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Biochemical Journal logoLink to Biochemical Journal
. 2000 Jun 1;348(Pt 2):417–423.

Phosphocholine-containing, zwitterionic glycosphingolipids of adult Onchocerca volvulus as highly conserved antigenic structures of parasitic nematodes.

M Wuhrer 1, S Rickhoff 1, R D Dennis 1, G Lochnit 1, P T Soboslay 1, S Baumeister 1, R Geyer 1
PMCID: PMC1221081  PMID: 10816437

Abstract

Human Onchocerca volvulus infection sera were found to recognize zwitterionic glycolipids of O. volvulus and to cross-react with those of other parasitic nematodes (Ascaris suum, Setaria digitata and Litomosoides sigmodontis). By the use of an epitope-specific monoclonal antibody, zwitterionic glycolipids of all these nematode species were observed to contain the antigenic determinant phosphocholine. A hyperimmune serum specific for arthro-series glycolipid structures reacted with the various neutral glycolipids of all these nematodes, which demonstrated that their oligosaccharide moieties belonged to the arthro-series of protostomial glycolipids. These results indicated that arthro-series glycosphingolipids carrying, in part, phosphocholine substituents, represent highly conserved, antigenic glycolipid markers of parasitic nematodes. Three glycolipid components of the O. volvulus zwitterionic fraction were structurally characterized by matrix-assisted laser-desorption/ionization time-of-flight MS, methylation analysis and exoglycosidase treatment. Their chemical structures were elucidated to be phosphocholine-6GlcNAc(beta1-3)Man(beta1-4)Glc(1-1)ceramide, GalNAc(beta1-4)[phosphocholine-6]GlcNAc(beta1-3)Man(beta1-4)Glc(1-1) ceramide and Gal(alpha1-3)GalNAc(beta1-4)[phosphocholine-6]GlcNAc(beta1-3)Man(beta 1-4)Glc(1-1)ceramide for the zwitterionic ceramide tri-, tetra- and penta-hexosides respectively. The ceramide composition was found to be dominated by 2-hydroxylated docosanoic (C(22h:0)), tricosanoic (C(23h:0)) and tetracosanoic (C(24h:0)) acids, and C(17) sphingosine (C(d17:1)) (where (h) is hydroxylated and (d) is dihydroxylated).

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Selected References

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  1. Baumeister S., Dennis R. D., Klünder R., Schares G., Zahner H., Geyer E. Litomosoides carinii: macrofilariae-derived glycolipids--chromatography, serology and potential in the evaluation of anthelminthic efficacy. Parasite Immunol. 1994 Dec;16(12):629–641. doi: 10.1111/j.1365-3024.1994.tb00319.x. [DOI] [PubMed] [Google Scholar]
  2. Deehan M. R., Frame M. J., Parkhouse R. M., Seatter S. D., Reid S. D., Harnett M. M., Harnett W. A phosphorylcholine-containing filarial nematode-secreted product disrupts B lymphocyte activation by targeting key proliferative signaling pathways. J Immunol. 1998 Mar 15;160(6):2692–2699. [PubMed] [Google Scholar]
  3. Deehan M. R., Harnett M. M., Harnett W. A filarial nematode secreted product differentially modulates expression and activation of protein kinase C isoforms in B lymphocytes. J Immunol. 1997 Dec 15;159(12):6105–6111. [PubMed] [Google Scholar]
  4. Dennis R. D., Baumeister S., Smuda C., Lochnit C., Waider T., Geyer E. Initiation of chemical studies on the immunoreactive glycolipids of adult Ascaris suum. Parasitology. 1995 Jun;110(Pt 5):611–623. doi: 10.1017/s0031182000065331. [DOI] [PubMed] [Google Scholar]
  5. Dennis R. D., Geyer R., Egge H., Peter-Katalinic J., Li S. C., Stirm S., Wiegandt H. Glycosphingolipids in insects. Chemical structures of ceramide tetra-, penta-, hexa-, and heptasaccharides from Calliphora vicina pupae (Insecta: Diptera). J Biol Chem. 1985 May 10;260(9):5370–5375. [PubMed] [Google Scholar]
  6. Dennis R. D., Lochnit G., Geyer R. Strategies for preliminary characterization of novel amphoteric glycosphingolipids. Methods Mol Biol. 1998;76:197–212. doi: 10.1385/0-89603-355-4:197. [DOI] [PubMed] [Google Scholar]
  7. Garate T., Kliks M. M., Cabrera Z., Parkhouse R. M. Specific and cross-reacting antibodies in human responses to Onchocerca volvulus and Dracunculus medinensis infections. Am J Trop Med Hyg. 1990 Feb;42(2):140–147. doi: 10.4269/ajtmh.1990.42.140. [DOI] [PubMed] [Google Scholar]
  8. Gerdt S., Dennis R. D., Borgonie G., Schnabel R., Geyer R. Isolation, characterization and immunolocalization of phosphorylcholine-substituted glycolipids in developmental stages of Caenorhabditis elegans. Eur J Biochem. 1999 Dec;266(3):952–963. doi: 10.1046/j.1432-1327.1999.00937.x. [DOI] [PubMed] [Google Scholar]
  9. Geyer H., Schmitt S., Wuhrer M., Geyer R. Structural analysis of glycoconjugates by on-target enzymatic digestion and MALDI-TOF-MS. Anal Chem. 1999 Jan 15;71(2):476–482. doi: 10.1021/ac980712w. [DOI] [PubMed] [Google Scholar]
  10. Geyer R., Geyer H., Kühnhardt S., Mink W., Stirm S. Capillary gas chromatography of methylhexitol acetates obtained upon methylation of N-glycosidically linked glycoprotein oligosaccharides. Anal Biochem. 1982 Apr;121(2):263–274. doi: 10.1016/0003-2697(82)90478-x. [DOI] [PubMed] [Google Scholar]
  11. Geyer R., Geyer H. Saccharide linkage analysis using methylation and other techniques. Methods Enzymol. 1994;230:86–108. doi: 10.1016/0076-6879(94)30009-7. [DOI] [PubMed] [Google Scholar]
  12. Harnett M. M., Deehan M. R., Williams D. M., Harnett W. Induction of signalling anergy via the T-cell receptor in cultured Jurkat T cells by pre-exposure to a filarial nematode secreted product. Parasite Immunol. 1998 Nov;20(11):551–563. doi: 10.1046/j.1365-3024.1998.00181.x. [DOI] [PubMed] [Google Scholar]
  13. Harnett W., Harnett M. M. Inhibition of murine B cell proliferation and down-regulation of protein kinase C levels by a phosphorylcholine-containing filarial excretory-secretory product. J Immunol. 1993 Nov 1;151(9):4829–4837. [PubMed] [Google Scholar]
  14. Harnett W., Harnett M. M. Phosphorylcholine: friend or foe of the immune system? Immunol Today. 1999 Mar;20(3):125–129. doi: 10.1016/s0167-5699(98)01419-4. [DOI] [PubMed] [Google Scholar]
  15. Harnett W., Houston K. M., Amess R., Worms M. J. Acanthocheilonema viteae: phosphorylcholine is attached to the major excretory-secretory product via an N-linked glycan. Exp Parasitol. 1993 Dec;77(4):498–502. doi: 10.1006/expr.1993.1113. [DOI] [PubMed] [Google Scholar]
  16. Harnett W., Worms M. J., Kapil A., Grainger M., Parkhouse R. M. Origin, kinetics of circulation and fate in vivo of the major excretory-secretory product of Acanthocheilonema viteae. Parasitology. 1989 Oct;99(Pt 2):229–239. doi: 10.1017/s0031182000058686. [DOI] [PubMed] [Google Scholar]
  17. Haslam S. M., Houston K. M., Harnett W., Reason A. J., Morris H. R., Dell A. Structural studies of N-glycans of filarial parasites. Conservation of phosphorylcholine-substituted glycans among species and discovery of novel chito-oligomers. J Biol Chem. 1999 Jul 23;274(30):20953–20960. doi: 10.1074/jbc.274.30.20953. [DOI] [PubMed] [Google Scholar]
  18. Haslam S. M., Khoo K. H., Houston K. M., Harnett W., Morris H. R., Dell A. Characterisation of the phosphorylcholine-containing N-linked oligosaccharides in the excretory-secretory 62 kDa glycoprotein of Acanthocheilonema viteae. Mol Biochem Parasitol. 1997 Mar;85(1):53–66. doi: 10.1016/s0166-6851(96)02807-1. [DOI] [PubMed] [Google Scholar]
  19. Helling F., Dennis R. D., Weske B., Nores G., Peter-Katalinic J., Dabrowski U., Egge H., Wiegandt H. Glycosphingolipids in insects. The amphoteric moiety, N-acetylglucosamine-linked phosphoethanolamine, distinguishes a group of ceramide oligosaccharides from the pupae of Calliphora vicina (Insecta: Diptera). Eur J Biochem. 1991 Sep 1;200(2):409–421. doi: 10.1111/j.1432-1033.1991.tb16199.x. [DOI] [PubMed] [Google Scholar]
  20. Houston K. M., Cushley W., Harnett W. Studies on the site and mechanism of attachment of phosphorylcholine to a filarial nematode secreted glycoprotein. J Biol Chem. 1997 Jan 17;272(3):1527–1533. doi: 10.1074/jbc.272.3.1527. [DOI] [PubMed] [Google Scholar]
  21. Houston K. M., Harnett W. Prevention of attachment of phosphorylcholine to a major excretory-secretory product of Acanthocheilonema viteae using tunicamycin. J Parasitol. 1996 Apr;82(2):320–324. [PubMed] [Google Scholar]
  22. Lal R. B., Kumaraswami V., Steel C., Nutman T. B. Phosphocholine-containing antigens of Brugia malayi nonspecifically suppress lymphocyte function. Am J Trop Med Hyg. 1990 Jan;42(1):56–64. doi: 10.4269/ajtmh.1990.42.56. [DOI] [PubMed] [Google Scholar]
  23. Lal R. B., Lynch T. J., Nutman T. B. Brugia malayi antigens associated with lymphocyte activation in filariasis. J Immunol. 1987 Sep 1;139(5):1652–1657. [PubMed] [Google Scholar]
  24. Lal R. B., Ottesen E. A. Phosphocholine epitopes on helminth and protozoal parasites and their presence in the circulation of infected human patients. Trans R Soc Trop Med Hyg. 1989 Sep-Oct;83(5):652–655. doi: 10.1016/0035-9203(89)90387-8. [DOI] [PubMed] [Google Scholar]
  25. Lobos E., Ondo A., Ottesen E. A., Nutman T. B. Biochemical and immunologic characterization of a major IgE-inducing filarial antigen of Brugia malayi and implications for the pathogenesis of tropical pulmonary eosinophilia. J Immunol. 1992 Nov 1;149(9):3029–3034. [PubMed] [Google Scholar]
  26. Lochnit G., Dennis R. D., Ulmer A. J., Geyer R. Structural elucidation and monokine-inducing activity of two biologically active zwitterionic glycosphingolipids derived from the porcine parasitic nematode Ascaris suum. J Biol Chem. 1998 Jan 2;273(1):466–474. doi: 10.1074/jbc.273.1.466. [DOI] [PubMed] [Google Scholar]
  27. Lochnit G., Dennis R. D., Zähringer U., Geyer R. Structural analysis of neutral glycosphingolipids from Ascaris suum adults (Nematoda:Ascaridida). Glycoconj J. 1997 Apr;14(3):389–399. doi: 10.1023/a:1018530914067. [DOI] [PubMed] [Google Scholar]
  28. Lüder C. G., Schulz-Key H., Banla M., Pritze S., Soboslay P. T. Immunoregulation in onchocerciasis: predominance of Th1-type responsiveness to low molecular weight antigens of Onchocerca volvulus in exposed individuals without microfilaridermia and clinical disease. Clin Exp Immunol. 1996 Aug;105(2):245–253. doi: 10.1046/j.1365-2249.1996.d01-747.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. MacDonald M., Copeman D. B., Harnett W. Do excretory-secretory products of Onchocerca gibsoni contain phosphorylcholine attached to O-type glycans? Int J Parasitol. 1996 Oct;26(10):1075–1080. [PubMed] [Google Scholar]
  30. Maloney M. D., Semprevivo L. H. Thin-layer and liquid column chromatographic analyses of the lipids of adult Onchocerca gibsoni. Parasitol Res. 1991;77(4):294–300. doi: 10.1007/BF00930904. [DOI] [PubMed] [Google Scholar]
  31. Sidow A., Thomas W. K. A molecular evolutionary framework for eukaryotic model organisms. Curr Biol. 1994 Jul 1;4(7):596–603. doi: 10.1016/s0960-9822(00)00131-7. [DOI] [PubMed] [Google Scholar]
  32. Soboslay P. T., Dreweck C. M., Taylor H. R., Brotman B., Wenk P., Greene B. M. Experimental onchocerciasis in chimpanzees. Cell-mediated immune responses, and production and effects of IL-1 and IL-2 with Onchocerca volvulus infection. J Immunol. 1991 Jul 1;147(1):346–353. [PubMed] [Google Scholar]
  33. Soboslay P. T., Lüder C. G., Riesch S., Geiger S. M., Banla M., Batchassi E., Stadler A., Schulz-Key H. Regulatory effects of Th1-type (IFN-gamma, IL-12) and Th2-type cytokines (IL-10, IL-13) on parasite-specific cellular responsiveness in Onchocerca volvulus-infected humans and exposed endemic controls. Immunology. 1999 Jun;97(2):219–225. doi: 10.1046/j.1365-2567.1999.00018.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wenger J. D., Forsyth K. P., Kazura J. W. Identification of phosphorylcholine epitope-containing antigens in Brugia malayi and relation of serum epitope levels to infection status of jirds with brugian filariasis. Am J Trop Med Hyg. 1988 Jan;38(1):133–141. doi: 10.4269/ajtmh.1988.38.133. [DOI] [PubMed] [Google Scholar]
  35. Weske B., Dennis R. D., Helling F., Keller M., Nores G. A., Peter-Katalinic J., Egge H., Dabrowski U., Wiegandt H. Glycosphingolipids in insects. Chemical structures of two variants of a glucuronic-acid-containing ceramide hexasaccharide from a pupae of Calliphora vicina (Insecta: Diptera), distinguished by a N-acetylglucosamine-bound phosphoethanolamine sidechain. Eur J Biochem. 1990 Jul 31;191(2):379–388. doi: 10.1111/j.1432-1033.1990.tb19133.x. [DOI] [PubMed] [Google Scholar]
  36. Wuhrer M., Dennis R. D., Doenhoff M. J., Bickle Q., Lochnit G., Geyer R. Immunochemical characterisation of Schistosoma mansoni glycolipid antigens. Mol Biochem Parasitol. 1999 Oct 15;103(2):155–169. doi: 10.1016/s0166-6851(99)00123-1. [DOI] [PubMed] [Google Scholar]
  37. Zanetta J. P., Timmerman P., Leroy Y. Gas-liquid chromatography of the heptafluorobutyrate derivatives of the O-methyl-glycosides on capillary columns: a method for the quantitative determination of the monosaccharide composition of glycoproteins and glycolipids. Glycobiology. 1999 Mar;9(3):255–266. doi: 10.1093/glycob/9.3.255. [DOI] [PubMed] [Google Scholar]

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