Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1994 Dec;62(12):5424–5433. doi: 10.1128/iai.62.12.5424-5433.1994

Chemical and immunological characterization of the extracellular galactomannan of Aspergillus fumigatus.

J P Latgé 1, H Kobayashi 1, J P Debeaupuis 1, M Diaquin 1, J Sarfati 1, J M Wieruszeski 1, E Parra 1, J P Bouchara 1, B Fournet 1
PMCID: PMC303284  PMID: 7960122

Abstract

The galactomannan (GM) produced extracellularly by Aspergillus fumigatus has been purified by a double sequential hydrazine-nitrous acid treatment of the ethanol precipitate of the culture filtrate. Nuclear magnetic resonance and gas-liquid chromatography-mass spectrometry analysis have been performed on intact GM, acid-hydrolyzed GM, and oligomers resulting from the acetolysis of the acid-hydrolyzed GM. Results show that A. fumigatus GM is composed of a linear mannan core with an alpha-(1-2)-linked mannotetraose repeating unit attached via alpha-(1-6) linkage. Side chains composed of an average of 4 to 5 beta-(1-5)-galactofuranose units are linked to C-6 and C-3 positions of alpha-(1-2)-linked mannose units of the mannan. The immunoreactivity of GM and HCl-hydrolyzed GM was studied by use of human sera from aspergillosis patients and an antigalactofuran monoclonal antibody. The alpha-(1-2) (1-6)-mannan core is not antigenic. The immunogenic galactofuran is found amongst several exocellular glycoproteins. According to a direct enzyme-linked immunosorbent assay with GM as the detector antigen, only 26% of the serum samples from aspergilloma patients (all positive by immunodiffusion assays) give optical density values superior to a cutoff estimated as the mean +/- 3 standard deviations of values obtained with control sera.

Full text

PDF
5424

Images in this article

5430Image
on p.5430

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. AMES B. N., DUBIN D. T. The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid. J Biol Chem. 1960 Mar;235:769–775. [PubMed] [Google Scholar]
  2. Azuma I., Kimura H., Hirao F., Tsubura E., Yamamura Y., Misaki A. Biochemical and immunological studies on Aspergillus. 3. Chemical and immunological properties of glycopeptide obtained from Aspergillus fumigatus. Jpn J Microbiol. 1971 May;15(3):237–246. [PubMed] [Google Scholar]
  3. Bardalaye P. C., Nordin J. H. Chemical structure of the galactomannan from the cell wall of Aspergillus niger. J Biol Chem. 1977 Apr 25;252(8):2584–2591. [PubMed] [Google Scholar]
  4. Bennett J. E., Bhattacharjee A. K., Glaudemans C. P. Galactofuranosyl groups are immunodominant in Aspergillus fumigatus galactomannan. Mol Immunol. 1985 Mar;22(3):251–254. doi: 10.1016/0161-5890(85)90158-0. [DOI] [PubMed] [Google Scholar]
  5. Colson P., Jennings H. J., Smith I. C. Composition, sequence, and conformation of polymers and oligomers of glucose as revealed by carbon-13 nuclear magentic resonance. J Am Chem Soc. 1974 Dec 25;96(26):8081–8087. doi: 10.1021/ja00833a038. [DOI] [PubMed] [Google Scholar]
  6. Datema R., Wessels J. G., van den Ende H. The hyphal wall of Mucor mucedo. 2. Hexosamine-containing polymers. Eur J Biochem. 1977 Nov 1;80(2):621–626. doi: 10.1111/j.1432-1033.1977.tb11919.x. [DOI] [PubMed] [Google Scholar]
  7. Dupont B., Huber M., Kim S. J., Bennett J. E. Galactomannan antigenemia and antigenuria in aspergillosis: studies in patients and experimentally infected rabbits. J Infect Dis. 1987 Jan;155(1):1–11. doi: 10.1093/infdis/155.1.1. [DOI] [PubMed] [Google Scholar]
  8. Fournet B., Strecker G., Leroy Y., Montreuil J. Gas--liquid chromatography and mass spectrometry of methylated and acetylated methyl glycosides. Application to the structural analysis of glycoprotein glycans. Anal Biochem. 1981 Sep 15;116(2):489–502. doi: 10.1016/0003-2697(81)90393-6. [DOI] [PubMed] [Google Scholar]
  9. Gander J. E. Fungal cell wall glycoproteins and peptido-polysaccharides. Annu Rev Microbiol. 1974;28(0):103–119. doi: 10.1146/annurev.mi.28.100174.000535. [DOI] [PubMed] [Google Scholar]
  10. Gander J. E., Jentoft N. H., Drewes L. R., Rick P. D. The 5-O-beta-D-galactofuranosyl-containing exocellular glycopeptide of Penicillium charlesii. Characterization of the phosphogalactomannan. J Biol Chem. 1974 Apr 10;249(7):2063–2072. [PubMed] [Google Scholar]
  11. Hearn V. M., Griffiths B. L., Gorin P. A. Structural analysis of water-soluble fractions obtained from Aspergillus fumigatus mycelium. Glycoconj J. 1989;6(1):85–100. doi: 10.1007/BF01047892. [DOI] [PubMed] [Google Scholar]
  12. Kamerling J. P., Gerwig G. J., Vliegenthart J. F., Clamp J. R. Characterization by gas-liquid chromatography-mass spectrometry and proton-magnetic-resonance spectroscopy of pertrimethylsilyl methyl glycosides obtained in the methanolysis of glycoproteins and glycopeptides. Biochem J. 1975 Dec;151(3):491–495. doi: 10.1042/bj1510491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kobayashi H., Debeaupuis J. P., Bouchara J. P., Latge J. P. An 88-kilodalton antigen secreted by Aspergillus fumigatus. Infect Immun. 1993 Nov;61(11):4767–4771. doi: 10.1128/iai.61.11.4767-4771.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kobayashi H., Shibata N., Mitobe H., Ohkubo Y., Suzuki S. Structural study of phosphomannan of yeast-form cells of Candida albicans J-1012 strain with special reference to application of mild acetolysis. Arch Biochem Biophys. 1989 Aug 1;272(2):364–375. doi: 10.1016/0003-9861(89)90230-0. [DOI] [PubMed] [Google Scholar]
  15. Kobayashi H., Shibata N., Nakada M., Chaki S., Mizugami K., Ohkubo Y., Suzuki S. Structural study of cell wall phosphomannan of Candida albicans NIH B-792 (serotype B) strain, with special reference to 1H and 13C NMR analyses of acid-labile oligomannosyl residues. Arch Biochem Biophys. 1990 Apr;278(1):195–204. doi: 10.1016/0003-9861(90)90248-w. [DOI] [PubMed] [Google Scholar]
  16. Kobayashi H., Shibata N., Suzuki S. Acetolysis of Pichia pastoris IFO 0948 strain mannan containing alpha-1,2 and beta-1,2 linkages using acetolysis medium of low sulfuric acid concentration. Arch Biochem Biophys. 1986 Mar;245(2):494–503. doi: 10.1016/0003-9861(86)90242-0. [DOI] [PubMed] [Google Scholar]
  17. Kocourek J., Ballou C. E. Method for fingerprinting yeast cell wall mannans. J Bacteriol. 1969 Dec;100(3):1175–1181. doi: 10.1128/jb.100.3.1175-1181.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Latgé J. P., Moutaouakil M., Debeaupuis J. P., Bouchara J. P., Haynes K., Prévost M. C. The 18-kilodalton antigen secreted by Aspergillus fumigatus. Infect Immun. 1991 Aug;59(8):2586–2594. doi: 10.1128/iai.59.8.2586-2594.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lloyd K. O. Isolation, characterization, and partial structure of peptido galactomannans from the yeast form of Cladosporium werneckii. Biochemistry. 1970 Aug 18;9(17):3446–3453. doi: 10.1021/bi00819a025. [DOI] [PubMed] [Google Scholar]
  20. Notermans S., Veeneman G. H., van Zuylen C. W., Hoogerhout P., van Boom J. H. (1----5)-linked beta-D-galactofuranosides are immunodominant in extracellular polysaccharides of Penicillium and Aspergillus species. Mol Immunol. 1988 Oct;25(10):975–979. doi: 10.1016/0161-5890(88)90003-x. [DOI] [PubMed] [Google Scholar]
  21. Parente J. P., Cardon P., Leroy Y., Montreuil J., Fournet B., Ricart G. A convenient method for methylation of glycoprotein glycans in small amounts by using lithium methylsulfinyl carbanion. Carbohydr Res. 1985 Aug 15;141(1):41–47. doi: 10.1016/s0008-6215(00)90753-5. [DOI] [PubMed] [Google Scholar]
  22. Parra E., Jiménez-Barbero J., Bernabé M., Leal J. A., Prieto A., Gómez-Miranda B. Structural studies of fungal cell-wall polysaccharides from two strains of Talaromyces flavus. Carbohydr Res. 1994 Jan 3;251:315–325. doi: 10.1016/0008-6215(94)84294-9. [DOI] [PubMed] [Google Scholar]
  23. Reiss E., Lehmann P. F. Galactomannan antigenemia in invasive aspergillosis. Infect Immun. 1979 Jul;25(1):357–365. doi: 10.1128/iai.25.1.357-365.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rick P. D., Drewes L. R., Gander J. E. The 5-O-beta-D-galactofuranosyl-containing exocellular glycopeptide of Penicillium charlesii. Occurrence of ethanolamine and partial characterization of the peptide portion and the carbohydrate-peptide linkage. J Biol Chem. 1974 Apr 10;249(7):2073–2078. [PubMed] [Google Scholar]
  25. Rietschel-Berst M., Jentoft N. H., Rick P. D., Pletcher C., Fang F., Gander J. E. Extracellular exo-beta-galactofuranosidase from Penicillium charlesii: isolation, purification, and properties. J Biol Chem. 1977 May 25;252(10):3219–3226. [PubMed] [Google Scholar]
  26. Rogers T. R., Haynes K. A., Barnes R. A. Value of antigen detection in predicting invasive pulmonary aspergillosis. Lancet. 1990 Nov 17;336(8725):1210–1213. doi: 10.1016/0140-6736(90)92831-2. [DOI] [PubMed] [Google Scholar]
  27. Sakaguchi O., Yokota K., Suzuki M. Immunochemical and biochemical studies of fungi. 13. On the galactomannans isolated from mycelia and culture filtrates of several filamentous fungi. Jpn J Microbiol. 1969 Mar;13(1):1–7. [PubMed] [Google Scholar]
  28. Stynen D., Sarfati J., Goris A., Prévost M. C., Lesourd M., Kamphuis H., Darras V., Latgé J. P. Rat monoclonal antibodies against Aspergillus galactomannan. Infect Immun. 1992 Jun;60(6):2237–2245. doi: 10.1128/iai.60.6.2237-2245.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Unkefer C. J., Gander J. E. The 5-O-beta-D-galactofuranosyl-containing glycopeptide from Penicillium charlesii. Carbon 13 nuclear magnetic resonance studies. J Biol Chem. 1979 Dec 10;254(23):12131–12135. [PubMed] [Google Scholar]
  30. Unkefer C. J., Gander J. E. The 5-O-beta-D-galactofuranosyl-containing peptidophosphogalactomannan of Penicillium charlesii. Characterization of the mannan by 13C NMR spectroscopy. J Biol Chem. 1990 Jan 15;265(2):685–689. [PubMed] [Google Scholar]
  31. Unkefer C. J., Jackson C., Gander J. E. The 5-O-beta-D-galactofuranosyl-containing glycopeptide from Penicillium charlesii. Identification of phosphocholine attached to C-6 of mannopyranosyl residues of the mannan region. J Biol Chem. 1982 Mar 10;257(5):2491–2497. [PubMed] [Google Scholar]
  32. Van Bruggen-Van Der Lugt A. W., Kamphuis H. J., De Ruiter G. A., Mischnick P., Van Boom J. H., Rombouts F. M. New structural features of the antigenic extracellular polysaccharides of Penicillium and Aspergillus species revealed with exo-beta-D-galactofuranosidase. J Bacteriol. 1992 Oct;174(19):6096–6102. doi: 10.1128/jb.174.19.6096-6102.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Zanetta J. P., Breckenridge W. C., Vincendon G. Analysis of monosaccharides by gas-liquid chromatography of the O-methyl glycosides as trifluoroacetate derivatives. Application to glycoproteins and glycolipids. J Chromatogr. 1972 Jul 5;69(2):291–304. doi: 10.1016/s0021-9673(00)92897-8. [DOI] [PubMed] [Google Scholar]
  34. de Repentigny L., Boushira M., Ste-Marie L., Bosisio G. Detection of galactomannan antigenemia by enzyme immunoassay in experimental invasive aspergillosis. J Clin Microbiol. 1987 May;25(5):863–867. doi: 10.1128/jcm.25.5.863-867.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES