Cardiff Chronic Lymphocytic Leukemia Research Group, Institute of Cancer & Genetics, School of Medicine, Cardiff University, Heath Park, Cardiff, United Kingdom;
Department of Haematology, King's College London, London, United Kingdom; and.
Blood. 2014 Jun 5;123(23):3607-17. doi: 10.1182/blood-2013-12-544569. Epub 2014 Mar 17.
There is growing evidence that lymphocyte trafficking contributes to the clinical course of chronic lymphocytic leukemia (CLL), but to date, only static in vitro cultures have been used to study these phenomena. To address this lack of data, we have developed a dynamic in vitro model in which CLL cells experience shear forces equivalent to those in capillary beds and are made to flow through capillary-like hollow fibers lined with endothelial cells. CLL cells treated in this way increased their expression of CD62L and CXCR4 (both P < .0001) and of CD49d and CD5 (both P = .003) directly as a result of the shear force. Furthermore, CLL cells migrated through the endothelium into the "extravascular" space (mean migration, 1.37% ± 2.14%; n = 21). Migrated CLL cells had significantly higher expression of CD49d (P = .02), matrix metallopeptidase-9 (P = .004), CD38 (P = .009), CD80 (P = .04), and CD69 (P = .04) compared with CLL cells that remained in the circulation. The degree of migration observed strongly correlated with CD49d expression (r(2), 0.47; P = .01), and treatment with the CD49d-blocking antibody natalizumab resulted in significantly decreased migration (P = .01). Taken together, our data provide evidence for a novel, dynamic, and tractable in vitro model of lymphocyte migration and confirm that CD49d is a critical regulator of this process in CLL.
越来越多的证据表明，淋巴细胞的迁移有助于慢性淋巴细胞白血病（CLL）的临床进程，但迄今为止，仅使用静态的体外培养来研究这些现象。为了解决这一数据缺乏的问题，我们开发了一种动态的体外模型，在该模型中，CLL 细胞经历与毛细血管床相同的剪切力，并通过内皮细胞排列的毛细血管样中空纤维流动。用这种方式处理的 CLL 细胞直接增加了其 CD62L 和 CXCR4 的表达（均 P <.0001）以及 CD49d 和 CD5 的表达（均 P =.003），这是由于剪切力的作用。此外，CLL 细胞通过内皮细胞迁移到“血管外”空间（平均迁移率为 1.37% ± 2.14%；n = 21）。迁移的 CLL 细胞显著更高地表达 CD49d（P =.02）、基质金属蛋白酶-9（P =.004）、CD38（P =.009）、CD80（P =.04）和 CD69（P =.04），与留在循环中的 CLL 细胞相比。观察到的迁移程度与 CD49d 表达强烈相关（r(2)，0.47；P =.01），并用 CD49d 阻断抗体那他珠单抗治疗导致迁移显著减少（P =.01）。总之，我们的数据提供了一种新型的、动态的、可处理的淋巴细胞迁移体外模型的证据，并证实 CD49d 是 CLL 中这一过程的关键调节因子。
