Energy of adhesion of human T cells to adsorption layers of monoclonal antibodies measured by a film trapping technique.
A novel method for studying the interaction of biological cells with interfaces (e.g., adsorption monolayers of antibodies) is developed. The method is called the film trapping technique because the cell is trapped within an aqueous film of equilibrium thickness smaller than the cell diameter. A liquid film of uneven thickness is formed around the trapped cell. When observed in reflected monochromatic light, this film exhibits an interference pattern of concentric bright and dark fringes. From the radii of the fringes one can restore the shape of interfaces and the cell. Furthermore, one can calculate the adhesive energy between the cell membrane and the aqueous film surface (which is covered by a layer of adsorbed proteins and/or specific ligands), as well as the disjoining pressure, representing the force of interaction per unit area of the latter film. The method is applied to two human T cell lines: Jurkat and its T cell receptor negative (TCR-) derivative. The interaction of these cells with monolayers of three different monoclonal antibodies adsorbed at a water-air interface is studied. The results show that the adhesive energy is considerable (above 0.5 mJ/m2) when the adsorption monolayer contains antibodies acting as specific ligands for the receptors expressed on the cell surface. In contrast, the adhesive energy is close to zero in the absence of such a specific ligand-receptor interaction. In principle, the method can be applied to the study of the interaction of a variety of biological cells (B cells, natural killer cells, red blood cells, etc.) with adsorption monolayers of various biologically active molecules. In particular, film trapping provides a tool for the gentle micromanipulation of cells and for monitoring of processes (say the activation of a T lymphocyte) occurring at the single-cell level.
Ivanov
I B
IB
Laboratory of Thermodynamics and Physico-chemical Hydrodynamics, Faculty of Chemistry, University of Sofia, 1126 Sofia, Bulgaria.
Hadjiiski
A
A
Denkov
N D
ND
Gurkov
T D
TD
Kralchevsky
P A
PA
Koyasu
S
S
D000327:Adsorption; D000911:Antibodies, Monoclonal; D001703:Biophysics / Q000295:instrumentation / Q000379:methods*; D002448:Cell Adhesion / Q000502:physiology*; D002462:Cell Membrane / Q000276:immunology / Q000502:physiology / Q000648:ultrastructure; D006801:Humans; D066298:In Vitro Techniques; D007368:Interferometry; D019169:Jurkat Cells; D008027:Light; D008954:Models, Biological; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology / Q000502:physiology* / Q000648:ultrastructure; D013816:Thermodynamics
D000911:Antibodies, Monoclonal; D011948:Receptors, Antigen, T-Cell