Leishmania tropica: characterization of a lipophosphoglycan-like antigen recognized by species-specific monoclonal antibodies.
Species-specific monoclonal antibodies to Leishmania tropica, T11 and T13-15, recognize membranal and secreted antigens. The membrane form of the antigen migrates on sodium dodecyl sulfate-polyacrylamide gels with a diffuse molecular weight from 15 to 50 kDa and can be labeled with palmitic acid, myoinositol, galactose, glucosamine, and inorganic phosphate. Both phosphate and sugar-labeled material were isolated from metabolically labeled promastigotes by affinity chromatography on antibodies coupled to Sepharose 4B. No binding to Ricinus communis agglutinin was observed. This material behaves like lipophosphoglycans from other Leishmania but contains unique species-specific epitopes. It is susceptible to cleavage by phospholipase C and after digestion no longer partitions into the detergent phase following a Triton X-114 extraction. All four monoclonal antibodies appear to recognize a carbohydrate epitope on the lipophosphoglycan since periodate treatment of this material bound to nitrocellulose essentially eliminated antibody binding. In addition, T15 binding could be blocked by 5 mM mannose-6-PO4 and fructose-1- or 6-PO4, but not by mannose, glucose, fructose, or the additional PO4 derivatives examined. The antibodies recognize a similar but not identical epitope, as demonstrated by a competitive radioimmunoassay using 125I-labeled T11, T13, and T15. Expression of surface antigen is elevated during the promastigote stationary phase.
Department of Biophysics, Weizmann Institute of Science, Rehovot, Israel.
C L
CL
Jaffe
M L
ML
Pérez
R
R
Sarfstein
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000953:Antigens, Protozoan / Q000032:analysis* / Q000276:immunology; D001667:Binding, Competitive; D002241:Carbohydrates / Q000494:pharmacology; D002469:Cell Separation; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis / Q000276:immunology; D005434:Flow Cytometry; D006028:Glycosphingolipids / Q000032:analysis* / Q000276:immunology; D007895:Leishmania tropica / Q000276:immunology*; D011233:Precipitin Tests; D011863:Radioimmunoassay; D013045:Species Specificity; D010738:Type C Phospholipases / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D002241:Carbohydrates; D000939:Epitopes; D006028:Glycosphingolipids; C008290:lipophosphonoglycan; D010738:Type C Phospholipases