Boneschansker Leo, Yan Jun, Wong Elisabeth, Briscoe David M, Irimia Daniel
Transplant Research Program and The Division of Nephrology, Department of Medicine, Boston Children's Hospital, Boston, MA 02115, and the Department of Pediatrics, Harvard Medical School, Boston, MA 02115.
BioMEMS Resource Center, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Shriners Hospitals for Children, Boston, MA 02129.
Nat Commun. 2014 Sep 3;5:4787. doi: 10.1038/ncomms5787.
Leukocyte migration into tissues is characteristic of inflammation. It is usually measured in vitro as the average displacement of populations of cells towards a chemokine gradient, not acknowledging other patterns of cell migration. Here, we designed and validated a microfluidic migration platform to simultaneously analyse four qualitative migration patterns: chemoattraction, -repulsion, -kinesis and -inhibition, using single-cell quantitative metrics of direction, speed, persistence and fraction of cells responding. We find that established chemokines, complement component 5a and IL-8 induce chemoattraction and repulsion in equal proportions, resulting in the dispersal of cells. These migration signatures are characterized by high persistence and speed and are independent of the chemokine dose or receptor expression. Furthermore, we find that twice as many T lymphocytes migrate away than towards stromal cell-derived factor 1 and their directional migration patterns are not persistent. Overall, our platform helps discover migratory signature responses and uncovers an avenue for precise characterization of leukocyte migration and therapeutic modulators.
白细胞迁移到组织中是炎症的特征。通常在体外通过细胞群体向趋化因子梯度的平均位移来测量,而不考虑细胞迁移的其他模式。在此,我们设计并验证了一种微流控迁移平台,以使用方向、速度、持续性和响应细胞分数的单细胞定量指标同时分析四种定性迁移模式:化学吸引、化学排斥、随机运动和抑制。我们发现,既定的趋化因子、补体成分5a和白细胞介素-8以相等比例诱导化学吸引和化学排斥,导致细胞分散。这些迁移特征以高持续性和速度为特点,且与趋化因子剂量或受体表达无关。此外,我们发现迁移离开的T淋巴细胞数量是迁移至基质细胞衍生因子1的两倍,且它们的定向迁移模式不具有持续性。总体而言,我们的平台有助于发现迁移特征反应,并揭示了精确表征白细胞迁移和治疗调节剂的途径。