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Trypanosoma rhodesiense Bloodstream Trypomastigote and Culture Procyclic Cell Surface Carbohydrates 1, 2, 3
Author(s) -
JACKSON PETER R.,
DIGGS CARTER L.
Publication year - 1983
Publication title -
the journal of protozoology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 0022-3921
DOI - 10.1111/j.1550-7408.1983.tb05340.x
Subject(s) - agglutination (biology) , wheat germ agglutinin , lectin , soybean agglutinin , microbiology and biotechnology , agglutinin , biology , peanut agglutinin , biochemistry , horseradish peroxidase , concanavalin a , chemistry , antigen , enzyme , immunology , in vitro
Bloodstream trypomastigote and culture procyclic (insect midgut) forms of a cloned T. rhodesiense variant (WRATat 1) were tested for agglutination with the lectins concanavalin A (Con A), phytohemagglutinin P (PP), soybean agglutinin (SBA), fucose binding protein (FBP), wheat germ agglutinin (WGA), and castor bean lectin (RCA). Fluorescence‐microscopic localization of lectin binding to both formalin‐fixed trypomastigotes and red cells was determined with fluorescein isothiocyanate (FITC)‐conjugated Con A, SBA, FBP, WGA, RCA, PNA (peanut agglutinin), DBA (Dolichos bifloris) , and UEA (Ulex europaeus) lectins. Electron microscopic localization of lectin binding sites on bloodstream trypomastigotes was accomplished by the Con A‐horseradish peroxidase‐diaminobenzidine (HRP‐DAB) technique, and by a Con A‐biotin/avidin‐ferritin method. Trypomastigotes, isolated by centrifugation or filtration through DEAE‐cellulose or thawed after cryopreservation, were agglutinated by the lectins Con A and PP with agglutination strength scored as Con A < PP. No agglutination was observed in control preparations or with the lectins WGA, FBA or SBA. Red cells were agglutinated by all the lectins tested. Formalin‐fixed bloodstream trypomastigotes bound FITC‐Con A and FITC‐RCA but not FITC‐WGA, ‐SBA, ‐PNA, ‐UEA or ‐DBA lectins. All FITC‐labeled lectins bound to red cells. Con A receptors, visualized by Con A‐HRP‐DAB and Con A‐biotin/avidin‐ferritin techniques, were distributed uniformly on T. rhodesiense bloodstream forms. No lectin receptors were visualized on control preparations. Culture procyclics lacked a cell surface coat and were agglutinated by Con A and WGA but not RCA, SBA, PP and FBP. Procyclics were not agglutinated by lectins in the presence of competing sugar at 0.25 M. The expression of lectin binding cell surface saccharides of T. rhodesiense WRATat 1 is related to the parasite stage. Sugars resembling α‐D‐mannose are on the surface of bloodstream trypomastigotes and culture procyclics; n‐acetyl‐D‐galactosamine and D‐galactose residues are on bloodstream forms; and n‐acetyl‐D‐glucosamine‐like sugars are on procyclic stages.