Meng H, Marchese M J, Garlick J A, Jelaska A, Korn J H, Gailit J, Clark R A, Gruber B L
Department of Medicine, School of Medicine, SUNY at Stony Brook 11794-8161, USA.
J Invest Dermatol. 1995 Dec;105(6):789-96. doi: 10.1111/1523-1747.ep12326075.
The capacity of mast cell products to mediate T-cell adhesion to fibroblasts was explored using heterotypic coculture systems or by exposing fibroblasts to mast-cell-conditioned media (MCCM), prepared by degranulating mast cells with calcium ionophore. Experimental results indicated that fibroblasts exposed to MCCM for 24 h bound fivefold more T cells than control fibroblasts. Binding was inhibited with intercellular adhesion molecule-1 (ICAM-1) or vascular cell adhesion molecule-1 (VCAM-1) neutralizing antibodies. Enzyme-linked immunosorbent assay and fluorescence-activated cell sorter analysis revealed that fibroblasts exposed to MCCM markedly increased ICAM-1 and VCAM-1 surface expression by 4 h, with levels maximal at 16 h and returning toward baseline by 48 h. A dose-dependent response of ICAM-1 and VCAM-1 expression was noted using serial dilutions of MCCM or by altering the ratio of degranulated mast cells cocultured with fibroblasts. Similar results were obtained using human fibroblasts derived from the dermis, synovium, and lung, although lung fibroblasts were generally less responsive. Northern analysis confirmed that MCCM regulated ICAM-1 and VCAM-1 expression at the mRNA level. In summary, mast cell products stimulated fibroblast surface expression, steady-state mRNA levels, and functional expression of ICAM-1 and VCAM-1. Experimental data suggest that mast-cell-derived tumor necrosis factor-alpha may be in large part responsible for these observations, although further studies using human mast cells will be required. Using a skin-equivalent organotypic coculture model with fibroblasts admixed with mast cells, we observed increased ICAM-1 expression in both keratinocytes and fibroblasts after activation of the mast cells.
利用异型共培养系统或通过将成纤维细胞暴露于用钙离子载体使肥大细胞脱颗粒制备的肥大细胞条件培养基(MCCM),探讨了肥大细胞产物介导T细胞与成纤维细胞黏附的能力。实验结果表明,暴露于MCCM 24小时的成纤维细胞结合的T细胞数量是对照成纤维细胞的五倍。细胞间黏附分子-1(ICAM-1)或血管细胞黏附分子-1(VCAM-1)中和抗体可抑制这种结合。酶联免疫吸附测定和荧光激活细胞分选分析显示,暴露于MCCM的成纤维细胞在4小时时ICAM-1和VCAM-1表面表达明显增加,16小时时达到最高水平,48小时时恢复至基线水平。使用MCCM系列稀释液或改变与成纤维细胞共培养的脱颗粒肥大细胞比例时,观察到ICAM-1和VCAM-1表达呈剂量依赖性反应。使用源自真皮、滑膜和肺的人成纤维细胞也获得了类似结果,尽管肺成纤维细胞的反应通常较弱。Northern分析证实,MCCM在mRNA水平上调节ICAM-1和VCAM-1的表达。总之,肥大细胞产物刺激了成纤维细胞表面表达、稳态mRNA水平以及ICAM-1和VCAM-1的功能性表达。实验数据表明,肥大细胞衍生的肿瘤坏死因子-α可能在很大程度上导致了这些观察结果,尽管还需要使用人肥大细胞进行进一步研究。使用成纤维细胞与肥大细胞混合的皮肤等效器官型共培养模型,我们观察到肥大细胞激活后角质形成细胞和成纤维细胞中ICAM-1表达均增加。