抗CD25单抗对肾移植患者调节性T细胞生存和功能改变影响的研究

发布时间：2018-10-13 19:55

【摘要】：第一部分抗CD25单抗诱导治疗对肾移植早期患者外周血调节性T细胞及其亚群的影响目的本部分拟研究肾移植早期患者接受抗CD-25单克隆抗体诱导治疗后对其外周血调节性T细胞(Treg)及其亚群的影响。方法自2012至2013年间,10名肾移植患者根据是否接受巴利昔单抗诱导治疗,被分为诱导组和非诱导组,分别在肾移植术前两小时、术后1周、术后2周采集外周血液样本,使用流式细胞仪分析其调节性T细胞及其亚群的比例。结果与非诱导组相比,巴利昔单抗诱导组外周血中CD25+T细胞和CD25+Foxp3+双标记T细胞均出现明显的下降,并在术后两周内保持较低水平。活化型Treg与分泌细胞因子型Treg亚群也在术后1周内出现了短暂的下降。抑制性转录因子FOXP3并没有在接受诱导治疗后受到显著影响。结论在肾移植早期阶段,抗CD25单克隆抗体下调外周血调节性T细胞及其活化亚群的比例,但其免疫抑制功能未受到明显影响。第二部分体外分离培养调节性T细胞后,抗CD25单抗对其生存和功能的影响目的本部分拟在体外分离培养健康志愿者外周血PBMC调节性T细胞,研究抗CD25单抗对其生存和免疫抑制功能的影响。方法分离健康志愿者外周血PBMC,采用流式分选方法分选CD4+CD25+CD 127-调节性T细胞,加入刺激因子与之共培养扩增,使用抗CD25单克隆抗体处理后,使用AnnexinV/PI双染法检测其凋亡情况,使用ELISA方法检测其IL-10, TGF-β等细胞因子的分泌情况。结果与对照组相比,抗CD25单抗处理组在细胞凋亡、细胞因子分泌等方面未出现明显差异。结论在体外实验中,使用抗CD25单克隆抗体后,调节性T细胞的生存及其免疫抑制因子的分泌功能并未受到明显影响。
[Abstract]:Effect of anti CD25 monoclonal antibody induction therapy on peripheral blood regulatory T cells and its subsets in patients with renal transplantation objective to study the effect of anti CD-25 monoclonal antibody induction therapy on peripheral blood of early renal transplantation patients Effects of regulatory T cell (Treg) and its subsets. Methods from 2012 to 2013, 10 renal transplant patients were divided into two groups according to whether they received bariximab induction therapy or not. Peripheral blood samples were collected two hours before renal transplantation, one week after transplantation and two weeks after renal transplantation. The proportion of regulatory T cells and their subsets was analyzed by flow cytometry. Results compared with the control group, the levels of CD25 T cells and CD25 Foxp3 double labeled T cells in the peripheral blood of the Balicicab induced group were significantly lower than those in the non-induction group, and remained at a low level within two weeks after operation. The subsets of activated Treg and secreting cytokine type Treg also decreased 1 week after operation. Inhibitory transcription factor FOXP3 was not significantly affected after induction therapy. Conclusion in the early stage of renal transplantation, monoclonal antibodies against CD25 down-regulate the proportion of peripheral blood regulatory T cells and their activated subsets, but their immunosuppressive function is not significantly affected. The second part: effects of anti CD25 monoclonal antibody on survival and function of regulatory T cells isolated and cultured in vitro objective to isolate and culture PBMC regulatory T cells from peripheral blood of healthy volunteers in vitro. To study the effect of anti CD25 monoclonal antibody on its survival and immunosuppressive function. Methods Peripheral blood PBMC, was isolated from healthy volunteers and CD4 CD25 CD 127-regulatory T cells were separated by flow sorting method, and co-cultured and amplified by adding stimulating factor. After treated with anti-CD25 monoclonal antibody, the apoptosis of CD4 CD25 CD 127-regulatory T cells was detected by AnnexinV/PI double staining method. ELISA method was used to detect the secretion of IL-10, TGF- 尾 and other cytokines. Results there was no significant difference in apoptosis and cytokine secretion between anti CD25 McAb group and control group. Conclusion in vitro, the survival of regulatory T cells and the secretion of immunosuppressive factors were not significantly affected by the use of monoclonal antibodies against CD25.
【学位授予单位】：复旦大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R699.2

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