脐血单个核细胞CD25 mRNA表达的研究

Expression of CD25 mRNA in cord blood mononuclear cells

目的:观察外周血、脐血CD25 mRNA和CD25表达水平,初步了解CD25在幼稚T细胞表达中的作用。方法:以F icoll-Hypaque分离30名健康产妇外周血单个核细胞(PBMC)及脐血单个核细胞(CBMC);以生物素-链霉亲和素(BSA)法检测PHA诱导前后CD25表达水平;ELISA法测定血清sIL-2R含量;Real-tim e PCR检测CD25 mRNA含量。结果:静息状态和PHA诱导状态CD25(%)、血清中的sIL-2R(pg/ml)及CD25mRNA(logcDNA/logGAPDH)表达水平分别为在孕妇外周血中和在脐血中均有显著差异(P<0.01),其中在孕妇外周血中分别为3.73±1.03、32.53±4.36、246.50±97.30和1.47±0.14及在脐血中分别为1.30±0.45、8.03±1.24、155.00±83.50和0.47±0.46。结论:脐血中的免疫细胞幼稚,未分化成熟,免疫细胞活性及其分泌的细胞因子水平均较低,可作为一种有效、经济、安全的造血干细胞来源。

Objective： The level of CD25 and CD25 mRNA in peripheral blood mononuclear cells （PBMC）and cord blood mononuclear cells （CBMC）were explored to study the role of CD25 in expression of naive T cells. Methods：PBMC and CBMC of 30 healthy parturients were extracted from their peripheral blood and cord blood respectively with the method of Ficoll - Hypaque. The expression of CD25 before and after induction with PHA was detected by biotin - streptavidin（BSA）. Content of sIL -2R in serum were detected by EI,ISA. And CD25 mRNA was detected by real -time PCR. Results：The level of CD25 （%）in silence and after inducement, sIL- 2R （pg/ml） and CD25 mRNA（logcDNA/logGAPDH） were significantly different in periph- eral blood and in cord blood （ P 〈 0. 01 ）. And they were （3.73 ± l. 03 ）, （32.53 ± 4. 36）, （246.50 ± 97.30） and （ 1.47 ± 0.14） in peripheral blood, （1.30 ±0.45）, （8.03 ± 1.24）, （155.00 ±83.50） and （0.47 ±0.46） in cord blood, respectively. Conclusion：hnmune cells are naive and immature in cord blood and their immunocompetence and level of cytokine secretion are both low. It could be used as an effective, cheap and safe source of hematopoietic stem cells.