Microfluidic mazes to characterize T-cell exploration patterns following activation in vitro.
The migration of T-cell subsets within peripheral tissues is characteristic of inflammation and immunoregulation. In general, the lymphocyte migratory response is assumed directional and guided by local gradients of chemoattractants and/or chemorepellents. However, little is known about how cells explore their tissue environment, and whether lymphocyte activation may influence speed and exploratory patterns of migration. To probe migration patterns by T-cells we designed a microfluidic maze device that replicates critical features of a tissue-like microenvironment. We quantified the migration patterns of unstimulated and mitogen-activated human T-cells at single cell resolution and found significant differences in exploration within microfluidic mazes. While unstimulated lymphocytes migrated in a directed manner, activated T-cells migrated through large areas of the mazes in an exploratory pattern in response to the chemoattractants RANTES (CCL5) and IP-10 (CXCL10). The analysis of migration enabled by the microfluidic devices help develop new methods for determining how human circulating T-cells function in vivo to seek out antigens in health and disease states.
BioMEMS Resource Center, Department of Surgery, Massachusetts General Hospital, Boston, MA 02129, USA and Transplant Research Program, Boston Children's Hospital, Boston, MA 02115, USA. David.Briscoe@childrens.harvard.edu.
Namrata G
NG
Jain
BioMEMS Resource Center, Department of Surgery, Massachusetts General Hospital, Boston, MA 02129, USA.
Elisabeth A
EA
Wong
BioMEMS Resource Center, Department of Surgery, Massachusetts General Hospital, Boston, MA 02129, USA.
Alexander J
AJ
Aranyosi
BioMEMS Resource Center, Department of Surgery, Massachusetts General Hospital, Boston, MA 02129, USA and Transplant Research Program, Boston Children's Hospital, Boston, MA 02115, USA. David.Briscoe@childrens.harvard.edu.
Leo
L
Boneschansker
BioMEMS Resource Center, Department of Surgery, Massachusetts General Hospital, Boston, MA 02129, USA and Division of Nephrology, Department of Medicine, Boston Children's Hospital, 300 Longwood Ave, MA 02139, USA.
James F
JF
Markmann
Transplant Research Program, Boston Children's Hospital, Boston, MA 02115, USA. David.Briscoe@childrens.harvard.edu and Division of Nephrology, Department of Medicine, Boston Children's Hospital, 300 Longwood Ave, MA 02139, USA.
David M
DM
Briscoe
BioMEMS Resource Center, Department of Surgery, Massachusetts General Hospital, Boston, MA 02129, USA and Harvard Medical School, Boston, MA 02129, USA. dirimia@hms.harvard.edu.
Daniel
D
Irimia
D000893:Anti-Inflammatory Agents / Q000737:chemistry; D000941:Antigens / Q000737:chemistry; D002478:Cells, Cultured; D018946:Chemokine CCL5 / Q000378:metabolism; D054357:Chemokine CXCL10 / Q000378:metabolism; D002634:Chemotaxis, Leukocyte; D064368:Healthy Volunteers; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008213:Lymphocyte Activation; D046210:Microfluidic Analytical Techniques; D044085:Microfluidics; D013601:T-Lymphocytes / Q000166:cytology*
D000893:Anti-Inflammatory Agents; D000941:Antigens; C483024:CCL5 protein, human; C114753:CXCL10 protein, human; D018946:Chemokine CCL5; D054357:Chemokine CXCL10