Zhang Xiao-Hui, Xia Ling-Hui, Liu Zhong-Ping, Wei Wen-Ning, Hu Yu, Song Shan-Jun
Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2003 Feb;11(1):66-9.
The objective of this study is to investigate the effect of vaccination with dendritic cells pulsed with survivin antigen on activation of antileukemic T cells, and inhibiting proliferation of leukemic cells. The expression of survivin on acute leukemic cells were detected by cofocal microscopy and immunoprecipitation-Western blot. DCs collected from peripheral blood mononuclear cells were pulsed with survivin purified proteins. Stimulation index (SI) and antileukemia CTL induction were analyzed with (3)H-TdR incorporation and (51)Cr releasing assay, respectively. The phenotype of T cells and DCs were identified by flow cytometry. By immunofluorescence of bone marrow and peripheral blood mononuclear cells, survivin expression was detected in 16 out of 19 AML cases (84.2%). The results showed that survivin fluorescence distribution was in cytoplasm. DCs from peripheral blood mononuclear cells were successfully induced, with typical DC morphologic characteristic. The vaccination with dendritic cells pulsed with survivin antigen dramatically stimulated the proliferation of T cells. The DCs loading survivin activated T cells with higher CD4(+) T(H) ratio as compared with DCs group, T cells activated with DCs expressed CD8 and CD56. Survivin DCs significantly inhibited the growth of leukemic cells in vitro. In conclusion, survivin antigen expressed in the cytoplasm of leukemic cells, leukemic vaccination with DCs pulsed with survivin antigen in vitro inhibited the proliferation of leukemic cells, that may be a pathway for therapy of leukemia.
本研究的目的是探讨用survivin抗原脉冲处理的树突状细胞进行疫苗接种对抗白血病T细胞激活及抑制白血病细胞增殖的影响。通过共聚焦显微镜和免疫沉淀-蛋白质印迹法检测急性白血病细胞上survivin的表达。用survivin纯化蛋白脉冲处理从外周血单个核细胞收集的树突状细胞。分别用³H-TdR掺入法和⁵¹Cr释放试验分析刺激指数（SI）和抗白血病CTL诱导情况。通过流式细胞术鉴定T细胞和树突状细胞的表型。通过骨髓和外周血单个核细胞的免疫荧光检测，19例急性髓系白血病（AML）病例中有16例（84.2%）检测到survivin表达。结果显示survivin荧光分布于细胞质中。成功诱导出具有典型树突状细胞形态特征的外周血单个核细胞来源的树突状细胞。用survivin抗原脉冲处理的树突状细胞进行疫苗接种显著刺激了T细胞的增殖。与树突状细胞组相比，负载survivin的树突状细胞激活的T细胞具有更高的CD4⁺ Th比例，用树突状细胞激活的T细胞表达CD8和CD56。survivin树突状细胞在体外显著抑制白血病细胞的生长。总之，白血病细胞细胞质中表达的survivin抗原，体外用survivin抗原脉冲处理的树突状细胞进行白血病疫苗接种可抑制白血病细胞的增殖，这可能是白血病治疗的一条途径。
