Surface carbohydrates of procyclic forms of African trypanosomes studied using fluorescence activated cell sorter analysis and agglutination with lectins.
Living culture form procyclics of Trypanosoma brucei brucei, T.b. rhodesiense, T.b. gambiense, T. congolense and T. simiae were tested for binding of eight different lectins. The binding of fluorescein isothiocyanate (FITC)-conjugated lectins was measured using a fluorescence activated cell sorter (FACS) and by agglutination with unlabelled lectins. Five of the lectins failed to bind to any of the procyclic organisms in both tests. All parasites bound concanavalin A (Con A) and all T.b. brucei, T.b. rhodesiense and T. congolense procyclics bound Ricinus communis agglutinin 120 (RCA) and wheat germ agglutinin (WGA). Trypanosoma b. gambiense procyclics failed to bind RCA and thus could be easily discriminated from other subspecies of T. brucei. Similarly, T. simiae did not bind WGA, unlike T. congolense, the other species of the genus Nannomonas. All positive reactions were inhibited by 0.2 M concentrations of the relevant sugars. The results indicate that all species and subspecies of the procyclic culture forms tested have surface-exposed structures resembling alpha-D-mannose moieties and that T.b. brucei, T.b. rhodesiense and T. congolense have surface-exposed molecules resembling D-galactose and N-acetyl D-glucosamine (or sialic acid) moieties. Molecules resembling D-galactose and N-acetyl D-glucosamine residues are absent or inaccessible in T.b. gambiense and T. simiae respectively. A group of T. congolense clones of parasite stocks isolated at Kilifi on the Kenyan coast showed quantitatively different binding of RCA when compared to the other T. congolense clones tested indicating that these organisms differ in surface carbohydrate structure.
D000371:Agglutination; D000818:Animals; D050260:Carbohydrate Metabolism; D002469:Cell Separation; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins / Q000378:metabolism; D037102:Lectins / Q000378:metabolism; D013861:Thiocyanates / Q000378:metabolism; D014345:Trypanosoma / Q000378:metabolism*; D014347:Trypanosoma brucei gambiense / Q000378:metabolism; D014348:Trypanosoma congolense / Q000378:metabolism
D005452:Fluoresceins; D037102:Lectins; D013861:Thiocyanates; D016650:Fluorescein-5-isothiocyanate