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A Method of Introducting Lipid-Conjugated Antigens into the Surface of Schistosomula
Introduction of synthetic antigens into the surface of schistosomula of Schistosoma mansoni was achieved by brief incubation of the worms with liposomes carrying the lipid-bound antigens in their bilayers. Three-hour-old schistosomula were surface-labeled with lipid-conjugated dinitrophenyl (DNP) groups by using liposomes made of egg lecithin-N-dinitrophenil-ε-aminocaproyl-phosphatidylethanolamine (5:1). The DNP groups incorporated in this way could be detected for more than 21 hours in vitro by using rabbit anti-DNP antibodies stained with fluorescein isothiocyanate-conjugated goat anti-rabbit IgG. Immunofluorescence microscopy showed the lipid antigen to be uniformly distributed over the entire surface of the worms. Electron microscope studies, performed with purified rabbit anti-DNP antibodies followed by ferritin-conjugated goat anti-rabbit IgG, showed that the DNP groups were evenly and densely distributed over the entire outer membrane of the schistosomula, including spines. The distance between the ferritin molecules and the parasite's surface was 24 ± 5 nm, indicating that the lipid antigen had been incorporated into the outer membrane of the schistosomula.