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Glycoconjugate Journal

, Volume 23, Issue 7–8, pp 463–471 | Cite as

Galactosylation of IgG from rheumatoid arthritis (RA) patients – changes during therapy

  • Marta Pasek
  • Maria Duk
  • Maria Podbielska
  • Renata Sokolik
  • Jacek Szechiński
  • Elwira Lisowska
  • Hubert KrotkiewskiEmail author
Article

Abstract

It is well documented that serum IgG from rheumatoid arthritis (RA) patients exhibits decreased galactosylation of its conservative N-glycans (Asn-297) in CH2 domains of the heavy chains; it has been shown that this agalactosylation is proportional to disease severity. In the present investigation we analyzed galactosylation of IgG derived from the patients using a modified ELISA-plate test, biosensor BIAcore and total sugar analysis (GC-MS). For ELISA and BIAcore the binding of IgG preparations, purified from the patients’ sera, to two lectins: Ricinus communis (RCA-I) and Griffonia simplicifolia (GSL-II) was applied. Based on ELISA-plate test an agalactosylation factor (AF, a relative ratio of GSL-II/RCA-I binding) was calculated, which was proportional to actual disease severity. Repeated testing of several patients before and after treatment with methotrexate (MTX) alone or in combination with Remicade (a chimeric antibody anti-TNF-α) supplied results indicating an increase of IgG galactosylation during the treatment. This introductory observation suggests that IgG galactosylation may be an additional indicator of the RA patients’ improvement.

Keywords

IgG Rheumatoid arthritis Surface plasmon resonance Ricinus communis lectin Griffonia simplicifolia lectin 

Abbreviations

IgG

immunoglobulin G

RA

rheumatoid arthritis

RCA

lectin from Ricinus communis

PVL

lectin from Psathyrella velutina

GSL

lectin from Griffonia simplicifolia

ESR

erythrocyte sedimentation rate

CRP

C-reactive protein

GC-MS

gas chromatography-mass spectrometry

MTX

methotrexate

NC

nitrocellulose

DTT

dithiotreitol

AF

agalactosylation factor

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References

  1. 1.
    Lee, D.M., Weinblatt, M.E.: Rheumatoid arthritis. The Lancet 358, 903–11 (2001)CrossRefGoogle Scholar
  2. 2.
    Wormald, M.R., Rudd, P.M., Harvey, D.J., Chang, S.C., Scragg, I.G., Dwek, R.A.: Variations of oligosaccharide-protein interactions in immunoglobulin G determine the site-specific glycosylation profiles and modulate the dynamic motion of the Fc oligosaccharides. Biochem. 36, 1370–180 (1997)CrossRefGoogle Scholar
  3. 3.
    Lisowska, E., Duk, M., Wu, A.: Preparation of biotinylated lectins and application in microtiter plate assays and western blotting. BioMethods 7, 115–29 (1996)Google Scholar
  4. 4.
    Arnett, F.C., Edworthy, S.M., Bloch, D.A., McShane, D.J., Freies, J.F., Cooper, N.S., Healey, L.A., Kaplan, S.R., Liang, M.H., Luthra, H.S.: The American Rheumatism Association 1987 revised criteria for classification of rheumatoid arthritis. Arthritis Rheum 31, 315–24 (1988)PubMedGoogle Scholar
  5. 5.
    Ey, P.L., Prowse, S.J., Jenkin, C.R.: Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-Sepharose. Immunochemistry 15, 429–36 (1978)PubMedCrossRefGoogle Scholar
  6. 6.
    Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–85 (1970)PubMedCrossRefGoogle Scholar
  7. 7.
    Schaffner, W., Weissmann, C.: A rapid, sensitive and specific method for the determination of protein in dilute solution, Anal. Biochem. 56, 502–14 (1973)Google Scholar
  8. 8.
    Liljeblad, M., Lundblad, A., Påhlsson, P.: Analysis of agalacto-IgG in rheumatoid arthritis using surface plasmon resonance. Glycoconjugate J. 17, 323–29 (2000)CrossRefGoogle Scholar
  9. 9.
    Sawardeker, J.S., Sloneker, J.H., Jeanes, A.: Quantitative determination of monosaccharides as their alditol acetates by liquid chromatography. Anal. Chem. 37, 1602–4 (1965)CrossRefGoogle Scholar
  10. 10.
    Towbin, H., Stachelin, T., Gordon, J.: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some application. Proc. Natl. Acad. Sci. USA. 76, 4350–54 (1979)PubMedCrossRefGoogle Scholar
  11. 11.
    Krotkiewska, B., Pasek, M., Krotkiewski, H.: Interaction of glycophorin A with lectins as measured by surface plasmon resonance (SPR). Acta. Biochim. Pol. 49, 481–90 (2002)PubMedGoogle Scholar
  12. 12.
    Lin, T.T.S., Li, S.S.L.: Purification and physicochemical properties of ricins and agglutinins from, Ricinus communis. Eur. J. Biochem. 105, 453–59 (1980)Google Scholar
  13. 13.
    Ebisu, S., Iyer, P.N.S., Goldstein, I.J.: Equlibrium dialysis and carbohydrate-binding studies on the 2-acetamido-2-deoxy-D-glucopyranosyl-binding lectin from Bandeiraea simplicifolia seeds, Carbohydr. Res. 61, 129–38 (1978)Google Scholar
  14. 14.
    Ueda, H., Matsumoto, H., Takahashi, N., Ogawa, H.: Psathyrella velutina mushroom lectin exhibits high affinity toward sialoglycoproteins possessing terminal N-acetylneuraminic acid a2,3-linked to penultimate galactose residues of trisialyl N-glycans. J. Biol. Chem. 277, 24916–25 (2002)PubMedCrossRefGoogle Scholar
  15. 15.
    Tomana, M., Schrohenloher, R.E., Koopman, W.J., Alarcon, G.S., Paul, W.A.: Abnormal glycosylation of serum IgG from patients with chronic inflammatory diseases. Arthritis Rheum. 31, 333–8 (1988)PubMedGoogle Scholar
  16. 16.
    Parekh, R.B., Dwek, R.A., Sutton, B.J., Fernandes, D.L., Leung, A., Stanworth, D., Rademacher, T.W., Mizuochi, T., Taniguchi, T., Matsuta, K., Takeuchi, F., Nagano, Y., Miyamoto, T., Kobata, A.: Association of rheumatoid arthtritis and primary osteoarthritis with changes in the glycosylation pattern of total serum IgG. Nature 316, 452–7 (1985)PubMedCrossRefGoogle Scholar
  17. 17.
    Takahashi, N., Ishii, I., Ishihara, H., Masami, M., Tejima, S., Jefferis, R., Endo, S., Arata, Y.: Comparative structural study of the N-linked oligosaccharides of human normal and pathological immunoglobulin G. Biochem. 26, 1137–44 (1987)CrossRefGoogle Scholar
  18. 18.
    Watson, M., Rudd, P.M., Bland, M., Dwek, R.A., Axford, J.S.: Sugar printing rheumatic diseases. A potential method for disease differentiation using immunoglobulin G oligosaccharides. Arthritis Rheum 42, 1682–90 (1999)PubMedCrossRefGoogle Scholar
  19. 19.
    Harada, H., Kamei, M., Tokumoto, Y., Yui, S., Koyama, F., Kochibe, N., Endo, T., Kobata, A.: Systematic fractionation of oligosaccharides of human immunoglobulin G by serial affinity chromatography on immobilized lectin columns. Anal. Biochem. 164, 374–81 (1987)PubMedCrossRefGoogle Scholar
  20. 20.
    Sumar, N., Bodman, K.B., Rademacher, T.W., Dwek, R.A., Williams, P., Parekh, R.B., Edge, J., Rook, G.A.W., Isenberg, D.A., Hay, F.C., Roitt, I.M.: Analysis of glycosylation changes in IgG using lectins. J. Immunol. Meth. 131, 127–36 (1990)CrossRefGoogle Scholar
  21. 21.
    Bond, A., Jones, M.G., Hay, F.C.: Human IgG preparations isolated by ion-exchange chromatography differ in their glycosylation profiles. J. Immunol. Meth. 166, 27–33 (1993)CrossRefGoogle Scholar
  22. 22.
    Flögel, M., Lauc, G., Gornik, I., Maček, B.: Fucosylation and galactosylation of IgG heavy chains differ between acute and remission phases of juvenile chronic arthritis. Clin. Chem. Lab. Med. 36, 99–102 (1998)PubMedCrossRefGoogle Scholar
  23. 23.
    Tsuchiya, N., Endo, T., Matsuta, K., Yoshinoya, S., Takeuchi, F., Nagano, Y., Shiota, M., Furukawa, K., Kochibe, N., Ito, K., Kobata, A.: Detection of glycosylation abnormality in rheumatoid IgG using N-acetylglucosamine-specific Psathyrella velutina lectin. J. Immunol. 151, 1137–46 (1993)PubMedGoogle Scholar
  24. 24.
    Rahman, A., Isenberg, D.: Does it take sugar? A clinical role for measuring the glycosylation of IgG. Ann. Rheum. Dis. 54, 689–91 (1995)PubMedCrossRefGoogle Scholar
  25. 25.
    Szechiński, J., Wiland, P., Szczepański, L., Chudzik, D., Sierakowski, S., Klimiuk, P.A.: One year combined treatment with methotrexate and monoclonal antibody anti-TNFα (Remicade) in patients with active rheumatoid arthritis–multicenter clinical trial. Reumatologia (Polish) 39, 200–7 (2001)Google Scholar

Copyright information

© Springer Science + Business Media, LLC 2006

Authors and Affiliations

  • Marta Pasek
    • 1
  • Maria Duk
    • 1
  • Maria Podbielska
    • 1
  • Renata Sokolik
    • 2
  • Jacek Szechiński
    • 2
  • Elwira Lisowska
    • 1
  • Hubert Krotkiewski
    • 1
    Email author
  1. 1.Ludwik Hirszfeld Institute of Immunology & Experimental TherapyPolish Academy of SciencesWroclawPoland
  2. 2.Department of RheumatologyMedical UniversityWroclawPoland

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