Early detection of postoperative sepsis and organ dysfunction using cDNA micro-array and therapeutic strategy for these complications by anti-TLR antibody.

使用 cDNA 微阵列早期检测术后败血症和器官功能障碍，并通过抗 TLR 抗体治疗这些并发症。

• 批准号: 17591374
• 负责人: KIMURA Fumio
• 金额: $ 2.05万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591374/
• 关键词: organ dysfunction; postoperative sepsis; cDNA micro-array; anti-TLR antibody; GATA3; T-bet; sC30

Background/Objectives : Aggressive hepatobiliary pancreatic surgery has been associated with high complication rates. Early detection of postoperative sepsis and organ dysfunction using cDNA micro-array and transcription-polymerase chain reaction (RT-PCR) was evaluated in patients undergoing hepatobiliary pancreatic surgery. Furthermore, therapeutic effects of anti-TLR2, 4 antibodies on these complications were determined in animal models with peritoneal sepsis or obstructive jaundice.Methods : Plasma concentrations of interleukin (IL)-6, IL-10, soluble lymphocyte activation gene (sLAG) -3, and soluble CD30 (sCD30) were determined by ELISA perioperatively in consecutive 56 patients who underwent hepatobiliary pancreatic surgery. Gene expression in PBMCs was analysed by cDNA micro-array, and expression levels of T-bet, GATA-3, TLR2, and TLR4 mRNA in PBMCs were confirmed by reverse RT-PCR.Effects of anti-TLR2, 4 antibodies on sepsis were determined in animal models with peritoneal sepsis … More or obstructive jaundice.Results : Of the 56 patients, 30 patients had postoperative infection. Postoperative plasma levels of IL-6 and IL-10 were significantly higher in patients with postoperative infection than in those without infection (p<0.05). sCD30 levels were significantly higher preoperatively, and remained higher by postoperative day 7 in patients with postoperative infection (p<0.05). sLAG-3 levels were not significantly different between two groups. GATA-3 mRNA expression level was significantly higher preoperatively, and remained higher by postoperative day 7 in patients with postoperative infection (p<0.05). T-bet mRNA expression level was significantly higher on postoperative days 3, 7, and 14 in patients with postoperative infection (p<0.05). TLR2 and TLR4 mRNA expression levels were significantly higher on postoperative day 3 in patients with postoperative infection (p< 0.05). Preoperative expression levels of GATA-3 mRNA correlated significantly with those of TLR2 and TLR4 mRNA (p<0.05). Anti-TLR2, 4 antibodies were not effective in animal models with peritoneal sepsis or obstructive jaundice.Conclusions : These results suggest that preoperative Th1/Th2 balance shifts toward Th2 dominance in patients with postoperative infection, and preoperative expression levels of circulating sCD30 and GATA3 mRNA in PBMCs are the novel markers predicting postoperative sepsis and organ dysfunction. Less

背景/目的：使用 cDNA 微阵列和转录聚合酶链反应 (RT-PCR) 对接受肝胆胰手术的患者进行评估，发现激进的肝胆胰手术与术后败血症和器官功能障碍的早期检测相关。在患有腹膜脓毒症或梗阻性黄疸的动物模型中确定了抗 TLR2,4 抗体对这些并发症的治疗效果。方法：连续 56 例接受肝胆胰手术的患者围手术期采用 ELISA 法测定血浆白细胞介素 (IL)-6、IL-10、可溶性淋巴细胞激活基因 (sLAG) -3 和可溶性 CD30 (sCD30) 的血浆浓度。通过 cDNA 微阵列分析，以及 T-bet、GATA-3、TLR2 和 TLR4 mRNA 的表达水平通过反向 RT-PCR 证实了 PBMC。在腹膜脓毒症或梗阻性黄疸动物模型中测定了抗 TLR2、4 抗体对脓毒症的影响。结果：56 名患者中，30 名患者术后血浆水平为术后感染患者的IL-6和IL-10显着高于未感染患者（p<0.05）。术后感染患者的 sLAG-3 水平在术前和术后第 7 天均保持较高水平（p<0.05）。术后感染患者中 T-bet mRNA 表达水平在术后第 3、7 和 14 天显着升高 (p<0.05)。术后第 3 天，术后感染患者的 TLR4 mRNA 表达水平显着升高 (p<0.05)，术前 GATA-3 mRNA 的表达水平与 TLR2 和 TLR4 mRNA 的表达水平显着相关 (p<0.05)。抗体在腹膜败血症或梗阻性黄疸的动物模型中无效。结论：这些结果表明，术后患者术前 Th1/Th2 平衡转向 Th2 占主导地位。感染、术前 PBMC 中循环 sCD30 和 GATA3 mRNA 的表达水平是预测术后脓毒症和器官功能障碍的新标志物。