Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Stockholm, Sweden.
Cell Mol Life Sci. 2021 Jun;78(12):5197-5212. doi: 10.1007/s00018-021-03858-y. Epub 2021 May 22.
Multiple cellular processes, such as immune responses and cancer cell metastasis, crucially depend on interconvertible migration modes. However, knowledge is scarce on how infectious agents impact the processes of cell adhesion and migration at restrictive biological barriers. In extracellular matrix, dendritic cells (DCs) infected by the obligate intracellular protozoan Toxoplasma gondii undergo mesenchymal-to-amoeboid transition (MAT) for rapid integrin-independent migration. Here, in a cellular model of the blood-brain barrier, we report that parasitised DCs adhere to polarised endothelium and shift to integrin-dependent motility, accompanied by elevated transendothelial migration (TEM). Upon contact with endothelium, parasitised DCs dramatically reduced velocities and adhered under both static and shear stress conditions, thereby obliterating the infection-induced amoeboid motility displayed in collagen matrix. The motility of adherent parasitised DCs on endothelial monolayers was restored by blockade of β1 and β2 integrins or ICAM-1, which conversely reduced motility on collagen-coated surfaces. Moreover, parasitised DCs exhibited enhanced translocation across highly polarised primary murine brain endothelial cell monolayers. Blockade of β1, β2 integrins, ICAM-1 and PECAM-1 reduced TEM frequencies. Finally, gene silencing of the pan-integrin-cytoskeleton linker talin (Tln1) or of β1 integrin (Itgb1) in primary DCs resulted in increased motility on endothelium and decreased TEM. Adding to the paradigms of leukocyte diapedesis, the findings provide novel insights in how an intracellular pathogen impacts the migratory plasticity of leukocytes in response to the cellular environment, to promote infection-related dissemination.
多种细胞过程,如免疫反应和癌细胞转移,严重依赖于可相互转换的迁移模式。然而,对于传染性病原体如何影响细胞黏附和迁移在限制生物屏障中的过程,我们的了解甚少。在细胞外基质中,被专性细胞内原生动物刚地弓形虫感染的树突状细胞(DC)会发生间质到阿米巴样的转变(MAT),以实现快速的整联蛋白非依赖性迁移。在这里,在血脑屏障的细胞模型中,我们报告感染的 DC 与极化的内皮细胞黏附,并转变为整联蛋白依赖性运动,同时伴随跨内皮迁移(TEM)的增加。与内皮接触后,感染的 DC 显著降低速度,并在静态和切应力条件下黏附,从而消除了在胶原蛋白基质中观察到的感染诱导的阿米巴样运动。在内皮单层上,通过阻断β1 和β2 整合素或 ICAM-1 可以恢复黏附感染的 DC 的运动,而这反过来又降低了在胶原蛋白涂层表面的运动。此外,感染的 DC 表现出增强穿过高度极化的原代小鼠脑内皮细胞单层的迁移能力。阻断β1、β2 整合素、ICAM-1 和 PECAM-1 减少了 TEM 频率。最后,用小干扰 RNA 沉默泛整合素-细胞骨架连接蛋白 talin(Tln1)或β1 整合素(Itgb1)在原代 DC 中,导致在内皮上的迁移能力增加,而 TEM 减少。除了白细胞穿出的范例外,这些发现提供了新的见解,即细胞内病原体如何影响白细胞的迁移可塑性,以响应细胞环境,促进与感染相关的传播。