Phagocytosis of liposomes by macrophages: intracellular fate of liposomal malaria antigen.
Liposomes containing a synthetic recombinant protein were phagocytosed by macrophages, and the internalized protein was recycled to the cell surfaces where it was detected by enzyme-linked immunosorbent assay. The transit time of the liposome-encapsulated protein from initial phagocytosis of liposomes to appearance of protein on the surfaces of macrophages was determined by pulse-chase experiments. The macrophages were pulsed with liposomes containing protein and chased with empty liposomes, and vice versa. The amount and rate of protein antigen expression at the cell surfaces depended on the quantity of encapsulated protein ingested by the macrophages. Although liposomes were rapidly taken up by macrophages, the liposome-encapsulated protein was antigenically expressed for a prolonged period (at least 24 h) on the cell surface. Liposomes were visualized inside vacuoles in the macrophages by immunogold electron microscopy. The liposomes accumulated along the peripheries of the vacuoles and many of them apparently remained intact for a long time (greater than 6 h). However, nonliposomal free protein was also detected in the cytoplasm surrounding these vacuoles, and it was concluded that the free protein in the cytoplasm was probably en route to the macrophage surface. Exposure of the cells to ammonium chloride did not inhibit the appearance of liposomal antigenic epitopes on the cell surface, and this suggests that expression of the liposomal antigenic epitopes at the surface was not a pH-sensitive phenomenon. There was no significant effect of a liposomal adjuvant, lipid A, on the rate or extent of surface expression of the liposomal protein.
Verma
J N
JN
Department of Membrane Biochemistry, Walter Reed Army Institute of Research, Washington, DC 20307-5100.
Wassef
N M
NM
Wirtz
R A
RA
Atkinson
C T
CT
Aikawa
M
M
Loomis
L D
LD
Alving
C R
CR
D000595:Amino Acid Sequence; D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D000953:Antigens, Protozoan / Q000378:metabolism*; D000956:Antigens, Viral / Q000378:metabolism; D001854:Bone Marrow Cells; D004797:Enzyme-Linked Immunosorbent Assay; D007700:Kinetics; D008050:Lipid A / Q000494:pharmacology; D008081:Liposomes / Q000276:immunology* / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D051379:Mice; D008809:Mice, Inbred C3H; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D010587:Phagocytosis; D010963:Plasmodium falciparum / Q000276:immunology*; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D000953:Antigens, Protozoan; D000956:Antigens, Viral; D008050:Lipid A; D008081:Liposomes; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; C041595:circumsporozoite protein, Protozoan; D000643:Ammonium Chloride