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

外周血单个核细胞细胞因子mRNA表达定量测定及其临床应用

• 批准号: 05671012
• 负责人: ARINAGA Shinya
• 金额: $ 1.41万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05671012/
• 关键词: Cytokine; Messenger RNA; Quantitative assay; Interleukin l-alpha; Interleukin l-beta; Interleukin 2; Tumor necrosis factor-alpha; Immunochemotherapy; インターロイキン6; インターリューキン2; インターリューキン6

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.

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。

项目成果

S.Arinaga et al.: "Laboratory correlates of chemoimmunotherapy with low-dose recombinant interleukin-2 and mitomycin C in patients with advanced carcinoma" Cancer Invest. 12. 588-596 (1994)

S.Arinaga 等人：“晚期癌症患者中低剂量重组白细胞介素 2 和丝裂霉素 C 化学免疫疗法的实验室相关性”Cancer Invest。

S.Arinaga,M.Adachi et al: "Cytotoxic cell function and peripheral analysis of peripheral blood monon〓〓le〓r cells in cancer patients treutacl with low-〓〓〓〓〓 interluk〓〓2 and 〓〓" Cancer Immunology Immunotherapy. 37. 220-226 (1993)

S. Arinaga、M. Adachi 等人：“使用低〓〓〓〓〓 interluk〓〓2 和〓〓的癌症患者 treutacl 的外周血单〓〓le〓细胞的细胞毒性细胞功能和外周分析”癌症免疫学免疫疗法. 37. 220-226 (1993)

Karimine,N,Arinaga.S.etal: "Lymphokine-activated killer cell function of periphenal bood mowoneuclear cells,spleeucells and vegional lymph node cells in gastric cancer pationts." Clin,Exp.Immunol.96. 484-490 (1994)

Karimine,N,Arinaga.S.等人：“胃癌患者周围血液核细胞、脾细胞和植物淋巴结细胞的淋巴因子激活的杀伤细胞功能。”

足立昌士,有永信哉他: "末梢血単核球のTNFmRNAの定量的測定と臨床応用" 日本外科学会雑誌. 94. 1124-1124 (1993)

Masashi Adachi、Shinya Arunaga 等：“外周血单核细胞中 TNF mRNA 的定量测量及其临床应用”日本外科学会杂志 94. 1124-1124 (1993)。

S.Arinaga et al.: "Cytotoxic cell function and phenotypic analysis of peripheral blood mononuclear cells in cancer patients treated with low-dose interleukin 2 and mitomycin C" Cancer Immunol Immunother. 37. 220-226 (1993)

S.Arinaga 等人：“用低剂量白细胞介素 2 和丝裂霉素 C 治疗的癌症患者外周血单核细胞的细胞毒性细胞功能和表型分析”Cancer Immunol Nutritionother。