自体T细胞可以产生AML细胞相关抗原特异性的CTL。
Objective To investigate whether bone marrow cell-derived dendritic cells (DCs) from patients with acute myeloid leukemia (AML CR) undergoing complete remission of autologous leukemic cell lysate (ACL) shock can stimulate autologous T cells to produce specific anti-AML cells in vitro. Cytotoxic activity. METHODS: The bone marrow mononuclear cells (TD BMNC) of T cells were isolated from the bone marrow of AML CR patients using the sheep red blood cell rosette procedure, and cultured in conditions containing the combined cytokines (GM CSF, IL 4, SCF, or TNF α). Next to produce DC, and on the fifth day of culture with ACL shock. Cells were harvested 7 days after culture and their mature DC phenotypes were determined by flow cytometry. At the same time, these cells were co-cultured with anti-CD3 antibody-activated autologous T cells at low concentrations of IL 2 for 7 days to produce cytotoxic T lymphocytes (CTLs). The lactate dehydrogenase release assay was used to determine cytolytic activity. Results One hundred and twenty bone marrow mononuclear cells cultured in patients with AML CR all differentiated into mature DCs. Six of them completed the CTL activity experiment. When the effect: target = 2 0:1, the DC-sensitized autologous T cells impacted by ACL showed significant killing activity against autologous AML cells compared with DCs with IL-2 or IL-2 alone and antigen-free DCs, but against K562 cells. No significant effect (P <0. 0 1). Conclusions In vitro sensitization of autologous T cells with DCs derived from leukocyte lysate in patients with AML CR can generate AML cell-specific antigen-specific CTLs.