Delay of migrating leukocytes by the basement membrane deposited by endothelial cells in long-term culture
Delay of migrating leukocytes by the basement membrane deposited by endothelial cells in long-term culture
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DOI：
10.1016/j.yexcr.2010.10.022
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发表时间：
2011-02-01
影响因子：
3.7
通讯作者：
Nash, Gerard B.
中科院分区：
文献类型：
--
作者：
Burton, Victoria J.;Butler, Lynn M.;Nash, Gerard B.
我们研究了人类白细胞通过内皮细胞（EC），特别是其底层的基底膜（BM）的迁移。EC在3 μ m孔过滤器或胶原凝胶上培养20天以形成不同的BM，然后用肿瘤坏死因子-α、白细胞介素-1 β或干扰素-γ处理。与4天培养物相比，神经元迁移通过甘氨酸处理的EC和BM延迟20天。单独的BM阻碍中性粒细胞的趋化性，如果新鲜的EC被简单地培养在剥离的BM上，则在迁移中再次出现停滞。在淋巴细胞和单核细胞的研究中，我们可以检测到在基于过滤器或凝胶的模型中，20天培养物与4天培养物的迁移延迟很小。直接显微镜观察表明，BM也举行了流动条件下的中性粒细胞迁移。用抗整联蛋白抗体处理滤器的上室和/或下室，表明中性粒细胞通过内皮单层的迁移依赖于β 2-整联蛋白，而不是β 1-或β 3-整联蛋白。对于所有培养物，β（1）-和β（2）-整联蛋白支持从内皮下室的迁移，但阻断β（3）-整联蛋白仅有效抑制20天培养物的迁移。流式细胞术表明，在中性粒细胞迁移过程中，β（1）-或β（3）-整合素的表达没有净增加，它们的特异性内皮下功能可能依赖于迁移过程中整合素的周转。这些研究表明，BM是人类嗜中性粒细胞迁移的独特屏障，并且β（3）-整联蛋白在穿过该屏障中特别重要。BM对淋巴细胞和单核细胞的影响较小，这支持了这样一个概念，即穿过BM是一个单独的、白细胞特异性的、受调节的迁移步骤。(C)2010年爱思唯尔公司All rights reserved.
We investigated the migration of human leukocytes through endothelial cells (EC), and particularly their underlying basement membrane (BM). EC were cultured for 20 days on 3 mu m-pore filters or collagen gels to form a distinct BM, and then treated with tumour necrosis factor-alpha, interleukin-1 beta or interferon-gamma. Neutrophil migration through the cytokine-treated EC and BM was delayed for 20-day compared to 4-day cultures. The BM alone obstructed chemotaxis of neutrophils, and if fresh EC were briefly cultured on stripped BM, there was again a hold-up in migration. In studies with lymphocytes and monocytes, we could detect little hold-up of migration for 20-day versus 4-day cultures, in either the filter- or gel-based models. Direct microscopic observations showed that BM also held-up neutrophil migration under conditions of flow. Treatment of upper and/or lower compartments of filters with antibodies against integrins, showed that neutrophil migration through the endothelial monolayer was dependent on beta(2)-integrins, but not beta 1- or beta(3)-integrins. Migration from the subendothelial compartment was supported by beta(1)- and beta(2)-integrins for all cultures, but blockade of beta(3)-integrin only inhibited migration effectively for 20-day cultures. Flow cytometry indicated that there was no net increase in expression of beta(1)- or beta(3)-integrins during neutrophil migration, and that their specific subendothelial function was likely dependent on turnover of integrins during migration. These studies show that BM is a distinct barrier to migration of human neutrophils, and that beta(3)-integrins are particularly important in crossing this barrier. The lesser effect of BM on lymphocytes and monocytes supports the concept that crossing the BM is a separate, leukocyte-specific, regulated step in migration. (C) 2010 Elsevier Inc. All rights reserved.