慢性粒细胞白血病患者PBMCs中CD4^＋CD25^＋细胞体外增殖及对CD4^＋CD25^-细胞增殖的影响  

In vitro proliferation of CD4^+CD25^+ T cells from PBMCs of the chronic myelocytic leukemia patients and their inhibitory effect on CD4^+CD25^- T cells

作　　者：陈伟[1] 曹东林[2] 王玲[2] 

机构地区：[1]广东省第二人民医院血液科,广东广州510317 [2]南方医科大学基因工程研究所,广东广州510515

出　　处：《南方医科大学学报》2008年第9期1610-1613,共4页Journal of Southern Medical University

基　　金：广州市科技计划(2005Z3-E0371)

摘　　要：目的探讨慢性粒细胞白血病(CML)患者外周血单个核细胞(PBMCs)来源的CD4+CD25+细胞体外增殖及对CD4+CD25-细胞增殖的影响。方法以免疫磁性分离方法(MACS)分选出CML患者外周血单个核细胞中的CD4+CD25+及CD4+CD25-细胞后,用流式细胞仪分析细胞的纯度及活力;再以小鼠抗人CD3单抗、小鼠抗人CD28单抗及rh IL-2作为刺激因子,CD4+CD25+细胞与CD4+CD25-细胞共培养,观察CD4+CD25+细胞对CD4+CD25-细胞增殖的抑制效应。结果(1)分选后健康对照组及CML患者PBMC中CD4+CD25+细胞纯度分别为(84.93±2.55)%、(86.32±2.40)%,两者相比,无显著差异(P>0.05);(2)经MACS分选后正常对照组与CML患者CD4+CD25+细胞活力分别为(98.12±0.68)%、(97.33±0.78)%,两者相比,无显著差异(P>0.05);(3)无论是健康对照还是CML患者的CD4+CD25+细胞均具有明显抑制效应性细胞如CD4+CD25-细胞的增殖,随着CD4+CD25+细胞数的增加,这种抑制增殖的能力也相应增加,当CD4+CD25+T∶CD4+CD25-T为1∶1时,抑制率最大。结论MACS分选法能够分选出高纯度及高活力的CD4+CD25+细胞,分选后CD4+CD25+细胞在体外均能抑制CD4+CD25-细胞增殖,且这种抑制效应呈一定效靶比关系。Objective To investigate the in vitro proliferation of CD4^＋CD25^＋ T cells from the peripheral blood mononuclear cells （PBMCs） of the chronic myelocytic leukemia patients and the inhibitory effect on CD4^＋CD25^- T cells. Methods Magnetic-activated cell sorting （MACS） was used to separate CD4^＋CD25^＋ T and CD4^＋CD25^- T cells from the PBMCs of patients with chronic myelocytic leukemia, and the purity and activity of CD4^＋CD25^＋ T cells were analyzed with flow cytometry. After stimulation with anti-CD3 mAb, anti-CD28 mAb and recombinant human interleukin-2 （rhIL-2）, the CD4^＋CD25^- and CD4^＋CD25^＋ T cells were cocultured to observe the inhibitory effect of CD4^＋CD25^＋ T on CD4^＋CD25^-T cells using MTT assay. Results After cell sorting, the purity of CD4^＋CD25^＋ T cells from healthy control and chronic myelocytic leukemia patients were （84.93±2.55）% and （86.32±2.40）%, respectively, showing no significant difference between them （P〉0.05）. The activity of CD4^＋CD25^＋and CD4^＋CD25^- T cells from healthy control and the leukemic patients was also comparable [（98.12±0.68）% vs （97.33±0.78）%, P〉0.05）. In the coculture, CD4^＋CD25^＋ T cells obviously inhibited CD4^＋CD25^- T cell proliferation in vitro, and the maximum inhibition occurred when CD4^＋CD25^＋T cells were cocultured with CD4^＋CD25^＋T cells at the ratio of 1：1. Conclusion The MACS system can effectively isolate CD4^＋CD25^＋ and CD4^＋CD25^- T cells.CD4^＋CD25^＋ T cells obviously inhibit the proliferation of CD4^＋CD25^-T cells in vitro, and the effect displays an effector-target ratio relationship.

关 键 词：慢性粒细胞白血病 CD4^+CD25^+细胞 细胞增殖 

分 类 号：R733.72[医药卫生—肿瘤]