Frieser M, Hallmann R, Johansson S, Vestweber D, Goodman S L, Sorokin L
Institute for Experimental Medicine, University of Erlangen-Nürnberg, Germany.
Eur J Immunol. 1996 Dec;26(12):3127-36. doi: 10.1002/eji.1830261245.
The mechanism of adhesion of purified mouse polymorphonuclear granulocytes (PMN) to extracellular matrix proteins characteristic of basement membranes and the interstitium has been investigated and compared with the adhesion of a mouse progranulocytic cell line, 32DC13, and a mouse monocytic cell line, WEHI 78/24. All three cell types bound specifically to fibronectin and vitronectin to different degrees under different cellular activation states. 32DC13 bound to fibronectin and vitronectin strongly, and this binding increased upon cellular activation with phorbol 12-myristate-13-acetate (PMA) but not with formyl-Met-Leu-Phe. Only 32DC13 showed significant binding to laminin-1. By contrast, WEHI 78/24 and PMN bound only fibronectin and vitronectin; this binding was weak and was altered only marginally upon activation with PMA. In the case of WEHI 78/24, a slight increase in adhesion both to fibronectin and to vitronectin was observed after cellular activation with PMA, while PMN adhesion to both substrates was slightly reduced. The mechanism of binding to fibronectin and vitronectin was similar in the three cell types. The integrin alpha5 beta1 mediated fibronectin adhesion, demonstrating for the first time the existence of a functionally active beta1 integrin on mouse PMN. Vitronectin binding was mediated by alpha(v) beta3, as demonstrated by the ability of alpha(v)-specific cyclic L-Arg-L-Gly-L-Asp-D-Phe-L-Val (RGDfV) peptide (EMD66203), and anti-beta3 antibody to inhibit cell adhesion. 32DC13 adhesion to laminin-1 was via the alpha6 beta1 integrin. None of the three cell types tested bound to the basement membrane proteins collagen type IV and perlecan, or to the interstitial stromal constituents tenascin, collagen types I, V and VI. Interestingly, perlecan and collagen type IV were found to repel all three cell types. The relative inability of PMN, WEHI 78/24, and 32DC13 to bind to extracellular matrix proteins characteristic of basement membranes and their ability to bind inflammatory markers of the interstitium is discussed with respect to leukocyte extravasation processes.
已对纯化的小鼠多形核粒细胞（PMN）与基底膜和间质特有的细胞外基质蛋白的黏附机制进行了研究，并与小鼠前粒细胞系32DC13和小鼠单核细胞系WEHI 78/24的黏附情况进行了比较。在不同的细胞激活状态下，这三种细胞类型均以不同程度特异性结合纤连蛋白和玻连蛋白。32DC13与纤连蛋白和玻连蛋白紧密结合，在用佛波酯12-肉豆蔻酸酯-13-乙酸酯（PMA）而非甲酰甲硫氨酰亮氨酰苯丙氨酸激活细胞后，这种结合会增强。只有32DC13显示出与层粘连蛋白-1有显著结合。相比之下，WEHI 78/24和PMN仅结合纤连蛋白和玻连蛋白；这种结合较弱，在用PMA激活后仅略有改变。就WEHI 78/24而言，在用PMA激活细胞后，观察到其对纤连蛋白和玻连蛋白的黏附均略有增加，而PMN对这两种底物的黏附则略有降低。三种细胞类型与纤连蛋白和玻连蛋白的结合机制相似。整合素α5β1介导纤连蛋白黏附，首次证明小鼠PMN上存在功能活性β1整合素。如α（v）特异性环L-精氨酸-L-甘氨酸-L-天冬氨酸-D-苯丙氨酸-L-缬氨酸（RGDfV）肽（EMD66203）和抗β3抗体抑制细胞黏附的能力所证明，玻连蛋白结合由α（v）β3介导。32DC13对层粘连蛋白-1的黏附是通过α6β1整合素。所测试的三种细胞类型均未与基底膜蛋白IV型胶原和基底膜聚糖结合，也未与间质基质成分腱生蛋白、I型、V型和VI型胶原结合。有趣的是，发现基底膜聚糖和IV型胶原排斥所有三种细胞类型。关于白细胞外渗过程，讨论了PMN、WEHI 78/24和32DC13相对无法与基底膜特有的细胞外基质蛋白结合以及它们与间质炎症标志物结合的能力。
