Mapping fucosylated epitopes on glycoproteins and glycolipids of Schistosoma mansoni cercariae, adult worms and eggs.
The developmental expression of the antigenic fucosylated glycan motifs Fucalpha1-3GalNAcbeta1-4GlcNAc (F-LDN), Fucalpha1-3GalNAcbeta1-4(Fucalpha1-3)GlcNAc (F-LDN-F), GalNAcbeta1-4(Fucalpha1-3)GlcNAc (LDN-F), Galbeta1-4(Fucalpha1-3)GlcNAc (Lewis X), and GalNAcbeta1-4(Fucalpha1-2Fucalpha1-3)GlcNAc (LDN-DF) in Schistosoma mansoni cercariae, adult worms and eggs, was surveyed using previously defined anti-carbohydrate monoclonal antibodies (mAbs). Lewis X was found both on glycolipids and glycoproteins, yet with completely different expression patterns during the life-cycle: on glycolipids, Lewis X was mainly found in the cercarial stage, while protein-conjugated Lewis X was mainly present in the egg stage. Also protein-conjugated LDN-F and LDN-DF were most highly expressed in the egg-stage. On glycolipids LDN-DF was found in all three examined stages, whereas LDN-F containing glycolipids were restricted to adult worms and eggs. The motifs F-LDN and F-LDN-F were found both on glycoproteins and glycolipids of the cercarial and egg stage, while in the adult stage, they appeared to occur predominantly on glycolipids. Immunofluorescence assays (IFA) showed that these F-LDN and F-LDN-F containing glycolipids were localized in a yet undefined duct or excretory system of adult worms. Murine infection serum showed major reactivity with this adult worm duct-system, which could be fully inhibited by pre-incubation with keyhole limpet haemocyanin (KLH). Clearly, the use of defined mAbs provides a quick and convenient way to map expression profiles of carbohydrate epitopes.
Robijn
M L M
ML
Department of Parasitology, Centre of Infectious Diseases, Leiden University Medical Centre, 2300 RC Leiden, The Netherlands.
Wuhrer
M
M
Kornelis
D
D
Deelder
A M
AM
Geyer
R
R
Hokke
C H
CH
D020816:Amino Acid Motifs; D000818:Animals; D002240:Carbohydrate Sequence; D018604:Epitope Mapping; D005643:Fucose / Q000032:analysis*; D018507:Gene Expression Regulation, Developmental; D006017:Glycolipids / Q000737:chemistry / Q000276:immunology*; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology*; D015801:Helminth Proteins / Q000276:immunology; D008018:Life Cycle Stages / Q000276:immunology; D008969:Molecular Sequence Data; D010063:Ovum / Q000276:immunology; D012550:Schistosoma mansoni / Q000254:growth & development* / Q000276:immunology*
D006017:Glycolipids; D006023:Glycoproteins; D015801:Helminth Proteins; D005643:Fucose