Role of Endotoxin-binding proteins in Non-specific Protection to Infection

内毒素结合蛋白在非特异性感染保护中的作用

• 批准号: 04670250
• 负责人: HIRATA Michimasa
• 金额: $ 1.34万
• 依托单位: Iwate Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 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-04670250/
• 关键词: Lipopolysaccharide(LPS); LPS-binding protein; Tissue factor; Disseminated intravascular coagulation; Anticoagulant; Antibacteria; LPS-neutralization; リポ多種(LPS); LPS中和; 酸化窒素; 腫瘍壊死因子; エンドトキシンショック

Endotoxin(lipopolysaccharide=LPS), cell wall component of gram-negative bacteria, activates monocytes and macrophages to release cytokines, reactive nitrogen intermediates (RNI), and to generate tissue factor(TF) which initiate coagulation. We have purified 7kDa and 18kDa cationic antibacterial proteins (CAP-7 and CAP-18) with LPS-binding and LPS-neutralizing activities from rabbit granulocytes using as an assay the agglutination of erythrocytes coated with Re-LPS.From protein sequencing, CAP-7 was identified as the C-terminal 37 amino acid fragment of CAP-18. Synthetic peptide #197(identical sequence to CAP-7, Gly1-Try37) and #36-1 (a truncation of CAP consisting of 32 amino acid residues, Gly1-Ala32) showed LPS-binding activity. Each peptide inhibited LPS-induced tissue factor(TF) generation by murine peritoneal macrophages, even added 1 hour after stimulation of cells with LPS.C57BL/6 mice treated with #197 were significantly protected from lethal LPS challenge. Peptide #36 also blo … More cked the LPS-induced lethality. These peptides had antibacterial activity to gram-negative bacteria, such as E.coli, S.typhimurium, K.pneumonia, Ps.aeruginosa and also to gram-positive S.aureus(Methicllin sensitive and resistant strains). Both peptides inhibited TF-and Xa-induced plasma clotting. Using synthetic chromogenic substrates, both CAP7 peptides blocked the coagulation cascade at two sites, activation of factor X to Xa and conversion of Factor II(prothrombin) to factor IIa(prothrombin). In vivo treatment of peptide #197 prevented acute lethality in mice injected with tissue factor (rabbit brain thromboplastin). Two other peptides, #32(Gly1-Phe9) and #50(lle13-Typ37) failed to demonstrate LPS-binding, LPS-neutralizing, antibacterial and anticoagulant activities. The active peptides but not the inactive peptide maintain a putative heparin binding domain at their N-termini. This heparin binding domain is participate in the LPS-binding, LPS neutralizing, antibacterial and anticoaThese active peptides may have a therapeutic potential for treatment for DIC due to sepsis and endotoxin shock. Less

内毒素（脂多糖= LPS），革兰氏阴性细菌的细胞壁成分，激活单核细胞和巨噬细胞以释放细胞因子，反应性氮中间体（RNI）以及产生启动凝结的组织因子（TF）。 We have purified 7kDa and 18kDa cationic antibacterial proteins (CAP-7 and CAP-18) with LPS-binding and LPS-neutralizing activities from rabbit granulocytes using as an assay the agglutination of erythrocytes coated with Re-LPS.From protein sequencing, CAP-7 was identified as the C-terminal 37 amino acid fragment of CAP-18.合成肽＃197（与CAP-7，Gly1-Try37相同的序列）和＃36-1（cap的截断由32个氨基酸残基组成，GLY1-ALA32）显示了LPS结合活性。每种肽抑制LPS诱导的组织因子（TF）通过鼠腹膜巨噬细胞产生，甚至在用＃197处理的LPS.C57BL/6小鼠刺激细胞后增加了1小时，也受到了致命的LPS挑战的显着保护。肽＃36也是Blo…更多地介绍了LPS引起的致命性。这些胡椒体具有革兰氏阴性细菌的抗菌活性，例如e.coli，s. typhimurium，k.pneumonia，pneumonia，ps.aeruginosa以及革兰氏阳性的s. aureus（甲基链磷脂敏感和抗性菌株）。在体内对肽＃197的体内治疗中的两种辣椒都可以防止注射组织因子（兔脑血栓蛋白）的小鼠的急性致死性。另外两个肽，＃32（Gly1-Phe9）和＃50（LLE13-TYP37）未能证明LPS结合，LPS中和，抗菌和抗凝作用。活性肽，但不能在其N末端保持推定的肝素结合结构域。该肝素结合结构域参与LPS结合，LPS中和，抗菌和抗疾病活性肽的活性肽可能具有治疗潜力，可用于治疗DIC，导致败血症和内毒素休克。较少的

项目成果

Hirata, M., Shimomura, Y., Yoshida, M., Morgan, J.G., Palings, I., Wilson D., Yen, M.H. : and Larrick, J.W.: "Characterization of a rabbit cationic protein (CAP-18) with lipopolysaccharide-inhibitory activity" Infect.Immun. (in press). (1994)

平田 M.、下村 Y.、吉田 M.、摩根 J.G.、帕林斯 I.、威尔逊 D.、日元 M.H.

Hirata M.,Shimomura Y et al: "Endotoxin-neutralizing,antibacterial and anticoagulant activities rabbit CAP18-derived peptides." Gram Negative Sepsis:Basic Science to clinical Investigation. (in press). (1994)

Hirata M.、Shimomura Y 等人：“兔 CAP18 衍生肽具有中和、抗菌和抗凝血活性。”

Larrick.J.W,Hirata,Metsl: "A novelgranalocyte-derived peptide with LPS neutralizing activity" J.Immunol. 152. 231-240 (1994)

Larrick.J.W、Hirata、Metsl：“一种具有 LPS 中和活性的新型粒细胞衍生肽”J.Immunol。

平田 陸正: "内毒素結合蛋白質の構造と活性" 第39回毒素シンポジウム予稿集. 80-84 (1992)

Rikumasa Hirata：“内毒素结合蛋白的结构和活性”第 39 届毒素研讨会论文集 80-84（1992 年）。

Larrick,J.W.,Hirata,M et al: "A Novel granulocyte-derived peptide with LPS neutralizing activity" J Immunol. 152. 231-240 (1994)

Larrick,J.W.,Hirata,M 等人：“一种具有 LPS 中和活性的新型粒细胞衍生肽”JImmunol。