Adachi M, Inoue H, Arinaga S, Li J, Ueo H, Mori M, Akiyoshi T
Department of Surgery, Medical Institute of Bioregulation, Kyushu University, Beppu, Japan.
Cancer Immunol Immunother. 1997 Aug;44(6):329-34. doi: 10.1007/s002620050390.
After activation with interleukin-2 (IL-2), peripheral blood mononuclear cells (PBMC) have been reported to induce the expression of mRNA coding various cytokines, including interleukin(IL)-1alpha, -1beta and tumor necrosis factor alpha (TNF alpha). We examined the cytokine mRNA expression of PBMC following treatment with IL-2 in vitro and in vivo by a quantitative method using the reverse transcription/polymerase chain reaction (RT-PCR). After stimulating PBMC with IL-2 in vitro, peak levels of IL-1alpha mRNA were reached between 3 h and 12 h, and thereafter declined. The IL-1beta expression increased, with levels peaking at 1-6 h and, had decreased by 96 h. The expression of TNF alpha was elevated both 1-3 h and 24-48 h after stimulation. The peak levels of IL-1alpha and -1beta mRNA and the early elevation of TNF alpha mRNA mainly accounted for the cytokine mRNA expression in adherent cells; however, the late induction of TNF alpha mRNA was observed in nonadherent cells. In patients with advanced carcinoma, the IL-1alpha and -1beta mRNA expression were elevated after IL-2 treatment for 5 consecutive days, while the expression of TNF alpha mRNA also increased. These results indicate that the quantitative RT-PCR method appears to be useful for analyzing the cytokine mRNA expression of PBMC after treatment with IL-2.
据报道，用白细胞介素-2（IL-2）激活外周血单个核细胞（PBMC）后，可诱导编码多种细胞因子的mRNA表达，包括白细胞介素（IL）-1α、-1β和肿瘤坏死因子α（TNFα）。我们采用逆转录/聚合酶链反应（RT-PCR）定量方法，在体外和体内检测了IL-2处理后PBMC的细胞因子mRNA表达。体外使用IL-2刺激PBMC后，IL-1α mRNA在3小时至12小时之间达到峰值水平，随后下降。IL-1β表达增加，在1至6小时达到峰值，到96小时时下降。TNFα的表达在刺激后1至3小时和24至48小时均升高。IL-1α和-1β mRNA的峰值水平以及TNFα mRNA的早期升高主要出现在贴壁细胞中的细胞因子mRNA表达中；然而，在非贴壁细胞中观察到TNFα mRNA的晚期诱导。在晚期癌症患者中，连续5天接受IL-2治疗后，IL-1α和-1β mRNA表达升高，同时TNFα mRNA表达也增加。这些结果表明，定量RT-PCR方法似乎有助于分析IL-2处理后PBMC的细胞因子mRNA表达。
