Singer I I, Scott S, Kawka D W, Kazazis D M
Department of Biochemical and Molecular Pathology, Merck, Sharp & Dohme Research Laboratories, Merck & Co., Inc., Rahway, New Jersey 07065.
J Cell Biol. 1989 Dec;109(6 Pt 1):3169-82. doi: 10.1083/jcb.109.6.3169.
We have localized several major extracellular matrix protein receptors in the specific granules of human polymorphonuclear (PMN) and monocytic leukocytes using double label immunoelectron microscopy (IEM) with ultrathin frozen sections and colloidal-gold conjugates. Rabbit antibodies to 67-kD human laminin receptor (LNR) were located on the inner surface of the specific granule membrane and within its internal matrix. LNR antigens co-distributed with lactoferrin, a marker of specific granules, but did not co-localize with elastase in azurophilic granules of PMNs. Further, CD11b/CD18 (leukocyte receptor for C3bi, fibrinogen, endothelial cells, and endotoxin), mammalian fibronectin receptor (FNR), and vitronectin receptor (VNR) antigens were also co-localized with LNR in PMN specific granules. A similar type of granule was found in monocytes which stained for LNR, FNR, VNR, CD18, and lysozyme. Activation of PMNs with either PMA, f-met-leu-phe (fMLP), tumor necrosis factor (TNF), or monocytic leukocytes with lipopolysaccharide (LPS), induced fusion of specific granules with the cell membrane and expression of both LNR and CD18 antigens on the outer cell surface. Further, stimulation led to augmented PMN adhesion on LN substrata, and six- to eightfold increases in specific binding of soluble LN that was inhibited by LNR antibody. These results indicate that four types of extracellular matrix receptors are located in leukocyte specific granules, and suggest that up-regulation of these receptors during inflammation may mediate leukocyte adhesion and extravasation. We have thus termed leukocyte specific granules adhesomes.
我们使用超薄冰冻切片和胶体金缀合物的双标记免疫电子显微镜(IEM),在人多形核(PMN)和单核白细胞的特定颗粒中定位了几种主要的细胞外基质蛋白受体。针对67-kD人层粘连蛋白受体(LNR)的兔抗体位于特定颗粒膜的内表面及其内部基质中。LNR抗原与乳铁蛋白(一种特定颗粒的标志物)共分布,但在PMN的嗜天青颗粒中不与弹性蛋白酶共定位。此外,CD11b/CD18(C3bi、纤维蛋白原、内皮细胞和内毒素的白细胞受体)、哺乳动物纤连蛋白受体(FNR)和玻连蛋白受体(VNR)抗原也在PMN特定颗粒中与LNR共定位。在单核细胞中发现了一种类似类型的颗粒,其对LNR、FNR、VNR、CD18和溶菌酶染色。用佛波酯(PMA)、N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸(f-met-leu-phe,fMLP)、肿瘤坏死因子(TNF)激活PMN,或用脂多糖(LPS)激活单核白细胞,可诱导特定颗粒与细胞膜融合,并在细胞外表面表达LNR和CD18抗原。此外,刺激导致PMN在层粘连蛋白(LN)基质上的黏附增强,可溶性LN的特异性结合增加6至8倍,这被LNR抗体抑制。这些结果表明,四种类型的细胞外基质受体位于白细胞特定颗粒中,并提示炎症期间这些受体的上调可能介导白细胞黏附和外渗。因此,我们将白细胞特定颗粒称为黏附体。