Cole A M, Dewan P, Ganz T
Departments of Medicine and Pathology and the Will Rogers Institute for Pulmonary Research, UCLA School of Medicine, Los Angeles, California 90095, USA.
Infect Immun. 1999 Jul;67(7):3267-75. doi: 10.1128/IAI.67.7.3267-3275.1999.
Minimally manipulated nasal secretions, an accessible form of airway surface fluid, were tested against indigenous and added bacteria by using CFU assays. Antimicrobial activity was found to vary between donors and with different target bacteria and was markedly diminished by dilution of the airway secretions. Donor-to-donor differences in electrophoresis patterns of nasal secretions in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE) and acid urea-PAGE analyses were readily observed, suggesting that polymorphic genes encode the secreted proteins. Three donors (of twenty-four total), whose nasal fluid yielded similar protein band patterns and did not kill indigenous bacteria, were determined to be heavy nasal carriers of Staphylococcus aureus. Their fluid was deficient in microbicidal activity toward a colonizing strain of S. aureus but the defect was corrected in vitro by a 1:1 addition of nasal fluid from noncarriers. The microbicidal activity of normal fluid was inactivated by heating it for 10 min to 100 degrees C and could not be restored solely by the addition of two major nasal antimicrobial proteins, lysozyme and lactoferrin. Several other known antimicrobial proteins and peptides, including statherin, secretory phospholipase A2, and defensins, were identified in nasal secretions and likely contribute to their total antimicrobial properties. Nasal fluid may serve as a useful model for the analysis of lower-airway secretions and their role in host defense against airway colonization and pulmonary infections.
采用菌落形成单位(CFU)测定法,对经过最少处理的鼻腔分泌物(一种易于获取的气道表面液体形式)针对天然存在的和添加的细菌进行了检测。结果发现,抗菌活性在不同供体之间以及针对不同目标细菌时存在差异,并且气道分泌物稀释后抗菌活性会显著降低。在十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(PAGE)和酸性尿素 - PAGE分析中,很容易观察到鼻腔分泌物电泳图谱在供体之间的差异,这表明多态性基因编码分泌蛋白。在总共24名供体中,有3名供体的鼻腔分泌物产生相似的蛋白条带模式且不能杀灭天然存在的细菌,经测定他们是金黄色葡萄球菌的重度鼻腔携带者。他们的分泌物对金黄色葡萄球菌的定殖菌株缺乏杀菌活性,但通过加入1:1的非携带者鼻腔分泌物,在体外可纠正这一缺陷。正常分泌物的杀菌活性在100℃加热10分钟后失活,且仅添加两种主要的鼻腔抗菌蛋白溶菌酶和乳铁蛋白无法恢复其活性。在鼻腔分泌物中还鉴定出了其他几种已知的抗菌蛋白和肽,包括磷蛋白、分泌型磷脂酶A2和防御素,它们可能共同构成了分泌物的整体抗菌特性。鼻腔分泌物可能是分析下呼吸道分泌物及其在宿主抵御气道定殖和肺部感染中作用的有用模型。