Department of Immunology, The Weizmann Institute of Science, Rehovot, Israel.
Faculty of Engineering, Bar Ilan Institute of Nanotechnologies and Advanced Materials, Bar Ilan University, Ramat Gan, Israel.
J Leukoc Biol. 2018 Aug;104(2):239-251. doi: 10.1002/JLB.3HI1217-488R. Epub 2018 Mar 30.
Transendothelial migration (TEM) of lymphocytes and neutrophils is associated with the ability of their deformable nuclei to displace endothelial cytoskeletal barriers. Lamin A is a key intermediate filament component of the nuclear lamina that is downregulated during granulopoiesis. When elevated, lamin A restricts nuclear squeezing through rigid confinements. To determine if the low lamin A expression by leukocyte nuclei is critical for their exceptional squeezing ability through endothelial barriers, we overexpressed this protein in granulocyte-like differentiated HL-60 cells. A 10-fold higher lamin A expression did not interfere with chemokinetic motility of these granulocytes on immobilized CXCL1. Furthermore, these lamin A high leukocytes exhibited normal chemotaxis toward CXCL1 determined in large pore transwell barriers, but poorly squeezed through 3 μm pores toward identical CXCL1 gradients. Strikingly, however, these leukocytes successfully completed paracellular TEM across inflamed endothelial monolayers under shear flow, albeit with a small delay in nuclear squeezing into their sub-endothelial pseudopodia. In contrast, CXCR2 mediated granulocyte motility through collagen I barriers was dramatically delayed by lamin A overexpression due to a failure of lamin A high nuclei to translocate into the pseudopodia of the granulocytes. Collectively, our data predict that leukocytes maintain a low lamin A content in their nuclear lamina in order to optimize squeezing through extracellular collagen barriers but can tolerate high lamin A content when crossing the highly adaptable barriers presented by the endothelial cytoskeleton.
淋巴细胞和中性粒细胞的跨内皮迁移 (TEM) 与其可变形核移位内皮细胞骨架屏障的能力有关。核层粘连蛋白 A 是核层板的关键中间丝成分,在粒细胞生成过程中下调。当核层粘连蛋白 A 升高时,它会限制核通过刚性限制的挤压。为了确定白细胞核中低表达的核层粘连蛋白 A 是否对其通过内皮屏障的特殊挤压能力至关重要,我们在粒细胞样分化的 HL-60 细胞中过表达了这种蛋白质。核层粘连蛋白 A 表达增加 10 倍并不干扰这些粒细胞在固定的 CXCL1 上的趋化运动。此外,这些核层粘连蛋白 A 高的白细胞在大孔 Transwell 屏障中表现出正常的趋化性,向 CXCL1 运动,但在向相同的 CXCL1 梯度挤压时,通过 3 μm 孔的挤压能力很差。然而,令人惊讶的是,这些白细胞在剪切流下成功地完成了跨炎症内皮单层的 TEM,尽管在核挤压到其亚内皮伪足时略有延迟。相比之下,由于核层粘连蛋白 A 高的核不能转移到粒细胞的伪足中,因此,CXCR2 介导的粒细胞通过胶原蛋白 I 屏障的运动被核层粘连蛋白 A 过表达大大延迟。总的来说,我们的数据预测白细胞在核层板中保持低核层粘连蛋白 A 含量,以优化通过细胞外胶原蛋白屏障的挤压,但当穿过内皮细胞骨架提供的高度可适应的屏障时,它们可以耐受高核层粘连蛋白 A 含量。