Dynamic Adhesion of T Lymphocytes to Endothelial Cells Revealed by Atomic Force Microscopy

Abstract

The recruitment of T lymphocytes to lymphoid organs or sites of inflammation is a crucial step in adaptive immunity. These processes require endothelial activation and expression of adhesion molecules, including E- and P-selectins, intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1). However, the complete characterization of the adhesion strength and dynamics between lymphocytes and endothelial cells has been hampered by the lack of sensitive quantitative techniques. Here we report on the application of atomic force microscopy to characterize the interaction between individual pairs of living T lymphocytes (i.e., Jurkat cells) and human umbilical vein endothelial cells (HUVECs). The detachment of individual cell-cell conjugates was a complex process involving several step-like rupture events and the viscoelastic deformation of cells on the scale of several microns. Adhesion between Jurkat cells and activated endothelial cells increased with compression force and contact time, with the most dramatic changes occurring within the first half second of contact. After 0.25 sec of contact, E-selectin, ICAM-1, and VCAM-1 contributed to 18%, 39%, and 41% of total adhesion strength, respectively, suggesting that ICAM-1 and VCAM-1 contributed more than the selectins in supporting cell attachment.

Immune synapse formation determines interaction forces between T cells and antigen-presenting cells measured by atomic force microscopy

“…Benoit and coworkers pioneered the field of AFM based cell-cell adhesion measurements with Dictyostelium discoideum cells and found force values in the low nN regime for short contact times (11). In another study, Moy and coworkers examined interaction forces between human umbilical vein endothelial cells (HUVECs) and Jurkat T cells over a period of 10 s, yielding values in the range of 100 pN (13). Thus, so far, the majority of adhesion-related SCFS studies performed by AFM concentrated on short-time effects.…”

“…Cell-cell adhesion has been studied by micropipette aspiration techniques (9, 10) and atomic force microscopy (AFM) (11)(12)(13). The recent years have seen a significant increase of AFM-related studies in biological systems, and single-cell force spectroscopy (SCFS) by AFM has been established as an important tool for the study of cell adhesion (14).…”

2B4 Engagement Mediates Rapid LFA-1 and Actin-Dependent NK Cell Adhesion to Tumor Cells as Measured by Single Cell Force Spectroscopy

“…Most AFM approaches have focused on isolated receptor ligand interactions either using purified molecules or cells interacting with immobilized purified ligands (23,24). Other SCFS experiments have only concentrated on short-time effects, such as a 10-s contact between endothelial cells and Jurkat T cells (25). In this study, interaction forces of only 100 pN were reported.…”

Involvement of endothelial ephrin-B2 in adhesion and transmigration of EphB-receptor-expressing monocytes

“…Concanavalin A (ConA) functionalization of cantilevers was performed as described previously (Zhang et al, 2006). In brief, tipless Arrow TL1 cantilevers (Nanoworld, Neuchâtel, Switzerland) with a nominal spring constant of 0.03 N/m were used.…”