Effect Of Cytokines In Host Defense And Inflammation

细胞因子在宿主防御和炎症中的作用

• 批准号: 7299946
• 负责人: JOHN I GALLIN
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7299946
• 关键词: Effect; Cytokines; Host; Defense; Inflammation

This year we reestablished a model of sterile inflammatory exudates in human donors using skin suction blisters. Over the past six months we have enrolled 9 donors and have established a baseline cytokine response in part reproducting previous results from this lab, but also identifying additional cytokines not previously reported to be upregulated during blister formation. We are planning on using this blister technique to explore inflammatory responses in patients with immune dysfunction but also innormal patients treated topically (at the blister site) with immunomodulatoryagents. We have collected RNA concurently with exudate cells and from peripheral cells and will be using microarray technology to identify genomic regulation induced by diapediesis.(Kol Zarember) We continued our studies of fibrinogen as a regulator of IL-8 production. In previous work we showed that fibrinogen amplifies IL-8 synthesis in neutrophils stimulated with other chemoattractants such as fmet-leu-phe and LTB4. We extended this studies to human monocytes and showed that addition of physiological concentrations of fibrinogen amplified IL-8 production by monocytes as well as increased IL-6 and TNF alpha production. In contrast, fibrinogen had no effect on monocyte chemoattractant protein-1 (MCP-1), inteferon-beta, or interferon inducible protein-10 (IP-10). Treatment of monocytes with fibrinogen (less than 2 mg/ml) and complement 5 fragment, C5a, resulted in a 100% increase in both IL-8 and IL-6 production, compared to fibrinogen treatment alone. This was associated with a transient increase in monocyte IL-8 mRNA and NF-kB activity. Monocytes from patients with defective LPS and IL-1 signaling through the Toll-like receptor pathway (NEMO deficiency and IRAK-4 deficiency)had 80% reduced IL-8 response to fibrinogen compared with normal monocytes. Moreover, normal monocyte responses to fibrinogen were blocked by antibody that blocks CD14, a subunit of the LPS receptor that transduces signal throug TLR 4. MY4 had no effect on cytokine production induced by PMA and ionomycin. (Dough Kuhns) This year we have utilized our patient with a disorder of the Toll-like receptor pathway and recurrent bacterial infections (IRAK-4 deficiency), to study neutrophil priming for superoxide production by endotoxin. We have shown that IRAK-4 deficiency has dminished priming of superoxide production in response to endotoxin and diminished translocation of cytosolic NADPH oxidase proteins p47phox and p67phox to the plasma membrane. Similar results were obtained by blocking toll-like receptor 4(TLR-4) signalling in normal cells using the TLR-4 blocking antibody made by Imgenix #IMG-417E). These studies indicate an importrant role for the TLR pathway in endotoxin priming of NADPH oxidase pathway. (Anjali Singh)

今年，我们重新建立了一个模型，无菌炎症渗出液在人类捐赠者使用皮肤抽吸水泡。在过去的六个月里，我们招募了9名供体，并建立了基线细胞因子反应，部分重现了该实验室以前的结果，但也鉴定了以前未报告在水疱形成期间上调的其他细胞因子。我们正计划使用这种水泡技术来探讨免疫功能障碍患者的炎症反应，以及用免疫调节剂局部治疗（在水泡部位）的正常患者的炎症反应。我们同时从渗出细胞和外周细胞中收集RNA，并将使用微阵列技术来鉴定由渗出诱导的基因组调控。(Kol Zarember） 我们继续研究纤维蛋白原作为IL-8产生的调节剂。在以前的工作中，我们表明，纤维蛋白原放大IL-8的合成与其他化学引诱剂，如fmet-leu-phe和LTB 4刺激的中性粒细胞。我们将这项研究扩展到人单核细胞，并表明加入生理浓度的纤维蛋白原可扩增单核细胞的IL-8产生，并增加IL-6和TNF α的产生。相反，纤维蛋白原对单核细胞趋化蛋白-1（MCP-1）、干扰素-β或干扰素诱导蛋白-10（IP-10）没有影响。与单独的纤维蛋白原处理相比，用纤维蛋白原（小于2 mg/ml）和补体5片段C5 a处理单核细胞导致IL-8和IL-6产生增加100%。这与单核细胞IL-8 mRNA和NF-κ B活性的瞬时增加有关。与正常单核细胞相比，LPS和IL-1通过Toll样受体途径（NEMO缺陷和IRAK-4缺陷）信号传导缺陷患者的单核细胞对纤维蛋白原的IL-8反应降低了80%。此外，正常单核细胞对纤维蛋白原的反应被阻断CD 14的抗体阻断，CD 14是LPS受体的亚单位，其转导信号转导TLR 4。MY 4对PMA和离子霉素诱导的细胞因子产生无影响。（Dough Kuhns） 今年，我们利用我们的病人与Toll样受体途径的障碍和反复细菌感染（IRAK-4缺乏症），研究中性粒细胞引发的超氧化物产生的内毒素。我们已经表明，IRAK-4缺陷减少了响应内毒素的超氧化物产生的启动，并减少了胞质NADPH氧化酶蛋白p47 phox和p67 phox向质膜的转运。通过使用由Imgenix制造的TLR-4阻断抗体阻断正常细胞中的toll样受体4（TLR-4）信号传导获得了类似的结果（#IMG-417 E）。这些研究表明TLR途径在NADPH氧化酶途径的内毒素引发中起重要作用。（安贾利·辛格）