Proteomics analysis of plasma protein complex regulating leukocyte interaction.

调节白细胞相互作用的血浆蛋白复合物的蛋白质组学分析。

• 批准号: 15590228
• 负责人: NISHIBORI Masahiro
• 金额: $ 2.3万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590228/
• 关键词: leukocyte; adhesion molecule; interaction; plasma protein; complex; cytokine; sepsis; polymyxin

Sepsis and septic shock, often associated with multiple organ failure, still remain important causes of morbidity and mortality in intensive care units. Many types of therapeutic trials for the treatment of septic shock have failed. Lipopolysaccharide(LPS) is one of the major causes of septic shock. The polymyxin B-immobilized filter column (PMX) was developed for the adsorption of endotoxin by direct hemoperfusion and has been used for the treatment of LPS-induced septic shock. In this study, we demonstrated that PMX also specifically bound monocytes from the peripheral blood leukocytes of septic patients by the analysis of bound cells using immunocytochemical and electron microscopic techniques. The specific removal of monocytes from septic patients may produce beneficial effects by reducing the interaction between activated monocytes and functionally associated cells including vascular endothelial cells. We also investigated the humoral factors adsorbed on the PMX and identified the … More some of the proteins by amino acid sequencing of the purified proteins and Western blotting. These included cytokine-like proteins and macrophage migration inhibitory factor. PMX may exert beneficial effects through the adsorption of plural factors, LPS, monocytes and humoral factors in the treatment of septic shock. We also report the plural mechanisms for controlling the activity of monocytes by the regulation of the expression of adhesion molecules.Sepsis and septic shock, often associated with multiple organ failure, still remain important causes of morbidity and mortality in intensive care units. Many types of therapeutic trials for the treatment of septic shock have failed, however, recent phase III studies using recombinant activated protein C demonstrated the effectiveness of this therapy.^<1,2> Lipopolysaccharide(LPS), one of the major causes of septic shock, together with LPS binding protein binds to CD14 on the surface of monocytes/macrophages, leading to the activation of signaling molecule complex of Toll-like receptor-4 (TLR-4) and MD2. Polymyxin B can bind LPS and neutralize its biological activity, therefore, the polymyxin B-immobilized filter (PMX) column was developed for the adsorption of endotoxin by hemoperfusion.^3 There is now increasing evidence supporting the usefulness of this treatment, showing improvement of survival rate in LPS-induced circulatory disorders and systemic inflammatory response syndrome. Moreover, the effectiveness of hemoperfusion with this column for septic shock beyond LPS endotoxemia^4 prompted us to investigate additional mechanisms. Since it is well known that different populations of leukocytes are activated during septic shock and change their adhesive phenotype, we hypothesized that some population of leukocytes may be adsorbed in the column and removed from the blood circulation after treatment. To examine this hypothesis, we investigated the cellular components in the PMX columns after direct hemofiltration in four septic patients. We also investigated the humoral factors adsorbed on the PMX. For this purpose, the proteins attached to the filter were extracted and solubilized in PBS. Some of the proteins were purified chromatograpfically and their amino acid sequences were analyzed. Less

脓毒症和脓毒性休克，往往与多器官衰竭，仍然是重要的原因，发病率和死亡率在重症监护病房。治疗感染性休克的许多类型的治疗试验都失败了。内毒素是感染性休克的主要原因之一。固定化多粘菌素B的过滤柱（PMX）被开发用于通过直接血液灌流吸附内毒素，并已用于治疗LPS诱导的感染性休克。在这项研究中，我们表明，PMX也特异性结合单核细胞从败血症患者的外周血白细胞的结合细胞使用免疫细胞化学和电子显微镜技术的分析。从脓毒症患者中特异性去除单核细胞可通过减少活化的单核细胞与功能相关细胞（包括血管内皮细胞）之间的相互作用产生有益效果。我们还研究了吸附在PMX上的体液因子， ...更多信息 通过纯化蛋白的氨基酸测序和Western印迹法对一些蛋白进行鉴定。这些包括类精氨酸蛋白和巨噬细胞移动抑制因子。PMX可能通过吸附多种因子、LPS、单核细胞和体液因子而发挥治疗感染性休克的作用。我们还报告了多种机制控制单核细胞的活性，通过调节粘附分子的表达。脓毒症和脓毒性休克，往往与多器官功能衰竭，仍然是重要的发病率和死亡率在重症监护病房的原因。许多类型的治疗感染性休克的治疗试验失败了，然而，最近的III期研究使用重组活化蛋白C证明了这种疗法的有效性。脂多糖（LPS）是脓毒性休克的主要原因之一，其与LPS结合蛋白一起结合单核细胞/巨噬细胞表面上的CD 14，导致Toll样受体-4（TLR-4）和MD 2的信号分子复合物的活化。多粘菌素B可结合LPS并中和其生物活性，因此，研制了多粘菌素B固定化滤膜（PMX），用于血液灌流中内毒素的吸附。3现在有越来越多的证据支持这种治疗的有效性，显示LPS诱导的循环障碍和全身炎症反应综合征的生存率提高。此外，使用该柱进行血液灌流治疗感染性休克的有效性超过了LPS内毒素血症^[4]，这促使我们研究其他机制。由于众所周知，不同的白细胞群体在感染性休克期间被激活并改变其粘附表型，因此我们假设一些白细胞群体可能被吸附在柱中并在治疗后从血液循环中去除。为了验证这一假设，我们研究了4例脓毒症患者直接血液滤过后PMX柱中的细胞成分。我们还研究了PMX上吸附的体液因子。为此目的，提取附着在过滤器上的蛋白质并溶解在PBS中。部分蛋白质经层析纯化并进行氨基酸序列分析。少

项目成果

Differential effect of LFA703, pravastatin, and fluvastatin on production of IL-18 and expression of ICAM-1 and CD40 in human monocytes.

LFA703、普伐他汀和氟伐他汀对人单核细胞中 IL-18 的产生以及 ICAM-1 和 CD40 表达的不同影响。

• 发表时间: 2005
• 期刊: Journal of Leukocyte Biology 77(3)
• 作者: Takahashi HK

Differential effect of PGE1 and PGE2 on LPS-induced adhesion molecules expression on monocytes.

PGE1 和 PGE2 对 LPS 诱导的单核细胞粘附分子表达的不同影响。

• 期刊: Eur J.Pharmacol. (In press.)
• 作者: Takahashi HK;Takahashi HK;Yokoyama M;Takahashi HK;Takahashi HK;Takahashi HK;Mori S;Kubo S;Tamura R;Yokoyama et al.;Takahashi et al.;Takahashi HK;Takahashi HK;Jikuhara A;Takahashi HK;Nishibori M;Mori S;Morichika T;Mori S;Sendo T;Nishibori M;Mori S;Takahashi KH;Morichika T;Takahashi HK;Takahashi et al.;Takahashi et al.;Takahashi et al.
• 通讯作者: Takahashi et al.

α1-Adrenergic receptor antagonists induce production of IL-18 and expressi of ICAM-1 and CD40 in human monocytes.

α1-肾上腺素能受体拮抗剂诱导人单核细胞产生 IL-18 以及 ICAM-1 和 CD40 的表达。

• 期刊: J.Immunother. (In press.)
• 作者: Takahashi HK;Takahashi HK;Yokoyama M;Takahashi HK;Takahashi HK;Takahashi HK;Mori S;Kubo S;Tamura R;Yokoyama et al.;Takahashi et al.;Takahashi HK;Takahashi HK;Jikuhara A;Takahashi HK;Nishibori M;Mori S;Morichika T;Mori S;Sendo T;Nishibori M;Mori S;Takahashi KH;Morichika T;Takahashi HK;Takahashi et al.;Takahashi et al.;Takahashi et al.;Takahashi et al.
• 通讯作者: Takahashi et al.

Mori et al.: "Histidine-rich glycoprotein plus zinc reverses growth inhibition of vascular smooth muscle cells by heparin"Cell Tissues Res. 312. 353-359 (2003)

Mori 等人：“富含组氨酸的糖蛋白加锌可逆转肝素对血管平滑肌细胞的生长抑制”Cell Tissues Res。

The regulation of ICAM-1 and LFA-1 interaction by autacoids and statins : a novel strategy for controlling inflammation and immune responses.

自体激素和他汀类药物对 ICAM-1 和 LFA-1 相互作用的调节：控制炎症和免疫反应的新策略。

• 发表时间: 2003
• 期刊: Journal of Pharmacological Sciences 92(1)
• 作者: Takahashi HK;Takahashi HK;Yokoyama M;Takahashi HK;Takahashi HK;Takahashi HK;Mori S;Kubo S;Tamura R;Yokoyama et al.;Takahashi et al.;Takahashi HK;Takahashi HK;Jikuhara A;Takahashi HK;Nishibori M
• 通讯作者: Nishibori M