Microarray analysis of human milk cells: persistent high expression of osteopontin during the lactation period
Microarray analysis of human milk cells: persistent high expression of osteopontin during the lactation period
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DOI：
10.1111/j.1365-2249.2004.02549.x
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发表时间：
2004-10-01
影响因子：
4.6
通讯作者：
Hara, T
中科院分区：
文献类型：
--
作者：
Nagatomo, T;Ohga, S;Hara, T
为了继续探索母乳的免疫学作用，对人乳细胞进行了细胞因子和生长因子的基因芯片分析。在240个细胞因子相关基因中，骨桥蛋白(OPN)基因的表达最高。实时定量聚合酶链式反应显示，初乳细胞中OPN基因的表达水平比PHA刺激的外周血单个核细胞(PBMNCs)高约100倍，比外周血CD14(+)细胞高10000倍。早乳或成熟乳细胞中OPN mRNA的中位数是初乳细胞的3倍以上。Western印迹分析表明，人乳中有OPN的全长和短型蛋白表达。用酶联免疫吸附试验检测，早乳或成熟乳清中的全长OPN浓度继续高于初乳、乳清和血浆中的浓度。早期乳汁(产后3~7天)的OPN蛋白含量最高，而晚期成熟乳细胞(产后1年)的OPN mRNA表达水平最高。免疫组织化学和免疫细胞化学染色结果表明，在分泌活跃的乳腺组织中可见产生骨桥蛋白的上皮细胞和巨噬细胞。这些结果表明，OPN在母乳细胞中的持续异常高表达在母乳喂养婴儿的免疫发育中起着潜在的作用。
To continue the search for immunological roles of breast milk, cDNA microarray analysis on cytokines and growth factors was performed for human milk cells. Among the 240 cytokine-related genes, osteopontin (OPN) gene ranked top of the expression. Real-time PCR revealed that the OPN mRNA levels in colostrum cells were approximately 100 times higher than those in PHA-stimulated peripheral blood mononuclear cells (PBMNCs), and 10 000 times higher than those in PB CD14(+) cells. The median levels of OPN mRNA in early milk or mature milk cells were more than three times higher than those in colostrum cells. Western blot analysis of human milk showed appreciable expression of full-length and short form proteins of OPN. The concentrations of full-length OPN in early milk or mature milk whey continued to be higher than those in colostrum whey and plasma as assessed by ELISA. The early milk (3-7 days postpartum) contained the highest concentrations of OPN protein, while the late mature milk cells (1 years postpartum) had the highest expression of OPN mRNA of all the lactating periods. The results of immunohistochemical and immunocytochemical staining indicated that OPN-producing epithelial cells and macrophages are found in actively lactating mammary glands. These results suggest that the persistently and extraordinarily high expression of OPN in human milk cells plays a potential role in the immunological development of breast-fed infants.