Lipid analyses of isolated surface membranes of Leishmania donovani promastigotes.

Constituent lipids of surface membranes (SM) isolated from Leishmania donovani promastigotes were analyzed and compared with those obtained from whole cells and an isolated kinetoplast-mitochondrion fraction (KM). On a dry weight basis, the total extractable lipids constituted approximately 47%, 12% and 24% of the SM, cells and KM, respectively. The total lipids of SM, cells and KM all were composed of approximately 70% phospholipids (PL), 20-25% neutral lipids and 5-10% glycolipids. Sterols and diglycerides composed 60% and 30%, respectively, of the various neutral lipid fractions. Several mannose- and galactose-containing glycolipids were fractionated but not identified. The glycolipid fractions from cells and SM had demonstrable antigenic activities with rabbit anti-SM sera. Striking quantitative differences were apparent between the PL profiles of the 3 cellular components examined. The PL of SM, whole cells and KM, respectively, were composed of: 15%, 51% and 24% phosphatidylcholine; 37%, 13% and 11% phosphatidylethanolamine (PE); 18%, 10% and 14% phosphatidylinositol; 10%, 1% and 4% phosphatidylserine and traces of cardiolipin, phosphatidylglycerol and phosphatidic acid. An unknown PL containing sphingosine, choline and vicinal hydroxyl groups but no free amino moieties made up approximately 19% of the PL of SM and whole cells, but it constituted approximately 27% of the PL of KM. The PL side chain constituents of whole cells and SM were composed mainly of longchain fatty acids (C18-20). Further, over 50% of the PE of SM was in the alkyl and alK-1-enyl ether forms. These SM properties might contribute to the organism's resistance to digestion in the hydrolytic environs of both its insect vector and mammalian hosts.

In vivo and in vitro localization of Leishmania within macrophage phagolysosomes: use of colloidal gold as a lysosomal label.

The interaction of Leishmania with lysosomes within macrophages in vivo has been investigated. Lysosomes labeled with colloidal gold in vivo fused with phagocytic vacuoles containing Leishmania amastigotes within the macrophages of infected footpad tissue of BALB/c mice. This localization of Leishmania within macrophage phagolysosomes in vivo is the first confirmation for any obligate intracellulaire protozoon that parasite-lysosome interactions in vitro occur in vivo.