Higashi Nobuaki, Morikawa Akiko, Fujioka Kouki, Fujita Yuko, Sano Yoshihiko, Miyata-Takeuchi Megumi, Suzuki Noriko, Irimura Tatsuro
Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Int Immunol. 2002 Jun;14(6):545-54. doi: 10.1093/intimm/dxf021.
We studied the expression of a human macrophage lectin specific for galactose/N-acetylgalactosamine (hMGL) during macrophage differentiation. The expression of hMGL during the in vitro differentiation induced by human serum was examined by immunostaining and Western blotting with a specific mAb, MLD-1, as well as with RT-PCR analysis. hMGL was detected on cells at an intermediate stage of differentiation. These cells were round, slightly larger in size (12.7 +/- 0.2 microm) than monocytes (9.8 +/- 0.1 microm) and expressed the macrophage marker CD14, but lacked the dendritic cell marker CD1a. The highest levels of expression occurred after 2-4 days of culture. At this time point, MLD-1 prominently stained 20-40% of the cells. Monocytes cultured for 16 h or fully differentiated monocyte-derived macrophages were negative or weak for hMGL expression. Similar transient expression was also observed during granulocyte macrophage colony stimulating factor- or macrophage colony stimulating factor-dependent macrophage differentiation. The lectin was characterized as a functional endocytic receptor for glycosylated macromolecules, since the uptake of carbohydrate polymers was partially inhibited by the addition of MLD-1. The distribution of hMGL(+) cells in normal human skin was found by immunostaining to be mainly in the upper dermis distant from vascular structures. More than 90% of the hMGL(+) cells were double stained with anti-CD68 mAb and constituted approximately 20% of the CD68(+) cells. We suggest that the dermal hMGL(+) cells are a subset of differentiated cells derived from monocytes and that hMGL is a unique marker for cells at an intermediate stage of macrophage differentiation.
我们研究了半乳糖/N-乙酰半乳糖胺特异性人巨噬细胞凝集素（hMGL）在巨噬细胞分化过程中的表达。通过免疫染色、使用特异性单克隆抗体MLD-1进行蛋白质印迹以及逆转录聚合酶链反应（RT-PCR）分析，检测了人血清诱导的体外分化过程中hMGL的表达。在分化的中间阶段细胞上检测到了hMGL。这些细胞呈圆形，大小（12.7±0.2微米）略大于单核细胞（9.8±0.1微米），表达巨噬细胞标志物CD14，但缺乏树突状细胞标志物CD1a。培养2 - 4天后表达水平最高。此时，MLD-1可显著染色20% - 40%的细胞。培养16小时的单核细胞或完全分化的单核细胞衍生巨噬细胞hMGL表达为阴性或弱阳性。在粒细胞巨噬细胞集落刺激因子或巨噬细胞集落刺激因子依赖的巨噬细胞分化过程中也观察到了类似的瞬时表达。该凝集素被鉴定为糖基化大分子的功能性内吞受体，因为添加MLD-1可部分抑制碳水化合物聚合物的摄取。通过免疫染色发现，正常人类皮肤中hMGL(+)细胞主要分布在远离血管结构的真皮上层。超过90%的hMGL(+)细胞与抗CD68单克隆抗体双染，约占CD68(+)细胞的20%。我们认为真皮hMGL(+)细胞是源自单核细胞的分化细胞亚群，并且hMGL是巨噬细胞分化中间阶段细胞的独特标志物。
