DEFENSINS - NATURAL PEPTIDE ANTIBIOTICS OF HUMAN-NEUTROPHILS
DEFENSINS - NATURAL PEPTIDE ANTIBIOTICS OF HUMAN-NEUTROPHILS
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DOI：
10.1172/jci112120
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发表时间：
1985-01-01
影响因子：
15.9
通讯作者：
LEHRER, RI
中科院分区：
文献类型：
--
作者：
GANZ, T;SELSTED, ME;LEHRER, RI
我们从正常人中性粒细胞中提取了富含颗粒的沉淀物，并对其进行了层析、电泳和功能分析。该提取物含有三个小的(相对分子质量为3500)的抗菌肽，分别命名为人中性粒细胞多肽(HNP)-1、HNP-2和HNP-3，将其称为防御素。这三种防御素的混合物HNP 1-3在含有一定营养的10 mM磷酸盐缓冲液中对金黄色葡萄球菌、铜绿假单胞菌和大肠埃希菌有较好的体外杀灭作用，但在无营养缓冲液中几乎没有杀菌活性。相比之下，无营养缓冲液支持HNP 1-3对新生隐球菌的高度活性。除了其抗菌和抗真菌特性外，HNP 1-3直接灭活了1型单纯疱疹病毒。两种单独纯化的防御素，HNP-1和HNP-2，与HNP 1-3的混合物一样具有杀菌作用，HNP-3对大多数但不是所有被测试的微生物的活性低于其他防御素。免疫过氧化物酶染色显示HNP 1-3在光学显微镜下呈颗粒状定位于中性粒细胞的S细胞质中。冰冻切片免疫金透射电子显微镜显示HNP 1-3定位于嗜天青颗粒中。这些研究定义了人中性粒细胞的广谱抗菌系统。防御素系统可能与氧气依赖的杀菌过程一起工作，或独立于氧依赖的杀菌过程运行，使人中性粒细胞灭活和破坏潜在的病原体。
We extracted a granule-rich sediment from normal human neutrophils and subjected it to chromatographic, electrophoretic, and functional analysis. The extract contained three small (molecular weight < 3500) antibiotic peptides that were named human neutrophil peptide (HNP)-1, HNP-2, and HNP-3, and which will be referred to as "defensins". HNP 1-3, a mixture of the three defensins, killed Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli effectively in vitro when tested in 10 mM phosphate buffer containing certain nutrients, but it had little or no bactericidal activity in nutrient-free buffer. In contrast, the nutrient-free buffer supported a high degree of activity by HNP 1-3 against Cryptococcus neoformans. In addition to its antibacterial and antifungal properties, HNP 1-3 directly inactivated herpes simples virus, Type 1. Two of the individual purified defensins, HNP-1 and HNP-2, were as microbicidal as the mixture HNP 1-3, HNP-3 was less active than the other defensins against most but not all of the microbes tested. Immunoperoxidase stains revealed HNP 1-3 to have a granular localization in the neutrophil''s cytoplasm by light microscopy. Frozen thin section immunogold transmission electron microscopy showed HNP 1-3 to be localized in azurophil granules. These studies define a broad-spectrum antimicrobial system in human neutrophils. The defensin system may operate in conjunction with or independently from oxygen-dependent microbicidal processes to enable human neutrophils to inactivate and destroy potential pathogens.