Nakayama F, Nishihara S, Iwasaki H, Kudo T, Okubo R, Kaneko M, Nakamura M, Karube M, Sasaki K, Narimatsu H
Division of Cell Biology, Institute of Life Science, Soka University, Tokyo 192-8577, Japan.
J Biol Chem. 2001 May 11;276(19):16100-6. doi: 10.1074/jbc.M007272200. Epub 2001 Feb 23.
The CD15 carbohydrate epitope is expressed in mature human neutrophils, monocytes, and promyelocytes. We aimed to determine the alpha1,3-fucosyltransferase responsible for the expression of CD15 in each subpopulation of leukocytes. Three alpha1,3-fucosyltransferases, FUT4, FUT7, and FUT9, are expressed in human leukocytes. We demonstrated that FUT9 exhibits 20-fold stronger activity for CD15 synthesis than FUT4, whereas FUT4 exhibits 4.5-fold stronger activity for CDw65 synthesis than FUT9. By competitive reverse transcriptase-polymerase chain reaction, FUT9 was found to be strongly expressed in mature granulocytes and peripheral blood mononuclear cell, but not in monocytes. CD34(+) and CD15(+) cells in cord blood and myeloid cell lines (HL-60 and U937) did not express FUT9 at all. FUT4 transcripts were ubiquitously expressed in all blood cells and all cultured cell lines, with HL-60 and U937 cells in particular expressing a number of FUT4 transcripts. Transfection of the FUT9 gene into Jurkat and U937 cells demonstrated that FUT9 has the potential to express CD15 in myeloid and lymphoid cells. These findings suggest that the expression of CD15 in mature granulocytes is directed by FUT9, whereas it is determined in promyelocytes and monocytes by FUT4. Measurement of CD15 synthesizing activity in cell homogenates of each cell population using the polylactosamine acceptor further supported these conclusions.
CD15碳水化合物表位在成熟的人类中性粒细胞、单核细胞和早幼粒细胞中表达。我们旨在确定负责白细胞各亚群中CD15表达的α1,3-岩藻糖基转移酶。三种α1,3-岩藻糖基转移酶,即FUT4、FUT7和FUT9,在人类白细胞中表达。我们证明,FUT9对CD15合成的活性比FUT4强20倍,而FUT4对CDw65合成的活性比FUT9强4.5倍。通过竞争性逆转录聚合酶链反应发现,FUT9在成熟粒细胞和外周血单个核细胞中强烈表达,但在单核细胞中不表达。脐血中的CD34(+)和CD15(+)细胞以及髓系细胞系(HL-60和U937)根本不表达FUT9。FUT4转录本在所有血细胞和所有培养细胞系中普遍表达,尤其是HL-60和U937细胞表达大量FUT4转录本。将FUT9基因转染到Jurkat和U937细胞中表明,FUT9有潜力在髓系和淋巴细胞中表达CD15。这些发现表明,成熟粒细胞中CD15的表达由FUT9指导,而在早幼粒细胞和单核细胞中则由FUT4决定。使用聚乳糖胺受体测量每个细胞群体的细胞匀浆中CD15合成活性进一步支持了这些结论。