New Jersey Center for Biomaterials, Rutgers University, 145 Bevier Rd., Piscataway, NJ, 08854, USA.
Department Hematology & Hematologic Malignancies, University of Utah, Salt Lake City, USA.
J Transl Med. 2019 Mar 1;17(1):68. doi: 10.1186/s12967-019-1812-8.
Human amniotic fluid (AF) contains numerous nutrients, trophic factors and defense proteins that provide a nurturing and protective environment for fetal development. Based on reports that AF has antibacterial, anti-inflammatory and regenerative properties, we designed a novel method to process AF for use in clinical care.
Six randomly selected lots of processed AF (pAF) were examined to determine whether they retained their antibacterial activity against a panel of wound-associated pathogens E. faecium, S. aureus, K. pneumoniae, A. baumannii, P. aeruginosa, and E. aerogenes (ESKAPE). To identify proteins in pAF that might be responsible for its antibacterial activity, three different lots of pAF were analyzed with quantitative cytokine arrays that consisted of 400 unique human proteins. One protein identified by microarrays, lactoferrin, and a second prominent antibacterial protein that was not identified by microarrays, lysozyme, were examined by depletion experiments to determine their contribution to the antibacterial activity of pAF.
All six lots of pAF exhibited antibacterial activity against ESKAPE microorganisms, especially against the pathogens predominately found in chronic wounds (i.e. S. aureus and P. aeruginosa). Thirty-one of the peptides on the microarray were annotated as having antibacterial activity and 26 of these were detected in pAF. Cystatin C and lactoferrin were among the most highly expressed antibacterial proteins in pAF. Cystatin C and lactoferrin were confirmed by ELISA to be present in pAF along with lysozyme. Immunoprecipitation of lactoferrin and lysozyme reduced, but did not abolish the antibacterial activities of pAF.
Our data demonstrate that pAF maintains antibacterial activity via the preservation of antibacterial proteins against a broad spectrum of wound-associated pathogens.
人类羊水（AF）含有许多营养物质、营养因子和防御蛋白，为胎儿发育提供了滋养和保护的环境。基于羊水具有抗菌、抗炎和再生特性的报告，我们设计了一种新的方法来处理 AF 用于临床护理。
检查了随机选择的六批处理后的 AF（pAF），以确定它们是否保留了对一组与伤口相关的病原体粪肠球菌、金黄色葡萄球菌、肺炎克雷伯菌、鲍曼不动杆菌、铜绿假单胞菌和阴沟肠杆菌（ESKAPE）的抗菌活性。为了确定 pAF 中可能具有抗菌活性的蛋白质，用包含 400 种独特人类蛋白质的定量细胞因子阵列分析了三种不同批次的 pAF。通过微阵列鉴定出一种蛋白质乳铁蛋白，以及另一种未通过微阵列鉴定出的突出的抗菌蛋白溶菌酶，通过耗尽实验来确定它们对 pAF 抗菌活性的贡献。
所有六批 pAF 均对 ESKAPE 微生物表现出抗菌活性，尤其是对慢性伤口中主要发现的病原体（即金黄色葡萄球菌和铜绿假单胞菌）。微阵列上的 31 种肽被注释为具有抗菌活性，其中 26 种在 pAF 中被检测到。胱抑素 C 和乳铁蛋白是 pAF 中表达最高的抗菌蛋白之一。ELISA 证实胱抑素 C 和乳铁蛋白与溶菌酶一起存在于 pAF 中。乳铁蛋白和溶菌酶的免疫沉淀减少了，但没有消除 pAF 的抗菌活性。
我们的数据表明，pAF 通过保留针对广泛的与伤口相关病原体的抗菌蛋白来保持抗菌活性。
