Baricelli Joanna, Rocafull Miguel A, Vázquez Desiree, Bastidas Betsi, Báez-Ramirez Estalina, Thomas Luz E
Laboratorio de Fisiología Molecular, Centro de Biofísica y Bioquímica, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, Venezuela; Banco de Leche Humana, Hospital Universitario de Caracas, Caracas, Venezuela.
Banco de Leche Humana, Hospital Universitario de Caracas, Caracas, Venezuela; Laboratorio Delgado Launois, Clínica Lugo, Maracay, Venezuela.
J Pediatr (Rio J). 2015 Jan-Feb;91(1):36-43. doi: 10.1016/j.jped.2014.05.006. Epub 2014 Sep 7.
To describe the antimicrobial activity of β-defensin-2 produced in the mammary gland and secreted in human breast milk.
The peptide production was performed by DNA cloning. β-defensin-2 levels were quantified in 61 colostrum samples and 39 mature milk samples from healthy donors, by an indirect enzyme-linked immunosorbent assay (ELISA). Using halo inhibition assay, this study assessed activity against seven clinical isolates from diarrheal feces of children between 0 and 2 years of age. The activity of β-defensin-2 against three opportunistic pathogens that can cause nosocomial infections was determined by microdilution test.
The peptide levels were higher in colostrum (n=61) than in mature milk samples (n=39), as follows: median and range, 8.52 (2.6-16.3) μg/ml versus 0.97 (0.22-3.78), p<0.0001; Mann-Whitney test. The recombinant peptide obtained showed high antimicrobial activity against a broad range of pathogenic bacteria. Its antibacterial activity was demonstrated in a disk containing between 1-4 μg, which produced inhibition zones ranging from 18 to 30 mm against three isolates of Salmonella spp. and four of E. coli. β-defensin-2 showed minimum inhibitory concentrations (MICs) of 0.25 μg/mL and 0.5 μg/mL for S. marcescen and P. aeruginosa, respectively, while a higher MIC (4 μg/mL) was obtained against an isolated of multidrug-resistant strain of A. baumannii.
To the authors' knowledge, this study is the first to report β-defensin-2 levels in Latin American women. The production and the activity of β-defensin-2 in breast milk prove its importance as a defense molecule for intestinal health in pediatric patients.
描述乳腺产生并分泌于人母乳中的β-防御素-2的抗菌活性。
通过DNA克隆进行肽的生产。采用间接酶联免疫吸附测定法(ELISA)对61份来自健康供体的初乳样本和39份成熟乳样本中的β-防御素-2水平进行定量。本研究使用光晕抑制试验评估了对7株来自0至2岁儿童腹泻粪便的临床分离株的活性。通过微量稀释试验测定了β-防御素-2对三种可引起医院感染的机会性病原体的活性。
初乳(n = 61)中的肽水平高于成熟乳样本(n = 39),如下所示:中位数和范围,8.52(2.6 - 16.3)μg/ml对0.97(0.22 -
3.78)μg/ml,p < 0.0001;曼-惠特尼检验。获得的重组肽对多种病原菌显示出高抗菌活性。其抗菌活性在含有1 - 4μg的圆盘中得到证明,该圆盘对三种沙门氏菌属分离株和四种大肠杆菌分离株产生的抑菌圈范围为18至30mm。β-防御素-2对粘质沙雷氏菌和铜绿假单胞菌的最低抑菌浓度(MIC)分别为0.25μg/mL和0.5μg/mL,而对一株鲍曼不动杆菌多重耐药菌株的MIC较高(4μg/mL)。
据作者所知,本研究是首次报告拉丁美洲女性中β-防御素-2水平。母乳中β-防御素-2的产生和活性证明了其作为儿科患者肠道健康防御分子的重要性。