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目的探讨在人重组的粒细胞集落刺激因子(rhG-CSF)动员的外周血单个核细胞中诱导产生CD4+CD25+调节性T细胞可行性及其表型和功能。方法收集本院外周血干细胞移植术供者rhG-CSF动员前外周血(PB组)和动员后的外周血采集物(G-PB组)各10例,用免疫磁珠法分选出CD4+CD25?T细胞,并用转化生长因子β1(TGF-β1)进行诱导,分别应用流式细胞术、RT-PCR和细胞增殖、抑制试验测定诱导后细胞的CD25、Foxp3表达和免疫抑制功能,比较2组之间CD4+CD25+T细胞转化率、抑制功能的差异。结果1)G-PB和PB来源的CD4+CD25?T细胞在抗-CD3 M cAb和TGF-β1作用下CD25+分子表达不同,分别为:(77.9±2.3)%和(65.7±4.2)%,差异有统计学意义(P<0.05);2)TGF-β1诱导产生的CD4+CD25+T细胞高表达Foxp3;3)PB、G-PB来源的TGF-β1诱导产生的CD4+CD25+T细胞具有免疫抑制功能,cpm值分别为:11 739±352和18 732±437(P<0.05)。结论G-CSF动员的外周血可作为CD4+CD25+调节性T细胞的重要来源。
Objective To investigate the feasibility, phenotype and function of CD4 + CD25 + regulatory T cells induced by human recombinant granulocyte colony stimulating factor (rhG-CSF) in peripheral blood mononuclear cells. Methods Peripheral blood stem cells (PB group) and mobilized peripheral blood samples (G-PB group) were collected from 10 recipients of peripheral blood stem cell transplantation in our hospital for 10 cases. CD4 + CD25 (superscript +) T cells and induced by transforming growth factor β1 (TGF-β1). Flow cytometry, RT-PCR and cell proliferation were used to detect the expression of CD25 and Foxp3, Differences in CD4 + CD25 + T cell transformation rates between groups and inhibition of function. Results The CD25 + expression of G-PB and PB-derived CD4 + CD25 + T cells under the action of anti-CD3 M cAb and TGF-β1 were 77.9 ± 2.3% and 65.7 ± 4.2%, respectively (P <0.05); 2) Foxp3 was highly expressed in CD4 + CD25 + T cells induced by TGF-β1; 3) PB + CD25 + T cells induced by TGF- Immunosuppressive function, cpm values were: 11 739 ± 352 and 18 732 ± 437 (P <0.05). Conclusion G-CSF-mobilized peripheral blood can serve as an important source of CD4 + CD25 + regulatory T cells.