Abstract
Gut-associated glycoproteins constitute a major group of the circulating excretory antigens produced by human Schistosoma species. The O-glycans of the relatively abundant circulating anodic antigen (CAA) from S. mansoni carry long stretches of unique →6(GlcAβ1→3)GalNAcβ1→ repeats. Specific anti-carbohydrate monoclonal antibodies (mAbs) are essential tools for the immunodiagnostic detection of CAA in the serum or urine of Schistosoma-infected subjects. In order to define the epitopes recognised by these anti-CAA mAbs, we screened a series of protein-coupled synthetic di- to pentasaccharide building blocks of the CAA polysaccharide for immunoreactivity, using ELISA and surface plasmon resonance spectroscopy. It was shown that anti-CAA IgM mAbs preferentially recognise →6(GlcAβ1→3)GalNAcβ1→ disaccharide units. Interestingly, no mouse anti-CAA mAbs of the IgG class were found that bind to the synthetic epitopes, although many of the IgG mAbs tested do recognise native CAA in a carbohydrate-dependent manner. In addition, both IgM and IgG class antibodies could be detected in human infection sera using the synthetic CAA fragments. These synthetic schistosome glycan epitopes and their matching set of specific mAbs are useful tools that further the development of diagnostic methods and are helpful in defining the immunological responses of the mammalian hosts to schistosome glycoconjugates.
Similar content being viewed by others
References
Berggren WL, Weller TH (1967) Immunoelectrophoretic demonstration of specific circulating antigen in animals infected with Schistosoma mansoni. Am J Trop Med Hyg 16:606–612
Bergwerff AA, van Dam GJ, Rotmans JP, Deelder AM, Kamerling JP, Vliegenthart JFG (1994) The immunologically reactive part of immunopurified circulating anodic antigen from Schistosoma mansoni is a threonine-linked polysaccharide consisting of →6)-[β-D-GlcpA-(1→3)]-β-D-GalpNAc-(1→ repeating units. J Biol Chem 269:31510–31517
Bos E, van der Doelen A, van Rooy N, Schuurs A (1981) 3,3′,5,5′-tetramethylbenzidine as an Ames test negative chromogen for horseradish peroxidase in enzyme immunoassay. J Immunoassays 2:187–196
Carlier Y, Bout D, Strecker G, Debray H, Capron A. (1980) Purification, immunochemical, and biological characterisation of the Schistosoma circulating M antigen. J Immunol 124:2442–2450
Cummings RD, Nyame AK (1999) Schistosome glycoconjugates. Biochim Biophys Acta 1455:363–374
De Jonge N, de Caluwé P, Hilberath GW, Krijger FW, Polderman AM, Deelder AM (1989) Circulating anodic antigen levels in serum before and after chemotherapy with praziquantel in Schistosoma mansoni. Trans R Soc Trop Med Hyg 83:368–372
Deelder AM, Kornelis D (1981) Immunodiagnosis of recently acquired Schistosoma mansoni infection. A comparison of various immunological techniques. Trop Geogr Med 33:36–41
Deelder AM, van Dalen DP, van Egmond JG (1978) Schistosoma mansoni: microfluorometric determination of circulating anodic antigen and antigen-antibody complexes in infected hamster serum. Exp Parasitol 44:216–224
Deelder AM, Kornelis D, van Marck EAE, Eveleigh PC, van Egmond JG (1980) Schistosoma mansoni: characterisation of two circulating polysaccharide antigens and the immunological response to these antigens in mouse, hamster, and human infections. Exp Parasitol 50:16–32
Deelder AM, de Jonge N, Boerman OC, Fillié YE, Hilberath GW, Rotmans JP, Gerritse MJ, Schut DWOA (1989) Sensitive determination of circulating anodic antigen in Schistosoma mansoni infected individuals by an enzyme-linked immunosorbent assay using monoclonal antibodies. Am J Trop Med Hyg 40:268–272
Deelder AM, van Dam GJ, Kornelis D, Fillié YE, van Zeyl RJM (1996) Schistosoma: analysis of monoclonal antibodies reactive with the circulating antigens CAA and CCA. Parasitology 112:21–35
Halkes KM, Vermeer HJ, Slaghek TM, van Hooft PAV, Loof A, Kamerling JP, Vliegenthart JFG (1998) Preparation of spacer-containing di-, tri-, and tetrasaccharide fragments of the circulating anodic antigen of Schistosoma mansoni for diagnostic purposes. Carbohydr Res 309:175–188
Hancock K, Tsang VCW (1986) Development and optimization of the FAST-ELISA for detecting antibodies to Schistosoma mansoni. J Immunol Methods 92:167–176
Hokke CH, Deelder AM (2001) Schistosome glycoconjugates in host-parasite interplay. Glycoconj J 18:573–587
MacKenzie CR, Hirama T, Deng S-J, Bundle DR, Narang SA, Young NM (1996) Analysis by surface plasmon resonance of the influence of valence on the ligand binding affinity and kinetics of an anti-carbohydrate antibody. J Biol Chem 271:1527–1533
Mushens RE, Scott ML (1990) A fast and efficient method for quantification of monoclonal antibodies in an ELISA using a novel incubation system. J Immunol Methods 131:83–89
Myszka DG (1999) Improving biosensor analysis. J Mol Recognit 12:1–6
Nash TE (1974) Localization of the circulating antigen within the gut of Schistosoma mansoni. Am J Trop Med Hyg 23:1085-1087
Nash TE, Nasir-ud-din, Jeanloz RW (1977) Further purification and characterisation of a circulating antigen in schistosomiasis. J Immunol 119:1627-1633
Nieba L, Krebber A, Plückthun A (1996) Competition BIAcore for measuring true affinities: large differences from values determined from binding kinetics. Anal Biochem 234:155-165
Nourel Din MS, Nibbeling R, Rotmans JP, Polderman AM, Krijger FW, Deelder AM (1994) Quantitative determination of circulating soluble egg antigen in urine and serum of Schistosoma mansoni-infected individuals using a combined two-site enzyme-linked immunosorbent assay. Am J Trop Med Hyg 50:585-94
Van Dam GJ, Seino J, Rotmans JP, Daha MR, Deelder AM (1993) Schistosoma mansoni circulating anodic antigen but not circulating cathodic antigen interacts with complement component C1q. Eur J Immunol 23:2807-2812
Van Lieshout L, Polderman AM, Deelder AM (2000) Immunodiagnosis of schistosomiasis by determination of the circulating antigens CAA and CCA, in particular in individuals with recent or light infections. Acta Trop 77:69-80
Van Remoortere A, Hokke CH, van Dam GJ, van Die I, Deelder AM, van den Eijnden DH (2000) Various stages of Schistosoma express Lewisx, LacdiNAc, GalNAcβ1-4(Fucα1-3)GlcNAc and GalNAcβ1-4(Fucα1-2Fucα1-3)GlcNAc carbohydrate epitopes: detection with monoclonal antibodies that are characterized by enzymatically synthesised neoglycoproteins. Glycobiology 86:410-413
Van Remoortere A, van Dam GJ, Hokke CH, van den Eijnden DH, van Die I, Deelder AM (2001a) Profiles of immunoglobulin M (IgM) and IgG antibodies against defined carbohydrate epitopes in sera of Schistosoma-infected individuals determined by surface plasmon resonance. Infect Immun 69:2396-2401
Van Remoortere A, Vermeer HJ, Ágoston K, Kerékgyártó J, van Roon AM, Wilson RA, Langermans JAM, van Die I, van den Eijnden DH, Vliegenthart JFG, Kamerling JP, Hokke CH, van Dam GJ, Deelder AM (2001b) Analysis by surface plasmon resonance of anti-carbohydrate responses to synthetic glycan epitopes in Pan troglodytes vaccinated with irradiated Schistosoma mansoni cercariae. Glycoconj J 18: Abstract C16.11
Vermeer HJ, Halkes KM, Kamerling JP, Vliegenthart JFG (2000) Synthesis and conjugation of oligosaccharide fragments related to the immunologically reactive part of the circulating anodic antigen of the parasite Schistosoma mansoni. J Chem Soc, Perkin Trans 1: 2249-2263.
Acknowledgements
The authors wish to thank Dr. S. Haseley for critical reading of the manuscript, and Ms. A. van Remoortere for helpful discussions. The technical assistance of Ms. D. Kornelis, Mr. R.J.M. van Zeyl, and Ms. S. Ferreira is highly appreciated.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Vermeer, H.J., van Dam, G.J., Halkes, K.M. et al. Immunodiagnostically applicable monoclonal antibodies to the circulating anodic antigen of Schistosoma mansoni bind to small, defined oligosaccharide epitopes. Parasitol Res 90, 330–336 (2003). https://doi.org/10.1007/s00436-003-0860-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-003-0860-3