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目的:分析G-CSF动员前后外周血T细胞中CD3分子的γ,δ,ε和ζ链基因表达水平变化特点,以深入了解G-CSF对T细胞信号转导的影响。方法:利用SYBR GreenⅠ实时荧光定量RT-PCR检测17例健康供者G-CSF动员前后外周血单个核细胞中CD3γ、δ、ε和ζ链的表达水平,以β2微球蛋白基因(β2MG)作为内参照,采用相对定量公式:2-ΔCt×100%,计算CD3分子各亚单位的相对mRNA表达量。结果:G-CSF动员前后4种CD3基因表达水平模式均为CD3ζ>CD3ε>CD3δ>CD3γ。供者G-CSF动员后CD3ζ和ε链基因表达水平(2.194%,0.867%)明显低于G-CSF动员前(7.615%,1.839%),动员前后比较存在显著差异(P=0.031,P=0.009);动员前CD3δ和γ基因平均表达水平分别为1.097%和0.271%,与动员后比较(0.333%和0.077%)并没有显著性差异(P=0.093,P=0.070)。G-CSF动员前与动员后CD3ζ基因表达水平存在正相关性(rs=0.576,P=0.016);G-CSF动员前与动员后CD3γ、δ和ε基因表达水平均不存在显著相关性(P=0.726,P=0.236,P=0.198)。G-CSF动员前与动员后CD3ζ基因以及G-CSF动员后CD3δ基因表达水平与供者年龄均有负相关性(rs=-0.549,P=0.023;rs=-0.679,P=0.003;rs=-0.570,P=0.017)。结论:本研究结果提示G-CSF动员可能主要影响TCR信号通路中的CD3ζ和ε链基因的表达,而整体的CD3基因表达模式未发生改变。
OBJECTIVE: To analyze the changes of γ, δ, ε and ζ chain gene expression of CD3 in peripheral blood T lymphocytes before and after G-CSF mobilization to understand the effect of G-CSF on T cell signal transduction. Methods: The expression levels of CD3γ, δ, ε and ζ chains in peripheral blood mononuclear cells of 17 healthy donors before and after G-CSF mobilization were detected by SYBR Green Ⅰ real-time fluorescent quantitative RT-PCR. Β2 microglobulin gene (β2MG) Internal reference, the relative quantification formula: 2-ΔCt × 100%, calculate the CD3 molecular subunit relative mRNA expression levels. Results: The expression patterns of four CD3 genes before and after G-CSF mobilization were all CD3ζ> CD3ε> CD3δ> CD3γ. The gene expression levels of CD3ζ and ε chain after donor G-CSF mobilization (2.194% and 0.867%) were significantly lower than those before G-CSF mobilization (7.615% and 1.839%, P = 0.031, P = 0.009). The mean CD3δ and γ gene expression levels before mobilization were 1.097% and 0.271%, respectively, which were not significantly different from those after mobilization (0.333% and 0.077%, P = 0.093, P = 0.070). There was a positive correlation between G-CSF mobilization and CD3ζ gene expression (rs = 0.576, P = 0.016). There was no significant correlation between G-CSF mobilization and CD3γ, δ and ε gene expression = 0.726, P = 0.236, P = 0.198). There was a negative correlation between CD3δgene expression level and donor age before and after mobilization of CD3ζ gene and G-CSF mobilization (rs = -0.549, P = 0.023; rs = -0.679, P = 0.003; rs = -0.570, P = 0.017). Conclusion: Our results suggest that mobilization of G-CSF may mainly affect the expression of CD3ζ and ε chain genes in the TCR signaling pathway, while the overall CD3 gene expression pattern has not changed.