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[脐带血来源的CD8 + 细胞毒性T淋巴细胞对白血病的特异性抗白血病效应诱导]

[Specific induction of anti-leukemia effects by umbilical cord cell-derived CD8+ T cytotoxic lymphocytes].

作者信息

Liu Xin, Tan Huo, Wang Chun-Yan, Huang Zhen-Qian, Zhang Huan-Zhu

机构信息

Centre of Oncology and Hematology, First Affiliated Hospital of Guangzhou Medical College, Guangzhou 510230, China.

出版信息

Zhonghua Xue Ye Xue Za Zhi. 2006 Jul;27(7):452-5.

Abstract

OBJECTIVE

To explore the specific anti-leukemia immune response of CD8+ cytotoxic T lymphocyte (CTL) derived from cord blood (CB) ex vivo and evaluate the feasibilities and values of the CTL for specific immunotherapy.

METHODS

Dendritic cells (DC) were induced from mononuclear cells (MNC) by combination cytokines in 10 CB samples. Loading U937 cell lysate antigen on the mature DC, they could stimulate the lymphocytes of the same origin to generate CTL. MidiMACS was used to isolate CD8+ CTL. Analysis of DC was performed by inverted microscopy, scanning electron microscopy and flow cytometry. Methyl thiazolyl tetrazolium (MTT) assay was used to evaluate the cytotoxicity of the CTL.

RESULTS

Cocultured with GM-CSF, IL-4, TNF-alpha and PGE2, CB-MNC could be induced into functional DC with typical morphology. The mean cytotoxicity of CD8+ CTL to U937 cells was significant stronger than that of CD8- CTL and TL at the same E: T ratios. The mean cytotoxicity rate of CD8+ CTL to U937 cells was higher than that to K562 cells [(66.36 +/- 12.43)% vs (41.97 +/- 14.24)%] at E: T ratio of 40: 1 (P < 0.05). The cytotoxicity of CD8- CTL to K562 cells showed no difference from that to U937 cells (P > 0.05).

CONCLUSION

Mature CB-DC loading U937 cell antigens could induce CB-T lymphocytes to generate leukemia-specific CD8+ CTL. The cytotoxicity of the CD8+ CTL is specific against U937 cells and is more potent than that of CD8- CTL.

摘要

目的

探讨体外从脐血(CB)中获得的CD8+细胞毒性T淋巴细胞(CTL)的特异性抗白血病免疫反应,并评估CTL用于特异性免疫治疗的可行性和价值。

方法

采用联合细胞因子从10份CB样本的单核细胞(MNC)中诱导树突状细胞(DC)。将U937细胞裂解物抗原负载于成熟DC上,可刺激同源淋巴细胞产生CTL。使用MidiMACS分离CD8+CTL。通过倒置显微镜、扫描电子显微镜和流式细胞术对DC进行分析。采用甲基噻唑基四氮唑(MTT)法评估CTL的细胞毒性。

结果

与GM-CSF、IL-4、TNF-α和PGE2共培养,CB-MNC可被诱导为具有典型形态的功能性DC。在相同的效靶比下,CD8+CTL对U937细胞的平均细胞毒性明显强于CD8-CTL和总淋巴细胞(TL)。在效靶比为40:1时,CD8+CTL对U937细胞的平均细胞毒性率高于对K562细胞的毒性率[(66.36±12.43)%对(41.97±14.24)%](P<0.05)。CD8-CTL对K562细胞的细胞毒性与对U937细胞的细胞毒性无差异(P>0.05)。

结论

负载U937细胞抗原的成熟CB-DC可诱导CB-T淋巴细胞产生白血病特异性CD8+CTL。CD8+CTL的细胞毒性对U937细胞具有特异性,且比CD8-CTL更强。

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