Monitaring of rejection after transplantation by cytokine mRNA mapping

通过细胞因子 mRNA 图谱监测移植后排斥反应

• 批准号: 04454336
• 负责人: TSUZUKI Toshiharu
• 金额: $ 4.16万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454336/
• 关键词: cytokine; mRNA; transplantation; monitaring; rejection; 末梢血; 免疫学的モニタリング

We developed a method of detecting cytokine mRNA from unfractionated whole blood using RT-PCR.IL-1beta and TNF gene expression could be detected from a 0.5ml blood sample. Total RNA was extracted by 4M guanidinium isothiocyanate mixture, 0.2M sodium acetate, phenol and chloroform and 10mug total RNA was obtained. The mRNA is reverse transcribed, and IL-1beta, TNF and IL-2R cDNA were selectively amplified by synthetic primers with PCR.Pheripheral blood was obtained from healthy volunteers and patients with severe infectons, operations and renal transplantation/Il-1beta, TNF and IL-2R mRNA were undetectable in blood of healthy volunteers. While in patients with severe infection and operation, only IL-1beta and TNF, but not IL-2R mRNA were detected. IL-2 mRNA was detectec in blood of the patients with renal transpantation. This method avoids artificial gene activation and may be applicable to monitoring cytokine gene espression in various pathophysiological state.

我们建立了一种用RT-PCR检测普通全血细胞因子mRNA的方法，每0.5ml血标本可检测到IL-1 β和TNF基因的表达。用4 M异硫氰酸胍混合液、0.2M乙酸钠、苯酚和氯仿提取总RNA，得到10 μ g总RNA。用PCR方法选择性扩增IL-1 β、TNF和IL-2 R的cDNA，并对正常人、严重感染、手术和肾移植患者的外周血进行检测，正常人外周血中未检测到IL-1 β、TNF和IL-2 R的mRNA。而在严重感染和手术患者中，仅检测到IL-1 β和TNF，而未检测到IL-2 R mRNA。肾移植患者血液中检测到IL-2 mRNA。该方法避免了人工基因激活，可用于各种病理生理状态下细胞因子基因表达的监测。

项目成果

都築 俊治: "左外側区域切除術" 手術. 47. 433-435 (1993)

Toshiharu Tsuzuki：“左外侧肺段切除术”手术。 47. 433-435 (1993)

都築 俊治: "肝切除による肝細胞癌の治療" Medicina. 29. 1660-1662 (1992)

Toshiharu Tsuzuki：“肝切除术治疗肝细胞癌”Medicina 29. 1660-1662 (1992)。

都築 俊治: "肝切除中の下大静脈損傷とその処置" 外科診療. 34. 1301-1304 (1992)

Toshiharu Tsuzuki：“肝切除术中的下腔静脉损伤及其治疗”《外科诊所》34. 1301-1304 (1992)。

若林剛: "肝虚血・再潅流障害における炎症性サイトカインの役割" 現代医療. 25. 3911-3917 (1993)

Tsuyoshi Wakabayashi：“炎症细胞因子在肝缺血/再灌注损伤中的作用”现代医学 25. 3911-3917 (1993)。

Sugioka: "Postresection prognosis of patients with hepatocellular carcinoma" Surgery. 113. 612-618 (1993)

Sugioka：“肝细胞癌患者的术后预后”手术。