Enhanced binding of phosphatidylserine-containing lipid vesicle targets to RAW264 macrophages.
Phosphatidylserine was found to significantly enhance the binding of phospholipid vesicles to RAW264 macrophages. We have measured the kinetics of non-specific uptake of unilamellar vesicles as a function of phosphatidylserine concentration in these model target membranes. Dimyristoylphosphatidylcholine was the principle component of these phospholipid vesicles. In most experiments, radiolabeled phospholipid and 1 mol% each of both a fluorescent phospholipid and a hapten-containing lipid headgroup were utilized. In the presence of specific anti-hapten antibody phosphatidylserine-containing vesicles are rapidly taken up via phagocytosis. The antibody-independent non-specific uptake of phosphatidylserine-free vesicles was low, as previously reported. However, the presence of 5 mol% phosphatidylserine dramatically enhanced the uptake of phospholipid vesicles by macrophages. This uptake was shown to be principally due to binding to the macrophage surface. Incubation of macrophages in the presence of sodium azide or at 4 degrees C, conditions which are known to inhibit phagocytosis, do not influence the uptake of the lipid vesicles. Fluorescence video-intensification microscopy was used to observe the interaction of carboxyfluorescein-loaded vesicles with macrophages. Fluorescence could not be observed when using phosphatidylserine-free vesicles. However, phosphatidylserine-containing vesicles can be observed bound to the cell periphery. Intracellular fluorescence could not be observed. The binding of phosphatidylserine-containing vesicles was enhanced roughly four-fold over phosphatidylserine because the effect could not be observed with membranes containing 1 mol% or 2.5 mol % phosphatidylserine. In addition, the binding enhancement required the presence of divalent cations in the incubation medium.
D000818:Animals; D000906:Antibodies; D002118:Calcium / Q000502:physiology; D002460:Cell Line; D004140:Dinitrophenols / Q000276:immunology; D008081:Liposomes; D008264:Macrophages / Q000502:physiology*; D008563:Membrane Lipids / Q000502:physiology; D051379:Mice; D010587:Phagocytosis; D010718:Phosphatidylserines / Q000502:physiology
D000906:Antibodies; D004140:Dinitrophenols; D008081:Liposomes; D008563:Membrane Lipids; D010718:Phosphatidylserines; D002118:Calcium