Determination of hyaluronan molecular mass distribution in human breast milk.

Determination of hyaluronan molecular mass distribution in human breast milk.
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DOI:
10.1016/j.ab.2014.12.020
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发表时间:
2015-04-01
影响因子:
2.9
通讯作者:
Cowman, Mary K.
Cowman, Mary K.
中科院分区:
生物学4区
文献类型:
--
作者:
Yuan, Han;Amin, Ripal;Ye, Xin;de la Motte, Carol A.;Cowman, Mary K.

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人乳中的透明质酸(HA)通过TLR4-和cd44依赖的信号传导介导宿主对微生物感染的反应。HA的信令通常是特定于大小的。由于平均分子质量(M)为35 kDa的纯透明质酸可以引起肠上皮细胞的保护反应,因此有人提出人乳透明质酸可能具有低M成分的生物活性。在这里,我们报告了HA在20个独特捐赠者的人乳样本中的大小分布。建立了一种新的透明质酸分析方法,即利用离子交换(IEX)色谱法按粒径对透明质酸进行分离,并利用竞争酶联吸附试验(ELSA)对不同粒径的透明质酸进行定量。将牛奶HA分成4个组分时,M≤20 kDa、M≈20-60 kDa和M≈60-110 kDa的HA平均分别占总HA的1.5%、1.4%和2%。其余95%为HA, M≥110 kDa。对13个样品的高分子量HA的电泳分析显示几乎相同的M分布,平均M为~ 440 kDa。母乳中较高的HA成分被认为与CD44结合,并通过低HA成分增强人β -防御素2 (HBD2)的诱导。
Hyaluronan (HA) in human milk mediates host responses to microbial infection, via TLR4- and CD44-dependent signaling. Signaling by HA is generally size-specific. Because pure HA with average molecular mass (M) of 35 kDa can elicit a protective response in intestinal epithelial cells, it has been proposed that human milk HA may have a bioactive low M component. Here we report the size distribution of HA in human milk samples from twenty unique donors. A new method for HA analysis, employingion exchange (IEX) chromatography to fractionate HA by size, and specific quantification of each size fraction by competitive Enzyme Linked Sorbent Assay (ELSA), was developed. When separated into four fractions, milk HA with M ≤ 20 kDa, M ≈20-60 kDa, and M ≈ 60-110 kDa comprised an average of 1.5%, 1.4% and 2% of the total HA, respectively. The remaining 95% was HA with M≥110 kDa. Electrophoretic analysis of the higher M HA from thirteen samples showed nearly identical M distributions, with an average M of ∼440 kDa. This higher M HA component in human milk is proposed to bind to CD44 and to enhance human beta defensin 2 (HBD2) induction by the low M HA components.
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