Interaction of cationic colloids at the surface of J774 cells: a kinetic analysis.
We have characterized the binding of multilamellar colloids to J774 cells. Cationic colloids were shown to bind much more efficiently than neutral ones. Particle uptake by cells was followed by flow cytometry and fluorescence microscopy. Analysis of the kinetics of uptake of cationic particles indicated that binding on the cell surface occurred with two characteristic times. Analysis of the dissociation properties allowed discriminating between several alternative models for adsorption and led us to propose a mechanism that involved two independent classes of binding sites on the cell surface. One class of sites appeared to be governed by a classic mass action law describing a binding equilibrium. The other sites were populated irreversibly by particles made of 10% cationic lipids. This was observed in the absence of endocytosis, under conditions where both the equilibrium and the irreversible binding occurred at the cell surface. We determined the rate constants for the different steps. We found that the reversible association occurred with a characteristic time of the order of tens of seconds, whereas the irreversible binding took a hundred times longer. The presence of serum proteins in the incubation medium did not drastically affect the final uptake of the particles. In contrast, the capture of the particles by cells significantly dropped when the fraction of positively charged lipids contained in the colloids was decreased from 10% to 5%. Finally, the results will be discussed within a comprehensive model where cationic particles find labile binding sites in the volume of the pericellular network (glycocalyx and extracellular matrix) and less-accessible irreversible binding sites at the cell membrane itself.
Chenevier
P
P
Centre de Recherche Paul Pascal, CNRS, 33600 Pessac, France.
Veyret
B
B
Roux
D
D
Henry-Toulmé
N
N
D000818:Animals; D001665:Binding Sites; D002412:Cations / Q000493:pharmacokinetics; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology*; D003102:Colloids / Q000493:pharmacokinetics*; D005434:Flow Cytometry / Q000379:methods; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008856:Microscopy, Fluorescence / Q000379:methods; D008954:Models, Biological; D008962:Models, Theoretical
D002412:Cations; D003102:Colloids