title: Monoclonal antibodies directed against Leishmania secreted acid phosphatase and lipophosphoglycan. Partial characterization of private and public epitopes.
abstract: Leishmania promastigotes, the stage of the parasite characteristic for the sandfly vector, express an abundant glycoconjugate, called lipophosphoglycan, at their surface. Lipophosphoglycan consists of lysoalkyl-sn-glycerophosphoinositol linked to a phosphosaccharide core conserved in all species, which is connected to PO4-6Gal beta 1,4Man alpha 1 repeats with species-specific substitutions at the Gal residue; the repeats are capped by conserved and species-specific oligosaccharides. Most Leishmania species also secrete an acid phosphatase, which, in Leishmania mexicana, is a filamentous complex composed of a phosphorylated glycoprotein and non-covalently associated proteo-(high-molecular-mass)phosphoglycan. The secreted acid phosphatase complex was used as an antigen to derive a panel of monoclonal antibodies (mAbs). A total of 25 mAbs (17 novel and 8 previously described) were tested by different techniques for their specificity against lipophosphoglycan and secreted acid phosphatase from several Leishmania species. This comparison and the modification of the antigens by chemical or enzymic treatments allowed a classification of the mAbs into several groups. First, from 25 mAbs examined, 22 recognize lipophosphoglycan and the enzyme complex of L. mexicana; only three are specific for secreted acid phosphatase. Two of the latter group are also directed against carbohydrate structures, whereas the third mAb recognizes the 100-kDa polypeptide of the complex. The secreted acid-phosphatase-specific class detects antigen in the flagellar pocket of promastigotes while all anti-lipophosphoglycan mAbs bind to the cell surface. Second, all 15 anti-lipophosphoglycan mAbs investigated in detail appear to be directed against the phosphosaccharide repeats or the cap structure rather than the phosphosaccharide core. Two mAbs recognize terminal cap-structures containing Man alpha 1,2Man residues. Four antibodies are specific for L. mexicana and are probably directed against PO4-6[Glc beta 1,3]Gal beta 1,4Man alpha 1 repeats while six mAbs react with the unmodified repeats. Two antibodies specific for Leishmania major recognize Gal beta 1,3-substituted repeats unique for lipophosphoglycan from this species. Analysis by immunoblotting indicates that the high-molecular-mass proteo-phosphoglycan of L. mexicana secreted acid phosphatase carries epitopes for all anti-lipophosphoglycan mAbs suggesting the presence of capped phosphosaccharide repeats while the enzymically active glycoprotein subunit is modified by caps but probably not by repeats. In the case of Leishmania donovani secreted acid phosphatase, the enzymically active polypeptide may be directly modified by repeats. The mAbs are used to characterize changes in lipophosphoglycan structure, which occur in culture during the transition of promastigotes from the logarithmic to the stationary growth phase.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Max-Planck-Institut für Biologie, Tübingen, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Ilg
authors.forename: D
authors.initials: D
authors.lastname: Harbecke
authors.forename: M
authors.initials: M
authors.lastname: Wiese
authors.forename: P
authors.initials: P
authors.lastname: Overath
mesh_terms: D000135:Acid Phosphatase / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000913:Antibodies, Protozoan / Q000276:immunology*; D000918:Antibody Specificity; D001665:Binding Sites; D002240:Carbohydrate Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D006028:Glycosphingolipids / Q000276:immunology*; D006825:Hybridomas; D015151:Immunoblotting; D007891:Leishmania / Q000737:chemistry / Q000201:enzymology / Q000276:immunology*; D007894:Leishmania mexicana / Q000737:chemistry / Q000201:enzymology / Q000276:immunology; D008969:Molecular Sequence Data; D010766:Phosphorylation; D013045:Species Specificity
chemical_list: D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D000939:Epitopes; D006028:Glycosphingolipids; C008290:lipophosphonoglycan; D000135:Acid Phosphatase