Screening and identification of hepatitis B surface antigen binding to lactoferrin in human milk

Q4 Medicine

中华围产医学杂志 Pub Date : 2019-07-16 DOI:10.3760/CMA.J.ISSN.1007-9408.2019.07.006

Zhao-hua Zhang, Jingli Liu, Jing Feng, Y. Dai, Yali Hu, Yihua Zhou

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引用次数: 0

Abstract

Objective Human milk of mothers with positive hepatitis B surface antigen (HBsAg) contains hepatitis B virus (HBV). However, breastfeeding does not increase the risk of mother-to-infant transmission of HBV. Previous investigations demonstrated that breast milk has a property of binding with HBsAg. This study aimed to identify the component in human milk that can bind to HBsAg. Methods This study was performed in Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School, from June 2015 to February 2017. Human milk samples from two postpartum women with negative HBV markers and two control samples of cow milk and goat milk were analyzed by Far-Western blot, in which highly purified recombinant yeast HBsAg was used to bind with whey proteins. Based on the results of mass-spectrum analysis, competition inhibition test was used to confirm the functioning component. Results Far-Western blot showed remarkable protein bands at the relative molecular weight of about 80 000 in both lanes of human milk, but none in the lane of cow or goat milk. Mass-spectrum analysis of the protein band indicated there were proteins sharing 28.4%-93.4% homology in amino acid sequences with five proteins with the highest homology to lactoferrin (93.4%). Further Far-Western blot with purified recombinant lactoferrin showed that lactoferrin could bind to the recombinant HBsAg. Competition inhibition test suggested that the purified recombinant lactoferrin inhibited the binding of HBsAg to its antibody in a dose-dependent manner. Conclusions This study confirms the capability of lactoferrin in human milk to combine with HBsAg, suggesting that lactoferrin can bind to HBV. Further study on whether lactoferrin can inhibit the infectivity of HBV would be valuable to clarify the reason for not increasing the risk of mother-to-infant transmission of HBV by breastfeeding. Key words: Milk, human; Lactoferrin; Hepatitis B surface antigens

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母乳中与乳铁蛋白结合的乙型肝炎表面抗原的筛选与鉴定

目的乙型肝炎表面抗原（HBsAg）阳性母亲母乳中含有乙型肝炎病毒（HBV）。然而，母乳喂养不会增加母婴传播HBV的风险。先前的研究表明，母乳具有与HBsAg结合的特性。本研究旨在鉴定母乳中能与HBsAg结合的成分。方法本研究于2015年6月至2017年2月在南京大学医学院附属医院南京鼓楼医院进行。用高纯度重组酵母HBsAg与乳清蛋白结合的Far Western印迹分析了两名HBV标志物阴性的产后妇女的母乳样本和两份对照牛奶和山羊奶样本。基于质谱分析的结果，采用竞争抑制试验来确认功能成分。结果Far Western印迹在母乳和羊奶中均显示出相对分子量约为8万的显著蛋白质条带，而在牛奶和山羊奶中则没有。蛋白质条带的质谱分析表明，与乳铁蛋白同源性最高的5种蛋白质（93.4%）的氨基酸序列同源性为28.4%～93.4%，进一步用纯化的重组乳铁蛋白进行远蛋白印迹，乳铁蛋白能与重组HBsAg结合。竞争抑制试验表明，纯化的重组乳铁蛋白以剂量依赖的方式抑制HBsAg与其抗体的结合。结论本研究证实了母乳中乳铁蛋白与HBsAg结合的能力，提示乳铁蛋白可以与HBV结合。进一步研究乳铁蛋白是否能抑制乙型肝炎病毒的传染性，有助于阐明母乳喂养不会增加母婴传播乙型肝炎病毒风险的原因。关键词：牛奶，人类；乳铁蛋白；乙型肝炎表面抗原

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来源期刊

中华围产医学杂志 Medicine-Obstetrics and Gynecology

CiteScore

0.70

自引率

0.00%

发文量

4446

期刊介绍： Chinese Journal of Perinatal Medicine was founded in May 1998. It is one of the journals of the Chinese Medical Association, which is supervised by the China Association for Science and Technology, sponsored by the Chinese Medical Association, and hosted by Peking University First Hospital. Perinatal medicine is a new discipline jointly studied by obstetrics and neonatology. The purpose of this journal is to "prenatal and postnatal care, improve the quality of the newborn population, and ensure the safety and health of mothers and infants". It reflects the new theories, new technologies, and new progress in perinatal medicine in related disciplines such as basic, clinical and preventive medicine, genetics, and sociology. It aims to provide a window and platform for academic exchanges, information transmission, and understanding of the development trends of domestic and foreign perinatal medicine for the majority of perinatal medicine workers in my country.