Tan Huo, Zeng Lin-Juan, Ye Xu
Centre of Oncology and Hematology, The First Affiliated Hospital of Guangzhou Medical College, Guangzhou 510230, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2005 Jun;13(3):472-8.
To explore the possibility of in vitro induction of cord blood cell-derived lymphocytes into cytotoxic T lymphocytes (CTL) with anti-leukemia specificity, umbilical cord blood (UCB)-derived mononuclear cells were cultured with multiple cytokines to generate dendritic cells (DC) in vitro. Leukemia cells were irradiated with (137)Cs and activated by premature cytokines. The characteristics of maturation of DC were evaluated through morphology examination and flow cytometry. DC pulsed with leukemic antigens were co-cultured with lymphocytes. Cytotoxicity of the CTL to corresponding leukemic cells was measured with lactate dehydrogenase-release assay. The results showed that UCB-derived monocytes could be induced into typical DC in all of the 12 samples. Expression of immunological markers such as CD1a(+), HLA-DR(+), CD86(+), CD83(+) on DC were significantly up-regulated (P < 0.05). DC presenting leukemic antigens generated leukemia-specific CTL with a killing rate of (44.76 +/- 17.42)% at the E:T ratio of 50:1 against AML cells and a killing rate of (8.50 +/- 4.25)% at the E:T ratio of 50:1 against ALL cells. Whereas, these CTL present almost no killing effect on the mononuclear cells collected from the same patients in complete remission phase. It is concluded that (1) it is possible to induce UCB-derived monocytes into mature DC with typical morphology. (2) Cord blood derived mature DC presenting leukemia antigen can generate leukemia-specific CTL with vigorous cytotoxic activity against the same leukemia blasts and low killing activity against bone marrow cells of the same patients in complete remission phase.
为探讨体外诱导脐血细胞来源的淋巴细胞成为具有抗白血病特异性的细胞毒性T淋巴细胞（CTL）的可能性，将脐血（UCB）来源的单个核细胞与多种细胞因子共同培养，以在体外生成树突状细胞（DC）。白血病细胞用¹³⁷Cs照射并用早期细胞因子激活。通过形态学检查和流式细胞术评估DC的成熟特征。用白血病抗原脉冲处理的DC与淋巴细胞共培养。用乳酸脱氢酶释放试验测量CTL对相应白血病细胞的细胞毒性。结果显示，12个样本中的所有UCB来源的单核细胞均可被诱导成为典型的DC。DC上CD1a(+)、HLA-DR(+)、CD86(+)、CD83(+)等免疫标志物的表达显著上调（P < 0.05）。呈递白血病抗原的DC产生了白血病特异性CTL，在效靶比为50:1时对AML细胞的杀伤率为（44.76 ± 17.42）%，对ALL细胞在效靶比为50:1时的杀伤率为（8.50 ± 4.25）%。然而，这些CTL对处于完全缓解期的同一患者采集的单个核细胞几乎没有杀伤作用。结论为：（1）有可能将UCB来源的单核细胞诱导成为具有典型形态的成熟DC。（2）脐血来源的呈递白血病抗原的成熟DC可产生白血病特异性CTL，其对相同白血病原始细胞具有强烈的细胞毒性活性，而对处于完全缓解期的同一患者的骨髓细胞杀伤活性较低。
