Single-cell adhesive profiling in an optofluidic device elucidates CD8+ T lymphocyte phenotypes in inflamed vasculature-like microenvironments

Camila P Camargo; Yunus Alapan; Abir K Muhuri; Samuel N Lucas; Susan N Thomas

doi:10.1016/j.crmeth.2024.100743

Single-cell adhesive profiling in an optofluidic device elucidates CD8⁺ T lymphocyte phenotypes in inflamed vasculature-like microenvironments

Cell Rep Methods. 2024 Apr 22;4(4):100743. doi: 10.1016/j.crmeth.2024.100743. Epub 2024 Mar 29.

Authors

Camila P Camargo¹, Yunus Alapan¹, Abir K Muhuri¹, Samuel N Lucas², Susan N Thomas³

Affiliations

¹ George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta 30332, GA, USA; Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta 30332, GA, USA.
² Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta 30332, GA, USA; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta 30332, GA, USA.
³ George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta 30332, GA, USA; Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta 30332, GA, USA; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta 30332, GA, USA; Winship Cancer Institute, Emory University, Atlanta 30322, GA, USA. Electronic address: susan.thomas@gatech.edu.

Abstract

Tissue infiltration by circulating leukocytes occurs via adhesive interactions with the local vasculature, but how the adhesive quality of circulating cells guides the homing of specific phenotypes to different vascular microenvironments remains undefined. We developed an optofluidic system enabling fluorescent labeling of photoactivatable cells based on their adhesive rolling velocity in an inflamed vasculature-mimicking microfluidic device under physiological fluid flow. In so doing, single-cell level multidimensional profiling of cellular characteristics could be characterized and related to the associated adhesive phenotype. When applied to CD8⁺ T cells, ligand/receptor expression profiles and subtypes associated with adhesion were revealed, providing insight into inflamed tissue infiltration capabilities of specific CD8⁺ T lymphocyte subsets and how local vascular microenvironmental features may regulate the quality of cellular infiltration. This methodology facilitates rapid screening of cell populations for enhanced homing capabilities under defined biochemical and biophysical microenvironments, relevant to leukocyte homing modulation in multiple pathologies.

Keywords: CP: biotechnology; CP: immunology; cell adhesion; homing; inflamed vasculature; lymphocyte; microfluidic system; photoactivation; selectins; single-cell.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Animals
CD8-Positive T-Lymphocytes* / immunology
CD8-Positive T-Lymphocytes* / metabolism
Cell Adhesion*
Cellular Microenvironment / immunology
Humans
Inflammation / immunology
Inflammation / pathology
Lab-On-A-Chip Devices
Mice
Mice, Inbred C57BL
Phenotype*
Single-Cell Analysis* / methods