Various stages of schistosoma express Lewis(x), LacdiNAc, GalNAcbeta1-4 (Fucalpha1-3)GlcNAc and GalNAcbeta1-4(Fucalpha1-2Fucalpha1-3)GlcNAc carbohydrate epitopes: detection with monoclonal antibodies that are characterized by enzymatically synthesized neoglycoproteins.
We report here that fucosylated epitopes such as Lewis(x), LacdiNAc, fucosylated LacdiNAc (LDN-F) and GalNAcbeta1-4(Fucalpha1-2Fucalpha1-3)GlcNAc (LDN-DF) are expressed by schistosomes throughout their life cycle. These four epitopes were enzymatically synthesized and coupled to bovine serum albumin to yield neoglycoproteins. Subsequently these neoglycoproteins were used to probe a panel of 188 monoclonal antibodies obtained from infected or immunized mice, in ELISA and surface plasmon resonance analysis. Of these antibodies, 25 recognized one of the fucosylated structures synthesized, indicating that these structures are immunogenic during infection. The MAbs identified could be subdivided in four different groups based on the recognition of either the Lewis(x)-, the LacdiNAc-, the LDN-DF-, or both the LDN-F- and LDN-DF epitope. These monoclonal antibodies were then used to investigate the localization of the fucosylated epitopes in various stages of Schistosoma mansoni using indirect immunofluorescence. Lewis(x)epitopes were mainly found in the gut and on the tegument of adult worms, on egg shells, and on the oral sucker of cercariae. The LacdiNAc epitope was expressed on the tegument of adult worms, on miracidia, and on the oral sucker of cercariae. In contrast, LDN-DF epitopes were mainly present in the excretory system of adult worms, on miracidia and on whole cercariae. These also stained positive with the LDN-F/LDN-DF epitope antibodies, while whole parenchyma reacted characteristically only with the latter antibodies. The identification of different carbohydrate structures in various stages of schistosomes may lead to a better understanding of the function of glycans in the immune response during infection.
van Remoortere
A
A
Department of Medical Chemistry, Vrije Universiteit, Amsterdam, the Netherlands.
Hokke
C H
CH
van Dam
G J
GJ
van Die
I
I
Deelder
A M
AM
van den Eijnden
D H
DH
D000818:Animals; D000911:Antibodies, Monoclonal; D006224:Cricetinae; D004187:Disaccharides / Q000032:analysis* / Q000378:metabolism; D000939:Epitopes / Q000032:analysis*; D019084:Fluorescent Antibody Technique, Indirect; D005643:Fucose / Q000378:metabolism*; D006023:Glycoproteins / Q000032:analysis / Q000138:chemical synthesis / Q000276:immunology; D007785:Lactose / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D016256:Lewis X Antigen / Q000032:analysis*; D051379:Mice; D012550:Schistosoma mansoni / Q000254:growth & development* / Q000276:immunology*; D012555:Schistosomiasis mansoni / Q000276:immunology; D020349:Surface Plasmon Resonance; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D004187:Disaccharides; D000939:Epitopes; D006023:Glycoproteins; D016256:Lewis X Antigen; C093701:N-acetylgalactosaminyl-1-4-N-acetylglucosamine; D005643:Fucose; D007785:Lactose