Quantitative assay of cytokine mRNA expression in peripheral blood mononuclear cells and its clinical application

以外周血单个核细胞（PBM）细胞因子mRNA为内对照，采用定量RT-PCR方法，建立了外周血单个核细胞（PBM）细胞因子mRNA表达的定量检测方法。利用该检测系统，定量评估il - 2激活后PBM中il - 1- α、il - 1- β或tnf - α的mRNA表达。il - α和- β mRNA的表达在激活后1 ~ 3小时和6 ~ 12小时呈双相增加，随后呈下降趋势。然而，tnf - α mRNA的表达在刺激24至48小时后逐渐增加，峰值水平为1至3小时。此外，在体内用il - 2治疗的癌症患者中，我们评估了PBM中这些细胞因子mRNA的表达。治疗后1天观察到il - α或- β mRNA表达增加，而tnf - α mRNA在治疗后差异表达。这些结果表明，该检测系统可用于研究各种BRM对癌症患者细胞因子网络的影响，从而评估这些BRM对癌症的有效性。

A quantitative assay for the expression of cytokine mRNA in peripheral blood mononuclear cells (PBM) has been developed using a quantitative RT-PCR method, in which cRNA of each cytokine is employed as inner control. Utilizing this assay system, mRNA expression of IL1-alpha、IL1-beta or TNF-alpha was quantitatively assessed in PBM after activation with IL2 in vitro. Both IL1-alpha and -beta mRNA expression was biphasically increased by 1 to 3 hr and 6 to 12 hr after activation, and thereafter decreased. However, expression of TNF-alpha mRNA was gradually increased after 24 to 48 hr stimulation following the peak level of evelation by 1 to 3 hr. Further, these cytokine mRNA expressions in PBM were evaluated in patients with carcinoma following the treatment with IL2 in vivo. The increased expression of IL1-alpha or -beta mRNA was observed 1 day after the treatment, while TNF-alpha mRNA was differentially expressed following the therapy.These results indicate that this assay system could be useful for the investigation of the influence of various BRM on cytokine-network in cancer patients and, therefore, for the evaluation of the effectiveness of these BRM on cancer.