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人中性粒细胞胞质S-100蛋白复合物(钙卫蛋白)对嗜沫二氧化碳嗜纤维菌的体外抗菌活性。

In vitro antimicrobial activity of the human neutrophil cytosolic S-100 protein complex, calprotectin, against Capnocytophaga sputigena.

作者信息

Miyasaki K T, Bodeau A L, Murthy A R, Lehrer R I

机构信息

Section of Oral Biology, UCLA School of Dentistry 90024-1668.

出版信息

J Dent Res. 1993 Feb;72(2):517-23. doi: 10.1177/00220345930720020801.

Abstract

Calprotectin is a complex of two anionic proteins found in abundance in the cytosol of neutrophils, certain macrophages, and oral epithelial keratinocytes. Bacteria of the genus Capnocytophaga are pathogens of periodontal origin which can cause systemic infection in neutropenic subjects. Recently, it has been observed that Capnocytophaga may be internalized by neutrophils within the cytosol rather than within a membrane-delimited phagosome. The purpose of this study was to test the in vitro antibacterial effect of the cytosolic complex, calprotectin, against Capnocytophaga sputigena. Calprotectin was purified from the cytosol of human neutrophils by gel filtration and anion exchange FPLC, and it exerted potent in vitro antimicrobial effects against C. sputigena. Net bacteriostatic activity was exerted up to 18 h, after which bactericidal effects were observed. Both net bacteriostatic and bactericidal activity occurred at concentrations above 20 micrograms/mL and exhibited identical dose-response characteristics. Particle counts increased in the presence of calprotectin, despite net bacteriostasis as assessed by changes in colony-forming units (CFU). Dose-response characteristics and direct particle counts suggested that net bacteriostatic effects were the result of balanced cell division and death, rather than suspension of cell division. We conclude that calprotectin can be a significant contributor to host defense against infection by Capnocytophaga.

摘要

钙卫蛋白是一种由两种阴离子蛋白组成的复合物,在中性粒细胞、某些巨噬细胞和口腔上皮角质形成细胞的胞质溶胶中大量存在。二氧化碳嗜纤维菌属细菌是牙周源性病原体,可在中性粒细胞减少的患者中引起全身感染。最近,人们观察到二氧化碳嗜纤维菌可能被中性粒细胞内化到胞质溶胶中,而不是被包被在膜限定的吞噬体中。本研究的目的是测试胞质复合物钙卫蛋白对牙龈二氧化碳嗜纤维菌的体外抗菌作用。通过凝胶过滤和阴离子交换快速蛋白质液相色谱法从人中性粒细胞的胞质溶胶中纯化钙卫蛋白,它对牙龈二氧化碳嗜纤维菌具有强大的体外抗菌作用。净抑菌活性可持续18小时,之后观察到杀菌作用。净抑菌和杀菌活性均在浓度高于20微克/毫升时出现,并表现出相同的剂量反应特征。尽管通过菌落形成单位(CFU)的变化评估为净抑菌,但在钙卫蛋白存在的情况下颗粒计数增加。剂量反应特征和直接颗粒计数表明,净抑菌作用是细胞分裂和死亡平衡的结果,而不是细胞分裂的暂停。我们得出结论,钙卫蛋白可能是宿主抵御二氧化碳嗜纤维菌感染的重要因素。

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