Isolation and characterization of human β-defensin-3, a novel human inducible peptide antibiotic
Isolation and characterization of human β-defensin-3, a novel human inducible peptide antibiotic
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DOI：
10.1074/jbc.m008557200
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发表时间：
2001-02-23
影响因子：
4.8
通讯作者：
Schröder, JM
中科院分区：
文献类型：
--
作者：
Harder, J;Bartels, J;Schröder, JM
由多重耐药革兰氏阳性菌（特别是金黄色葡萄球菌）引起的感染引起的公共卫生问题日益严重，促使我们筛查人类上皮细胞中的内源性金黄色葡萄球菌杀灭因子。一种新型 5-kDa、非溶血性抗菌肽（人 β-防御素-3，hBD-3）是从人类皮损银屑病鳞屑中分离出来并从角质形成细胞中克隆出来的，hBD-3 表现出对盐不敏感的广谱有效抗菌活性，可对抗许多潜在的致病微生物，包括多重耐药金黄色葡萄球菌和 耐万古霉素屎肠球菌。 hBD-3 处理的金黄色葡萄球菌的超微结构分析显示细胞壁穿孔的迹象。重组 hBD-3（在大肠杆菌中表达为 His-Tag 融合蛋白）和化学合成的 hBD-3 在抗菌活性和生化特性方面与天然存在的肽没有区别。对不同组织的研究表明皮肤和扁桃体是主要的 hBD-3 mRNA 表达组织。对细胞培养上清液中抗菌肽的分子克隆和生化分析表明角质形成细胞和气道上皮细胞是 hBD-3 的细胞来源。发现肿瘤坏死因子 α 和与细菌接触可诱导 hBD-3 mRNA 表达。因此，hBD-3 可能在皮肤和肺部各种微生物感染（例如囊性纤维化）的先天上皮防御中发挥重要作用。
The growing public health problem of infections caused by multiresistant Gram-positive bacteria, in particular Staphylococcus aureus, prompted us to screen human epithelia for endogenous S, aureus-killing factors. A novel 5-kDa, nonhemolytic antimicrobial peptide (human beta -defensin-3, hBD-3) was isolated from human lesional psoriatic scales and cloned from keratinocytes, hBD-3 demonstrated a salt-insensitive broad spectrum of potent antimicrobial activity against many potentially pathogenic microbes including multiresistant S. aureus and vancomycin-resistant Enterococcus faecium. Ultrastructural analyses of hBD-3-treated S, aureus revealed signs of cell wall perforation. Recombinant hBD-3 (expressed as a His-Tag-fusion protein in Escherichia coli) and chemically synthesized hBD-3 were indistinguishable from naturally occurring peptide with respect to their antimicrobial activity and biochemical properties. Investigation of different tissues revealed skin and tonsils to be major hBD-3 mRNA-expressing tissues. Molecular cloning and biochemical analyses of antimicrobial peptides in cell culture supernatants revealed keratinocytes and airway epithelial cells as cellular sources of hBD-3. Tumor necrosis factor alpha and contact with bacteria were found to induce hBD-3 mRNA expression. hBD-3 therefore might be important in the innate epithelial defense of infections by various microorganisms seen in skin and lung, such as cystic fibrosis.