Podosomes, But Not the Maturation Status, Determine the Protease-Dependent 3D Migration in Human Dendritic Cells.
Podosomes, But Not the Maturation Status, Determine the Protease-Dependent 3D Migration in Human Dendritic Cells.
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DOI：
10.3389/fimmu.2018.00846
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发表时间：
2018
影响因子：
7.3
通讯作者：
Maridonneau-Parini I
中科院分区：
文献类型：
--
作者：
Cougoule C;Lastrucci C;Guiet R;Mascarau R;Meunier E;Lugo-Villarino G;Neyrolles O;Poincloux R;Maridonneau-Parini I
树突状细胞（DC）是专业的抗原呈递细胞，分散在抗原暴露的组织和引流淋巴结中，并调查身体的病原体。它们在组织（3D环境）中迁移的能力对于有效的免疫反应至关重要。感染后，Toll样受体（TLR）识别病原体相关分子模式（PAMP）触发DC成熟。成熟的DC（mDC）在体内或体外在胶原基质中基本上使用蛋白酶非依赖性、ROCK依赖性的变形样模式。然而，人类未成熟DC（iDC）使用的3D迁移机制仍然很差。在这里，我们揭示了人类单核细胞衍生的DC能够在3D中使用两种迁移模式。在纤维胶原I的多孔基质中，iDC采用变形虫迁移模式。在凝胶化胶原I或Matrigel的致密基质中，iDC使用蛋白酶依赖性、ROCK非依赖性间充质迁移模式。在LPS激活TLR 4后，mDC-LPS失去形成足体和降解基质的能力，沿着间充质迁移受损。通过Pam 3CSK 4激活TLR 2导致DC成熟、足体维持和有效的间充质迁移。在所有这些条件下，当DC在致密基质中使用间充质模式时，它们在细胞突起的尖端形成3D podosomes。使用PGE 2，已知破坏树突状细胞中的podosomes，我们观察到细胞保持在未成熟状态，间充质迁移模式被取消。我们还观察到，虽然CCL 5（iDC的引诱剂）增强iDC的变形样和间充质迁移，但CCL 19和CCL 21（mDC的引诱剂）仅增强mDC-LPS变形样迁移而不触发间充质迁移。最后，我们研究了iDC在肿瘤细胞球体（一种组织样3D环境）中的迁移。我们观察到iDC使用两种迁移模式浸润肿瘤细胞的球体。总之，这些结果表明，人DC采用间充质模式在3D致密环境中迁移，这依赖于它们形成独立于其成熟状态的podosomes的能力，为进一步研究DC在致密组织中的体内迁移及其在感染期间的调节铺平了道路。
Dendritic cells (DC) are professional Antigen-Presenting Cells scattered throughout antigen-exposed tissues and draining lymph nodes, and survey the body for pathogens. Their ability to migrate through tissues, a 3D environment, is essential for an effective immune response. Upon infection, recognition of Pathogen-Associated Molecular Patterns (PAMP) by Toll-like receptors (TLR) triggers DC maturation. Mature DC (mDC) essentially use the protease-independent, ROCK-dependent amoeboid mode in vivo, or in collagen matrices in vitro. However, the mechanisms of 3D migration used by human immature DC (iDC) are still poorly characterized. Here, we reveal that human monocyte-derived DC are able to use two migration modes in 3D. In porous matrices of fibrillar collagen I, iDC adopted the amoeboid migration mode. In dense matrices of gelled collagen I or Matrigel, iDC used the protease-dependent, ROCK-independent mesenchymal migration mode. Upon TLR4 activation by LPS, mDC-LPS lose the capacity to form podosomes and degrade the matrix along with impaired mesenchymal migration. TLR2 activation by Pam3CSK4 resulted in DC maturation, podosome maintenance, and efficient mesenchymal migration. Under all these conditions, when DC used the mesenchymal mode in dense matrices, they formed 3D podosomes at the tip of cell protrusions. Using PGE2, known to disrupt podosomes in DC, we observed that the cells remained in an immature status and the mesenchymal migration mode was abolished. We also observed that, while CCL5 (attractant of iDC) enhanced both amoeboid and mesenchymal migration of iDC, CCL19 and CCL21 (attractants of mDC) only enhanced mDC-LPS amoeboid migration without triggering mesenchymal migration. Finally, we examined the migration of iDC in tumor cell spheroids, a tissue-like 3D environment. We observed that iDC infiltrated spheroids of tumor cells using both migration modes. Altogether, these results demonstrate that human DC adopt the mesenchymal mode to migrate in 3D dense environments, which relies on their capacity to form podosomes independent of their maturation status, paving the way of further investigations on in vivo DC migration in dense tissues and its regulation during infections.