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，我们使用皮肤吸水器重新建立了人类供体中无菌炎症性渗出液的模型。在过去的六个月中，我们已经招募了9个捐助者，并建立了基线细胞因子反应，部分是从该实验室的先前结果复制的，但也鉴定出在泡沫形成期间未经报道的其他细胞因子上调的其他细胞因子。我们正计划使用这种水泡技术来探索免疫功能障碍的患者的炎症反应，但也通过免疫调节剂局部治疗（在泡沫部位）进行了无数治疗的患者。我们已经与渗出液细胞和外周细胞共同收集了RNA，并将使用微阵列技术来识别二氨基肌诱导的基因组调节。（Kol Zarmerm）。 我们继续研究纤维蛋白原作为IL-8产生的调节剂。在先前的工作中，我们表明，纤维蛋白原扩增在其他趋化因子（例如FMET-LEU-PHE和LTB4）刺激的中性粒细胞中的IL-8合成。我们将这项研究扩展到人的单核细胞，并表明通过单核细胞以及增加IL-6和TNFα产生的生理浓度增大IL-8的生理浓度。相比之下，纤维蛋白原对单核细胞趋化蛋白-1（MCP-1），Inteferon-beta或干扰素诱导蛋白10（IP-10）没有影响。与仅纤维蛋白原治疗相比，用纤维蛋白原（小于2 mg/ml）和补体5片段的单核细胞和补体5片段的治疗导致IL-8和IL-6产生的100％增加。这与单核细胞IL-8 mRNA和NF-KB活性的短暂增加有关。与正常单核细胞相比，通过TOLL样受体途径（NEMO缺乏症和IRAK-4缺乏症）的单核细胞通过TOLL样受体途径（NEMO缺乏和IRAK-4缺乏症）的单核细胞降低了IL-8对纤维蛋白原的反应80％。此外，正常的单核细胞对纤维蛋白原的反应被阻断CD14的抗体阻断，CD14是LPS受体的亚基，该亚基传递信号TLR TLR4。MY4对PMA和IONYMYCIN诱导的细胞因子产生没有影响。 （面团库恩斯） 今年，我们利用患者患有类似收费的受体途径和复发性细菌感染（IRAK-4缺乏症）来研究嗜中性粒细胞启动，以通过内毒素产生超氧化物。我们已经表明，IRAK-4缺乏症已响应内毒素，并减少了胞质NADPH氧化酶蛋白p47phox和p67phox对质膜的脱氧启动。通过使用IMGenix＃img-417e制造的TLR-4阻断抗体，通过阻止正常细胞中的Toll样受体4（TLR-4）信号传导获得类似的结果。这些研究表明，TLR途径在NADPH氧化酶途径的内毒素启动中起着重要作用。 （Anjali Singh）