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)中各细胞因子cRNA为内对照，建立了定量RT-PCR检测PBM中细胞因子mRNA表达的方法。利用该检测系统，定量检测IL-2体外激活后外周血中IL-1-α、IL-1-β和、-α的表达。IL-1-α和-βmRNA的表达在激活后1~3小时和6~12小时呈双相性升高，此后下降。而在24~48小时刺激后1~3小时，肿瘤坏死因子-αmRNA的表达逐渐增加。此外，在体内用IL2治疗后的癌症患者的PBM中评估了这些细胞因子mRNA的表达。IL-1-α或-βmRNA在治疗后1天开始升高，而肿瘤坏死因子-α在治疗后有不同程度的表达，提示该检测体系可用于研究不同BRM对肿瘤患者细胞因子网络的影响，从而评价这些BRM对癌症的疗效。

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.